Technical assessment of patterning methods.
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More than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\\n\\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\\n\\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\\n\\nAdditionally, each book published by IntechOpen contains original content and research findings.
\\n\\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:{caption:"IntechOpen Maintains",originalUrl:"/media/original/113"}},components:[{type:"htmlEditorComponent",content:'
Simba Information has released its Open Access Book Publishing 2020 - 2024 report and has again identified IntechOpen as the world’s largest Open Access book publisher by title count.
\n\nSimba Information is a leading provider for market intelligence and forecasts in the media and publishing industry. The report, published every year, provides an overview and financial outlook for the global professional e-book publishing market.
\n\nIntechOpen, De Gruyter, and Frontiers are the largest OA book publishers by title count, with IntechOpen coming in at first place with 5,101 OA books published, a good 1,782 titles ahead of the nearest competitor.
\n\nSince the first Open Access Book Publishing report published in 2016, IntechOpen has held the top stop each year.
\n\n\n\nMore than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\n\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\n\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\n\nAdditionally, each book published by IntechOpen contains original content and research findings.
\n\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\n\n\n\n
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Ghofrani",coverURL:"https://cdn.intechopen.com/books/images_new/10597.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"183482",title:"Dr.",name:"Mahmoud",middleName:null,surname:"Ghofrani",slug:"mahmoud-ghofrani",fullName:"Mahmoud Ghofrani"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}}},ofsBook:{item:{type:"book",id:"11467",leadTitle:null,title:"Bismuth-Based Nanostructured Materials",subtitle:null,reviewType:"peer-reviewed",abstract:"
\r\n\tBismuth-based nanostructured materials have received increasing research interest in the past decades, especially for their applications in photocatalysis and electrocatalysis. New bismuth-based nanostructured materials have been fabricated, and their optical and electronic structures can be fine-tuned via various synthetic approaches. These bismuth-based materials have been widely applied in photocatalysis (NOx removal, VOCs purification, CO2 reduction, water splitting, organic pollutants degradation, heavy metals reduction) and electrocatalysis (nitrogen fixation, CO2 reduction, water electrolysis, organic synthesis). The rapid development in this field needs a comprehensive summary to reflect the new advances in recent years. The aim of this project is to invite researchers worldwide to contribute to this field and promote the developments in the synthesis, characterization, structure-property relationship determination, and application of bismuth-based catalysts, proposing organized materials, challenges, and prospects to guide future works. The content of this book could attract broad interest from diverse fields of materials, catalysis, chemistry, environment, medicine, energy, and engineering.
",isbn:"978-1-83768-048-1",printIsbn:"978-1-83768-047-4",pdfIsbn:"978-1-83768-049-8",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,isNomenclature:!1,hash:"951c872d9d90e13cfe7d97c0af91845e",bookSignature:"Dr. William Wilson Anku",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11467.jpg",keywords:"Semiconductor, Synthesis, Morphology, Shape Control, Metal Doping, Surface Modification, Catalysis, Photocatalysis, Photoelectrochemical, Nitrogen Fixation, Energy Conversion, Environmental Remediation",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"May 12th 2022",dateEndSecondStepPublish:"July 13th 2022",dateEndThirdStepPublish:"September 11th 2022",dateEndFourthStepPublish:"November 30th 2022",dateEndFifthStepPublish:"January 29th 2023",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"a month",secondStepPassed:!0,areRegistrationsClosed:!1,currentStepOfPublishingProcess:3,editedByType:null,kuFlag:!1,biosketch:"Dr. William Wilson Anku is a Research Scientist at CSIR- Water Research Institute, Accra-Ghana. He has co-authored 37 papers in renowned peer-reviewed scientific publications with over 590 citations resulting in an H-index of 12.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"196465",title:"Dr.",name:"William Wilson",middleName:null,surname:"Anku",slug:"william-wilson-anku",fullName:"William Wilson Anku",profilePictureURL:"https://mts.intechopen.com/storage/users/196465/images/system/196465.jpg",biography:'Curriculum Vitae\n of \nDr William Wilson Anku\n________________________________________\nCSIR- Water Research Institute,\nP. O. Box AH 38, \nAchimota-Accra, Ghana\n\nPrimary Email Address: williamanku85@gmail.com \nAlternate Email Address: williamanku@csir.org.gh \nMobile Numbers: +233547507987/+233577035326 \nGoogle Scholar: https://scholar.google.com/citations?user=tK_Q8UQAAAAJ&hl=en\nORCID: https://orcid.org/0000-0002-5551-6130\nResearchGate: https://www.researchgate.net/profile/William_Wilson_Anku/research\n\nPersonal Information:\nSurname: Anku\nFirst Names: William Wilson\nGender: Male\nCitizenship: Ghanaian\nDate of birth: 20/12/1976\n\nResearch Interests:\n1. Design of nanoparticles with unique structural and physical properties, and the assessment of their structure-property relationships. \n2. Development and evaluation of photocatalytic, ion exchange, adsorption/filtration properties of metal oxide semiconductors and agro-industrial wastes-based nanomaterials for their practical application in water/wastewater treatment \n3. Water/wastewater treatment\t\n\nEducation:\n2015 – 2018: PhD Chemistry, University of Johannesburg, South Africa.\n2005 – 2008: MSc Environmental Science, Kwame Nkrumah University of Science &\n Technology, Ghana.\n1999 – 2003: BSc Chemistry, Kwame Nkrumah University of Science & Technology, Ghana\n1995 – 1997: Secondary School Certificate, Saint Augustine’s College, Cape Coast, Ghana. \n\nEmployment History:\n1.\tResearch Scientist (March 2019-present): CSIR-Water Research Institute, \nAccra-Ghana.\n2.\tPostdoctoral Research Fellow (February 2018 – January 2019): Department of Applied Chemistry, University of Johannesburg, South Africa \n3.\tTeaching Assistant/Tutor (June 2015 – November 2016): Department of Applied Chemistry, University of Johannesburg, South Africa \n4.\tChemistry Tutor (January 2007-September 2014): Effiduase Senior High School, Effiduase-Ashanti (Ghana Education Service).\n\nSupervision of junior researchers at the graduate and postgraduate level:\n1.\tPhD thesis supervision:\n(a) Student Name: Michael Kumi\nInstitution: Department of Applied Chemistry, University of Johannesburg (UJ), South Africa.\nThesis title: Integrated bone and biochar bed for contaminant removal from groundwater. (In progress).\n\n(b) Student Name: George Atongo Atia\n Institution: Department of chemistry, KNUST, Kumasi\n Thesis title: Fabrication of CNTs-metal oxide/polymer chemical sensors for gas sensor\n application and computational studies. (In progress).\n\n2.\tMSc Thesis supervision:\n(a) Student Name: Esther Acheampong \nInstitution: Department of Chemical Engineering, KNUST, Kumasi\n Thesis title: Synthesis of polysulphide intercalated layered double hydroxides for\n adsorption processes. (Completed).\n\n(b)\tStudent Name: Sechaba Menyadi\nInstitution: Department of Applied Chemistry, UJ, South Africa.\nThesis title: Improving the thermoelectric performance of zinc oxide with Al3+, In3+ \nand 2D materials through the formation of superlattice structures. (Completed).\n\n(c)\tStudent Name: Nokuthula Ndaba\nInstitution: Department of Applied Chemistry, UJ, South Africa.\n Thesis title: Isolation and characterization of Drimia delagoensis phytochemicals and\n their application in diabetic foot ulcer treatment. (Completed).\n\nExternal examination of PhD/MSc theses and proposal reviews:\n1.\tExternal examination of a PhD thesis from the Chemical Engineering Department of Vaal University of Technology, South Africa, 2021.\n2.\tExternal examination of PhD thesis from the Physics and Chemistry Departments of Kwame Nkrumah University of Science and Technology, KNUST-Ghana, 2020.\n3.\tReviewer for the 2021-2022 Graduate Women in Science (GWIS) National Fellowship Program of the USA.\n\nScientific Reviewing Activities:\nServing as a reviewer for the following journals:\nACS Applied Materials and Interfaces, ACS Applied Nanomaterials, ACS Industrial and Engineering Chemistry Research, Journal of Dispersion Science and Technology, Journal of Taiwan Institute of Chemical Engineers, Electroanalysis, Journal of Molecular Catalysis A, Inorganic and Nano-Metal Chemistry, Materials Science in Semiconductor Processing, Recent Innovations in Chemical Engineering, Journal of Inorganic and Organometallic Polymers and Materials, International Journal of Biological Macromolecules, Fibers and Polymers, Catalysis letters, Desalination and water treatment, Inorganic and Nano-Metal Chemistry.\n\nProfessional Associations:\n•\tResearch Scientist Association of Ghana\n•\tSouth African Chemical Institute (SACI)\n\nLeadership and volunteering activities:\n•\tVice President of Water Research Institute Branch of Research Staff Association (RSA) of the Council for Scientific and Industrial Research (CSIR), Ghana (2021-2022).\n•\tEditorial board member of RSA-CSIR, Southern Zone (2021/2022).\n•\tEnvironmental Science Department Representative of Graduate Students Association of Ghana: Kwame Nkrumah University of Science and Technology (KNUST) branch (2005-2006).\n•\tVice President of Volta Region Students Association: KNUST branch (2002-2003).\n•\tNational Public Relations Officer of Ghana Students Chemical Society: KNUST branch (2002-2003).\n•\tGeneral Secretary of Volta Region Students Association: KNUST branch (2001-2002).\n•\tVolunteer Teacher at Asukawkaw Senior High School in the Volta Region of Ghana (May-August 2002)\n\nPrizes, awards, fellowships:\n•\tPostdoctoral research fellowship: Faculty of Science, University of Johannesburg, 2018.\n•\tPhD studentship: Faculty of Science, University of Johannesburg, 2015-2018.\n•\tStudents travel fund award: National Research Foundation (NRF) of South Africa, 2016.\n•\tBest poster presenter at the 5th UJ Cross Faculty Symposium held at UJ-Bunting Road Campus, South Africa on 13th October 2015.\n•\tSecond best poster presenter at the 3rd conference on “Emerging Frontiers for Sustainable Water” held at the Protea Hotel Wanderers, in Johannesburg, South Africa from 3-5 August 2015. \n\nPublication Record:\nA.\tBook Chapters\n1.\tOtun, Kabir Opeyemi, Idris Olayiwola Azeez, Onoyivwe Monday Ama, William Wilson Anku, Uyiosa Osagie Aigbe, Kingsley Eghonghon Ukhurebor, and Robert Birundu Onyancha. "Sensing the Presence of Inorganic Ions in Water: The Use of Electrochemical Sensors." In Modified Nanomaterials for Environmental Applications, pp. 65-89. Springer, Cham, 2022.\n2.\tAnku, William Wilson, Onoyivwe Monday Ama, Ikenna Chibuzor Emeji, Uyiosa Osagie Aigbe, Adelaja Otolorin Osibote, Peter Ogbemudia Osifo, and Suprakas Sinha Ray. “Functionalized nanomagnetic materials for environmental applications”. In Functionalized Nanomaterials Based Devices for Environmental Applications, pp. 127-145. Elsevier, 2021.\n3.\tKhoele, Khotso, Onoyivwe Monday Ama, Ikenna Chibuzor Emeji, William Wilson Anku, Suprakas Sinha Ray, David Jacobus Delport, and Peter Ogbemudia Osifo. “Dynamic Degradation Efficiency of Major Organic Pollutants from Wastewater”. Springer, Cham, In book: Nanostructured Metal-Oxide Electrode Materials for Water Purification, pp. 1-18, 2020.\n4.\tAnku, William Wilson, Onoyivwe Monday Ama, Suprakas Sinha Ray, and Peter Ogbemudia Osifo. “Application of Modified Metal Oxide Electrodes in Photoelectrochemical Removal of Organic Pollutants from Wastewater”. Springer, Cham. In book: Nanostructured Metal-Oxide Electrode Materials for Water Purification, pp. 151-166, 2020.\n5.\tWilliam W Anku, Ephraim M Kiarii, Sudheesh K Shukla, and Penny P Govender. “Photocatalytic degradation of pharmaceuticals using graphene based materials”. Springer, Cham. In book: A New Generation Material Graphene: Applications in Water Technology. pp 187-208, 2018.\n6.\tWilliam W Anku, Samuel OB Oppong and Penny P Govender. “Bismuth-based nanoparticles as photocatalytic materials”. InTechOpen. In book: Bismuth: Advanced Applications and Defects Characterization. pp 25-44, 2018.\n7.\tWilliam W Anku, Messai A Mamo and Penny P Govender. “Phenolic compounds in water: sources, reactivity, toxicity and treatment methods”. InTechOpen. In book: Phenolic Compounds-Natural Sources, Importance and Applications. pp. 420-443, 2017. \n\nB.\tPeer-Reviewed Journal Publications \n\n1. Ahiahonu, Elvis K., William W. Anku, Ashira Roopnarain, Ezekiel Green, Penny P. Govender, and Mahloro H. Serepa‐Dlamini. Bioresource potential of Tetradesmus obliquus UJEA_AD: critical evaluation of biosequestration rate, biochemical and fatty acid composition in BG11 media. Journal of Chemical Technology & Biotechnology (2021).\n2. Ahiahonu, Elvis Kodzo, William Wilson Anku, Ashira Roopnarain, Ezekiel Green, Penny Poomani Govender, and Mahloro Hope Serepa-Dlamini. Bioprospecting wild South African microalgae as a potential third-generation biofuel feedstock, biological carbon-capture agent and for nutraceutical applications. Biomass Conversion and Biorefinery (2021): 1-16.\n3. Obiri, Samuel, Gloria Addico, Saada Mohammed, Wilson William Anku, Humphry Darko, and Okrah Collins. Water quality assessment of the Tano Basin in Ghana: a multivariate statistical approach. Applied Water Science 11 (2021): 1-8.\n4. Oppong, Samuel Osei-Bonsu, Francis Opoku, William Wilson Anku, and Penny P. Govender. Insights into the complementary behaviour of Gd doping in GO/Gd/ZnO composites as an efficient candidate towards photocatalytic degradation of indigo carmine dye. Journal of Materials Science 56 (2021): 8511-8527.\n5. Ama Onoyivwe Monday, Khotso Khoele, William Wilson Anku, Suprakas Sinha Ray, Peter Ogbemudia Osifo, and David Jacobus Delport. Synthesis and Application of MnO2/Exfoliated Graphite Electrodes for Enhanced Photoelectrochemical Degradation of Methylene Blue and Congo Red Dyes in Water. Electrocatalysis.11 (2020): 413-421.\n6. Anku, William Wilson, Eric Selorm Agorku, Samuel Osei-Bonsu Oppong, and Anthony Yaw Karikari. "MWCNTs attached neodymium doped-ZnO photocatalysts for efficient removal of dyes from wastewater. SN Applied Sciences. 5 (2020): 1-13.\n7. Karikari Anthony Yaw, Asmah Ruby, Anku, William Wilson, Amisah Steve, Agbo Nelson Wheatson, Telfer C Trevor, Ross, Glenn Lindsay. Heavy Metal Concentrations and Sediment Quality of a Cage Farm on Lake Volta, Ghana. Aquaculture Research. 5 (2020): 2041-2051.\n8. Manyedi, Sechaba, William W. Anku, Ephraim M. Kiarii, and Penny P. Govender. Thermoelectric, Electronic, and Optical Response of Nanostructured Al‐doped ZnO@ 2D‐TiC Composite. ChemistrySelect 5 (2020): 13144-13154.\n9. Renu Kumari, Adeniyi Olugbenga Osikoya Adeniyi Olugbenga Osikoya, Francis Opoku, William Wilson Anku, Sudheesh Kumar Shukla, and Penny Poomani Govender. Composite 2D Nanointerfaces for Electrochemical Biosensing: An Experimental and Theoretical Study. ACS Applied Biomaterials. 12 (2020): 8676-8687.\n10. Onoyivwe Monday Ama, William Wilson Anku, Suprakas Sinha Ray. Photoelectrochemical degradation of methylene blue dye under visible light irradiation using EG/Ag-ZrO2 nanocomposite electrodes. International Journal of Electrochemical Science. 14 (2019) 9982-10001. \n11. Onoyivwe Monday Ama, Khotso Khoele, William Wilson Anku, Suprakas Sinha Ray. Photoelectrochemical Degradation of 4-Nitrophenol using CuOZnO/exfoliated graphite Nanocomposite Electrode. International Journal of Electrochemical Science. 14 (2019) 2893 – 2905.\n12. Ndaba, Nokuthula, Marthe Carine Fotsing, William Wilson Anku, and Penny Poomani Govender. In vitro and in silico studies of the antifungal properties of the bulb and leaves extracts of Drimia delagoensis Baker (Jessop). Advances in Traditional Medicine, (2019): 1-7.\n13. Samuel Osei-Bonsu Oppong, Francis Opoku, William Wilson Anku, Ephraim\nMuriithi Kiarii, Penny Poomani Govender. Experimental and Computational Design of Highly Active Ce–ZrO2–GO Photocatalyst for Eosin Yellow Dye Degradation: The Role of Interface and Ce3+ Ion. Catalysis Letters. (2019) 1-18.\n14. Renu Kumari, Adeniyi Olugbenga Osikoya, Francis Opoku, William Wilson Anku, Sudheesh Kumar Shukla, Penny Govender. Hierarchically assembled Two-dimensional Gold-Boron Nitride-Tungsten Disulphide nanohybrid interface system for electrobiocatalytic applications. Materials chemistry and physics, 226 (2019) 129-140.\n15. Madima Ntakadzeni, William Wilson Anku, Penny Poomani Govender, Leelakrishna Reddy. Mo3S4 nanorod: An effective photocatalyst for the degradation of organic dyes in aqueous solution. Recent innovations in chemical engineering, 12 (2019) 61-9.\n16. Madima Ntakadzeni, William Wilson Anku, Neeraj Kumar, Penny Poomani Govender, Leelakrishna Reddy. Pegylated MoS2 nanosheets: A dual functional photocatalyst for photodegradation of organic dyes and photoreduction of chromium from aqueous solution. Bulletin of Chemical Reaction Engineering & Catalysis, 14 (2019) 142-152.\n17. S. O.B. Oppong, W. W. Anku, F. Opoku, S. K. Shukla, E. S. Agorku and P. P. Govender. Photodegradation of Eosin Yellow Dye in Water under Simulated Solar Light Irradiation using La-Doped-ZnO Nanostructure Decorated on Graphene Oxide as an Advanced Photocatalyst. ChemistrySelect 3 (2018) 1180-1188.\n18. W. W. Anku, S. K. Shukla and P. P. Govender. Graft gum ghatti caped Cu2O nanocomposite for photocatalytic degradation of naphthol blue black dye. Journal of Inorganic and Organometallic polymers and Materials (2018) 1540-1551.\n19. C.N. Peter, W. W. Anku, R. Sharma, G. M. Joshi, S. K. Shukla, P. P. Govender. N-doped ZnO/graphene oxide: a photo-stable photocatalyst for improved mineralization and photodegradation of organics dye under visible light. IONICS (2018) 327-339.\n20. C.N. Peter, W. W. Anku, S. K. Shukla, P. P. Govender. Theoretical studies of the Interfacial charge transfer and the effect of vdW correction on the interaction energy of non-metal doped ZnO and graphene oxide interface. Theoretical Chemistry Accounts 137 (2018) 75-84.\n21. Renu Kumari, Adeniyi Olugbenga Osikoya, William Wilson Anku, Sudheesh Kumar Shukla, Penny Poomani Govender. Hierarchically assembled two-dimensional hybrid nanointerfaces: A platform for bioelectronic applications. Electroanalysis. Electroanalysis 30 (2018) 2339-2348.\n22. W. W. Anku, S. O. B. Oppong, S. K. Shukla, E. S. Agorku, and P. P. Govender. Cobalt doped ZrO2 decorated multiwalled carbon nanotube: A promising nanocatalyst for photodegradation of indigo carmine and eosin Y dyes. Progress in Natural Science: Materials International 26 (2017) 354-361.\n23. S. O. Oppong, W. W. Anku, S. K. Shukla and P. P. Govender. Synthesis and characterisation of neodymium doped-zinc oxide–graphene oxide nanocomposite as a highly efficient photocatalyst for enhanced degradation of indigo carmine in water under simulated solar light. Research on Chemical Intermediates 43 (2017) 481-501.\n24. W W Anku, S. O. B. Oppong, S K Shukla and P P Govender.Comparative photocatalytic degradation of monoazo and diazo dyes under simulated visible light using Fe3+/C/S doped-TiO2 nanoparticles. Acta Chimica Slovenica 63 (2016) 380-391.\n25. W. W. Anku, S. O. B. Oppong, S. K. Shukla, E. S. Agorku, and P. P. Govender. Chitosan–sodium alginate encapsulated Co-doped ZrO2–MWCNTs nanocomposites for photocatalytic decolorization of organic dyes. Research on Chemical Intermediates 42 (2016) 7231–7245.\n26. W. W. Anku, S. O. B. Oppong, S. K. Shukla, E. S. Agorku, and P. P. Govender. Palladium-doped–ZrO2–multiwalled carbon nanotubes nanocomposite: an advanced photocatalyst for water treatment. Applied Physics A 122 (2016) 579-587.\n27. W W Anku, S. O. B Oppong, S K Shukla and P P Govender. Influence of ZnO concentration on the optical and photocatalytic properties of Ni-doped ZnS/ZnO nanocomposite. Bulletin of Materials Science 39 (2016) 1745-1752.\n28. S. O. B. Oppong, W. W. Anku, S. K. Shukla, E. S. Agorku and P. P. Govender. Photocatalytic degradation of indigo carmine using Nd-doped TiO2-decorated graphene oxide nanocomposites. Journal of Sol-Gel Science and Technology 80 (2016) 38–49.\n29. M. Mzoughi, W. W. Anku, S. O. Oppong, S. K. Shukla, E. S. Agorku and P. P. Govender. Neodymium Doped ZrO2-graphene Oxide Nanocomposites: A Promising Photocatalyst for Photodegradation of Eosin Y Dye. Advanced Materials Letters 7 (2016) 946-950.\n30. S. O.B. Oppong, W. W. Anku, K. S. Shukla and P. P. Govender. Lanthanum doped-TiO2 decorated on graphene oxide nanocomposite: A photocatalyst for enhanced degradation of Acid Blue 40 under simulated solar light. Advance Materials Letters 8 (2016) 432-438.\n\nConference Presentations\n1.\tSession Co-chairs: William Wilson Anku and Saada Mohammed. Session Title: Innovative sample preparation and detection techniques for legacy and emerging pollutants in different environmental matrices. Virtual SETAC Africa 10th Biennial Conference held from 20-22 September 2021.\n\n2.\tW.W. Anku, S.O.B. Oppong, S. K. Shukla, E.S Agorku and P.P. Govender. Hetero-elements doped TiO2 for comparative photocatalytic degradation of monoazo and diazo dyes. SPEA9- 9th European Meeting on Solar Chemistry and Photocatalysis: Environmental Applications. Held in Strasbourg, France from 13th to 17th June 2016. \n\n3.\tW.W. Anku, S.O.B. Oppong, S. K. Shukla, E.S Agorku and P.P. Govender. Cobalt-doped ZrO2 decorated multiwalled carbon nanotube: A promising nanocatalyst for photodegradation of indigo carmine dye. 4th YWP-ZA Biennial Conference and 1st Africawide YWP Conference. Held at the CSIR-Pretoria, South Africa from 16th to 18th November 2015. (Won second best presenter award).\n\n4.\tW.W. Anku, S.O.B. Oppong, S. K. Shukla, E.S Agorku and P.P. Govender. Palladium doped-ZrO2-multiwalled carbon nanotubes nanocomposite as an advanced photocatalyst for water treatment. 5th UJ Cross Faculty Symposium. Held at UJ-Bunting Road Campus on 13th October 2015. (Won best presenter award).\n\n5.\tW.W. Anku, S.O.B. Oppong, S. K. Shukla, E.S Agorku and P.P. Govender. Cobalt-doped ZrO2 decorated multiwalled carbon nanotube: A promising nanocatalyst for photodegradation of indigo carmine dye. UJ Harvest festival. Held on 17 September 2015 in Perskor Building, DFC.\n\n6.\tW.W. Anku, S.O.B. Oppong, S. K. Shukla, E.S Agorku and P.P. Govender. Palladium doped-ZrO2-multiwalled carbon nanotubes nanocomposite as an advanced photocatalyst for water treatment. 3rd conference on Emerging Frontiers for Sustainable Water. Held at the Protea Hotel Wanderers, in Johannesburg, South Africa from 3-5 August 2015.\n\nReferences\n1. Prof Penny Govender\nDirector: Research Capacity Development (RCD)\nPostgraduate School: Research & Innovation, 101, Akanya Building\nAPK campus, University of Johannesburg, South Africa\nTel: 27845002689. Email: pennyg@uj.ac.za\n\n2. Dr. Anthony Yaw Karikari\nDeputy Director: \nCSIR-Water Research Institute, P.O. Box M38, Achimota-Accra, Ghana\nTel: 233208184215, E-mail: aykarikari@hotmail.com\n\n3. Dr Monday Onoyivwe Ama\nResearch Scientist: CSIR-National Centre for Nanostructured Materials,\nMeiring Naude Road Brummeria, Block 19B, Pretoria 0001, South Africa \nTel.: +27733300486, Email: onoyivwe4real@gmail.com',institutionString:"CSIR-Water Research Institute",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"2",totalChapterViews:"0",totalEditedBooks:"0",institution:null}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"14",title:"Materials Science",slug:"materials-science"}],chapters:[{id:"82929",title:"Prediction of Solubility and Miscibility Parameters of Bismuth-Arsenic Complex and Amorphous Mineral Compounds Using Molecular Dynamics Simulation",slug:"prediction-of-solubility-and-miscibility-parameters-of-bismuth-arsenic-complex-and-amorphous-mineral",totalDownloads:1,totalCrossrefCites:null,authors:[null]}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"440212",firstName:"Elena",lastName:"Vracaric",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/440212/images/20007_n.jpg",email:"elena@intechopen.com",biography:"As an Author Service Manager, my responsibilities include monitoring and facilitating all publishing activities for authors and editors. 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Kawsar Alam"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8417",title:"Recent Advances in Boron-Containing Materials",subtitle:null,isOpenForSubmission:!1,hash:"3737be3f785ef9d8b318571ab474f407",slug:"recent-advances-in-boron-containing-materials",bookSignature:"Metin Aydin",coverURL:"https://cdn.intechopen.com/books/images_new/8417.jpg",editedByType:"Edited by",editors:[{id:"27070",title:"Prof.",name:"Metin",surname:"Aydin",slug:"metin-aydin",fullName:"Metin Aydin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8812",title:"Contemporary Topics about Phosphorus in Biology and Materials",subtitle:null,isOpenForSubmission:!1,hash:"86c427901f631db034a54b22dd765d6a",slug:"contemporary-topics-about-phosphorus-in-biology-and-materials",bookSignature:"David G. Churchill, Maja Dutour Sikirić, Božana Čolović and Helga Füredi Milhofer",coverURL:"https://cdn.intechopen.com/books/images_new/8812.jpg",editedByType:"Edited by",editors:[{id:"219335",title:"Dr.",name:"David",surname:"Churchill",slug:"david-churchill",fullName:"David Churchill"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6851",title:"New Uses of Micro and Nanomaterials",subtitle:null,isOpenForSubmission:!1,hash:"49e0ab8961c52c159da40dd3ec039be0",slug:"new-uses-of-micro-and-nanomaterials",bookSignature:"Marcelo Rubén Pagnola, Jairo Useche Vivero and Andres Guillermo Marrugo",coverURL:"https://cdn.intechopen.com/books/images_new/6851.jpg",editedByType:"Edited by",editors:[{id:"112233",title:"Dr.Ing.",name:"Marcelo Rubén",surname:"Pagnola",slug:"marcelo-ruben-pagnola",fullName:"Marcelo Rubén Pagnola"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"81708",title:"High Throughput Methods to Transfer DNA in Cells and Perspectives",doi:"10.5772/intechopen.104542",slug:"high-throughput-methods-to-transfer-dna-in-cells-and-perspectives",body:'The most used approach to decipher proteins’ function or their interactome is to study the effects induced by the delivery of exogenous materials in living cells (deoxyribonucleic acid: DNA, ribonucleic acid: RNA, oligonucleotides, proteins, and ribonucleoproteins). Coding sequence overexpression, then gene silencing, and genome editing approaches offer a panel of induced biological modifications within cells that allowed us to increase our knowledge of most cellular processes. However, in a post-genome era, thousands of genes must be studied and exogenous material transfer into cells, including DNA, became a limiting factor. Indeed, available technologies predominantly allowed analysis at a gene-by-gene scale, and new approaches were developed to reach higher throughput. Libraries of material such as small interfering RNA (siRNA) [1, 2] and Open Reading Frame (ORF) expressing plasmids collection were developed [3, 4] to cover all proteome. To take advantage of these, concomitant High-Throughput (HT) technologies are pointed out for their transfer in cells. Plasmid DNA (pDNA) transfer in cells (by transfection or transduction) plays a central role when studying the precise biological role of proteins. For pDNAs, several efficient transfection methods were pushed to higher throughput. All these induced changes performed in cells allow not only our understanding on the biological processes of cells’ life but also have therapeutic applications [5, 6]. The huge interest in gene and cellular therapy approaches is indeed a motor in the development of highly efficient gene delivery strategies.
In this chapter, we will first give a brief overview of DNA transfer methods in cells, then a more detailed part will focus on those that reached higher throughputs and we will conclude with future expected enhancements.
To promote a biological effect in the cell, exogenous DNA must face several levels of pitfalls starting from the outside of the cell. First, it must cross the plasma membrane composed of a hydrophobic lipid bilayer which naturally prevents hydrophilic material such as DNA from entering the cells. In addition, DNA and the plasma membrane carry a general negative charge that impedes DNA transfer into cells by electromagnetic repulsion. Furthermore, once entered, the DNA has to face degradation mechanisms that occur in the cells. Finally, if part of the exogenous DNA succeeds in passing all these steps, the expected biological effect would be measurable. To circumvent all these, a range of approaches to transfer DNA has been developed. DNA delivery to cells can be divided into three main categories: physical, chemical, and biological methods. Among all approaches, viral ones are the most efficient but present some limitations such as the transgene size and biosafety issues. Physical and chemical methods were developed to circumvent these limitations and are not limited in the size and number of genes to be transferred. Some of these are still largely used whereas some were more a proof of concept. In this section, we briefly describe the physical, chemical, and biological methods.
As mentioned above, due to the plasma membrane and DNA respective properties, the transfer of DNA into cells is impaired. All the physical methods aim to directly circumvent the hydrophobic and electrochemical repulsion parameters by disrupting the integrity of the membrane and promoting a transient permeability. Physical methods then do not have a limit in cargo size and do not depend on biological mechanisms as a direct material delivery is performed [7].
An evident method is the direct microinjection of DNA in cells, which was performed in early-stage embryo [8], and then human cell lines [9]. It implies micromanipulation of a single cell under a microscope to bypass the membrane barrier using a thin glass needle to inject DNA directly into its cytosol or compartments [8, 10]. This approach is reproducible but tedious due to the need to inject each cell individually.
Electroporation method has emerged when it was shown that an electric field could promote a loss of membrane permeability by transient pore formation [11] thus allowing DNA delivery in cells [12]. DNA, target cells, and electroporation buffer laying between two electrodes are submitted to an electric pulse [13]. This pulse is divided into a high-voltage stage to create temporary pores, and a low-voltage one to allow electrophoresis of DNA through these pores [14]. Extensive optimizations (pulse voltage and duration, buffer composition) were done to balance transfection efficiency with cell viability as the requested high voltage promotes cell death [15, 16]. “Electroporators” devices are nowadays available with many predefined settings to achieve efficient transfection in almost all cell types, even hard-to-transfect ones [17].
Biolistic or micro-projectiles bombarded to the cells represent another delivery mode. The projectiles made of gold-covered by nucleic acids, penetrate into cells by high-speed bombardment [18]. First developed for plants, this approach is also efficient in mammalian cells/tissues [19]. However, the method suffers the cost of particles. Nanoparticles that can bind nucleic acids, and whose small size allows them to pass cell membranes with high efficiency, represent a cheaper alternative [20].
Femtosecond laser optoporation consists in focusing ultrashort laser pulses on a cell membrane to induce a transient perforation. This membrane perturbation allows the pDNA transfer [21]. Many cell types can be transfected using a variety of laser sources [22, 23]. Despite efficient, due to the needed laser focusing on a single cell level, its throughput is limited.
Acoustoporation or sonoporation uses ultrasounds to induce a transient plasma membrane disruption promoted by bubbles cavitation phenomenon and thus allowing gene transfer [24, 25]. The method was enhanced by the use of high-frequency waves creating reversible nanopores and furthermore promoting “molecular bombardment” on the bilayer membranes that enhances DNA delivery while limiting cell mortality [26].
Passing constriction or nano-constriction is an approach based on the mechanical deformation of cells as they pass through micro constrictions channels [27]. This controlled compression induces transient pores formation into the cell membrane and allows DNA entry from the surrounding buffer [27]. This method is expected to be universal and showed efficiency in easy and hard-to-transfect cell lines like primary and stem cells [28].
The last method, magnetofection, has been classified as a physical method. It is based on magnetic nanoparticles (MNPs) coated with transfection reagents that bind nucleic acids and promote cell entry [29]. Indeed, MNP only induces the concentration of the MNP on the cells mat when a magnetic force is applied but is per se not able to transfer DNA into cells. However, it enhances DNA delivery up to several hundred and allows to lower DNA consummation, and is furthermore efficient in hard-to-transfect cells [30, 31].
Interest to develop non-viral and reproducible gene delivery methods has led to the use of chemical reagents. Chemical transfection methods represent an alternative way to bypass the membrane barrier and furthermore try to protect DNA from degradation within cells [32]. These reagents promote DNA compaction, negative charge neutralization, and cell interaction for later entry into cells. These reagents are briefly summarized here after.
Calcium phosphate co-precipitation is the cheapest method and was first described in 1973 [33]. It relies on the formation of a precipitate when the negatively charged DNA binds to calcium ions (Ca2+) [34]. This precipitate interacts with the plasma membrane and enters the cell by endocytosis [35]. This widely used method reaches up to 90% efficiency for easy-to-transfect cells but is impaired by the need for fresh preparation, avoiding any storage of ready-to-transfect plates [36]. The formation of an efficient precipitate depends on several parameters and this method can be toxic for certain cells such as primary ones [37]. Calcium was also shown to enhance gene delivery by other methods [38] and was then tested alone as a transfection reagent (calfection) [39]. The mechanism does not rely on the formation of a precipitate and do not need fresh preparation. Furthermore, the Ca/DNA mixture can be stored for a long period without any loss in efficacy. Intended for batch transfection of the high number of cells, it worked in a 12-wells plate format for adherent or non-adherent cell lines. The easy use, storage ability, and low cost make this method interesting whereas it was not tested so far in higher throughput.
The diethylaminoethyl-dextran (DEAE-dextran) is another reagent that showed efficiency [40]. This polycationic derivate of dextran compacts DNA to form a positively charged complex that later interacts with the plasma membrane to enter cells by endocytosis [41]. The method is simple, low cost, and efficient for many cell types however, new enhanced approaches surpassed it.
Lipofection method is based on the use of lipids and cationic lipids [42, 43]. When mixed with DNA solution, these lipids form liposomes, a kind of vesicular structure with the same composition as cellular membranes and entraps DNA in solution [44]. The formed complexes (lipoplexes) allow DNA delivery through binding to the cell membrane (due to electrostatic forces), cell entry, mainly by endocytosis [45], and release of the DNA for expression. Lipids-based transfection reagents are efficient and mostly insensitive to serum so that medium has not to be removed before transfection. Furthermore, lipofection can be used efficiently in forward or reverse mode transfection in numerous cell lines [46]. Cationic lipids are more and more efficient in DNA delivery, and furthermore efficient on suspension or adherent cells, and for increasing number of cell types, and even hard-to-transfect ones [47].
Cationic polymers are non-lipidic as deprived of a hydrophobic moiety and are then soluble in water. They use a similar mechanism: being positively charged, they interact and compact DNA under the form of polyplexes [48]. They enter the cell by endocytosis, and traffic through endosomes and cytoplasm to finally deliver DNA to the nucleus [49]. This class of reagent has the advantage to limits DNA degradation in lysosomal compartments, increasing delivery efficiency [50].
Biological approaches to transfer DNA are inspired by natural mechanisms. The most potent of these approaches is gene transfer by viruses. Other methods represent fields in expansion: cell-penetrating peptides or the use of exosomes or vesicular transfer. These approaches do not rely on natural products but on diverted forms to allow the transfer of a gene of interest.
Viral approaches are the highest efficient among all, even in hard-to-transfect cells [51]. To be permissive, the cells must express the receptor interacting with the virus envelope proteins. To enter in almost all cell types, a ubiquitous and widely expressed receptor is preferred. The Vesicular Stomatitis Virus G (VSV-G) protein promotes entry in almost all cell types as the Low-Density Lipoprotein Receptor family is its ubiquitously expressed receptor [52]. Its interaction with the VSV-G protein promotes membrane fusion and allows virus content to be delivered to the cells [53]. The use of a viral vector is however limited in throughput as viral particles have to be produced for each different DNA to transfer. This production involves the cloning of the gene of interest in a viral vector backbone that is later transfected into a packaging cell line to be integrated into pseudo-viral particles. Pseudo-virus are then recovered from the cell’s supernatant, concentrated, and titrated before their use for transduction of the target cells. Despite lower throughputs, viral delivery remains the most powerful way to transfer DNA in cells, even in primary cells (90% efficiency).
The fusiogenic envelope G glycoprotein of the VSV-G was also used as a reagent for gene transfer when mixed with plasmid DNA [54]. The resultant product termed “Gesicles” showed 55% transfection efficiency in HeLa cells, and 22% for hard-to-transfect human myoblast cells [55]. Whereas promising, this method did not reach HT yet.
Another interesting biological derivative used for DNA transfer is represented by proteins having natural properties to enter the cells by surface receptors dependent [56] or independent mechanisms [57]. Some natural peptides derived from these proteins, the cell-penetrating peptide (CPP) are able to enter the cell through the membrane [58, 59]. These peptides have short lengths and a global positive charge. Involved mechanisms are still unclear and depend on the CPP (direct penetration, endocytosis, or translocation via intermediate structure in the membrane lipid bilayer). Peptide from the Trans-Activator of Transcription (TAT) protein of the Human Immunodeficiency Virus (HIV) was efficiently used as a DNA carrier in HeLa cells [60]. Some others have been modified and their properties mixed with each other to promote efficient delivery of exogenous nucleic acid into cells [61]. CPP can be engineered by multiplexing peptides with distinct properties or by modifying their composition. One of the engineered CPP is the pepFect14 [62] which showed efficiency for DNA delivery in several cell types such as CHO, HEK293, U2OS, or U87 cells [63].
One last example of naturally occurring biological derivatives is the use of exosomes. First described in 1977, these nano-sized vesicles derived from plasma membrane elements, are involved in mediating messages to proximal and distant cells [64, 65]. This natural process is found in normal or pathological cells [66] and can be turned around to deliver DNA of interest [67].
To enhance the throughput of the experiments performed on cells transfected by exogenous DNA, it is interesting to do it in a HT way. However, a distinction must be done between experiments performed at HT using transfected cells, and HT transfection of cells. Indeed, depending on assay requirements, transfection of a single condition may be performed using a large volume of suspended cells that are then distributed among several individual wells for subsequent treatments and assays [68]. Alternatively, it is interesting to transfect many different plasmids, each well of transfected cells expressing different transgenes [69, 70]. This difference is generally concomitant with the way the transfection is performed: batch protocol or not.
Batch protocol allows to transfect a large number of cells that are then dispatched in separate wells for further experiments. In this case, all transfected cells in the batch share the same conditions of transfection. This protocol is generally used to limit variability in HT assays for monitoring the effect on a biological parameter under a unique transfection condition. Typically, it can be performed on adherent cells in a forward-protocol modus: cells are plated and transfected 24 h later, according to the transfection reagent’s manufacturer instructions. The day or several hours after transfection, adherent cells are suspended and dispatched in multi-well plates (96, 384, or even 1536) for further HT treatments and analysis [71]. Depending on the cells used, the batch transfection is also compatible with the suspended cells that are then directly dispatched on separate wells after transfection.
The batch protocol is not per se a HT transfer of different biological materials in cells, but rather a way to perform HT assays and treatments in separate wells. On the opposite, HT protocols can achieve a true HT transfection in which each well receive a different DNA or transfection conditions.
To be able to determine the behavior of cells or biological effects induced by the transfer of many different pDNAs in the cells, a real HT transfection becomes interesting. Several methods allow the management of numerous pDNA or different conditions when transfecting the cells, but to reach HT, good efficiencies are almost necessary. HT transfection can be achieved using several methods that are presented below.
As described before, electroporation is performed with buffer diluted cells and DNA, subjected to an electrical pulse that promotes membrane destabilization. Many devices, protocols, and dedicated buffers have been implemented to reach universal use. However, it seemed incompatible for HT as each separate transfection must be performed one by one in micro cuvettes. This problem has been solved by the development of new devices able to deliver an electrical pulse simultaneously in several wells on dedicated plates. Harvard Apparatus/BTX developed an up to 96 wells approach using plates with embedded aluminum electrodes. Used with the plate handler Model HT-200, it allows transfection in 8 wells simultaneously and was shown efficient in neurons [72].
Another approach based on an array of 96 suspended electrode pairs fitting on top of standard 96-well plates represents a less expensive approach [73]. Each pair of electrodes can be loaded and held 10–20 μL of transfection mixture by the surface tension. After pulse delivery, the direct addition of the cell culture medium into the array allows the electroporated cells to drop and seed into the underlying microplate. The array is reusable, and uses standard microplates and inexpensive standard buffers, reducing the cost of this approach. In addition, common liquid handling robots can achieve a 96-well transfection time of approximately 1 min. This technology could be adapted to the 384-well plate format using a more sophisticated electrode array design and concomitant robotics.
Whereas successful in almost all cell types, the electroporation method has some limitations. First, it is the most DNA-consuming one of all the HT transfection approaches described so far. Secondly, as a cell suspension is required during the electrical pulse delivery, it avoids its use on adherent differentiated cells mat. Nevertheless, its advantage in terms of success in almost all cell types, and its versatility concerning the material to transfer (not only efficient for DNA) promises electroporation to further future enhancements and use.
In 2001, Amaxa™, (now owned by Lonza™) launched an electroporation-derived method termed nucleofection as DNA is transferred directly into the nuclei of the cells. It shortens the time of experimentations, by suppressing the necessary nuclear import step of DNA and ensure proper expression of transgene [74]. It relies on an electroporation-based device (Nucleofector) and the use of dedicated buffer solutions to ensure nuclear transfer. The exact mechanism allowing nucleus targeting and buffer composition is kept proprietary. However, since the first published results on natural killer cells transfection [75], it has been widely used in many hard-to-transfect cells with efficiency ranging from 25 to 70% [76]. First, nucleofector devices used nucleocuvettes and were then limited in throughput. New apparatus and dedicated consumables were developed to reach higher throughput: the 96-well Shuttle® device (amaxa AG), in which cells are plated on 96 wells “nucleocuvette plates” and pulsed using Nucleofector™ programs. These plates, made of conductive polymers, allow the current delivery in each well individually. It takes less than 10 min in an automated way to process the entire plate [77]. Many optimized conditions have already been defined using nucleocuvettes depending on cell types (programs for the electrical pulses, cells number, and optimal buffer conditions) and as an advantage, these settings are transposable to nucleocuvette plates [17]. Numerous successful examples have been published ranging from 35 to 70% efficiency: primary chondrocytes [78]; dendritic cells [79], and even H9 hESC [80].
To push further the throughput a 384-well Nucleofector™ requiring 384-well Nucleocuvette™ plates was launched. The complete electrical pulse delivery process takes just one minute, and several wells are processed at the same time [81]. However, the overall process is the same as for the previous model, mixing of cells with buffer and DNA in the wells, delivery of the electric pulse, the addition of fresh medium in the 384-well Nucleocuvette™ for cell recovery, and then their dispense in a cell culture plate for later experimentations.
Whereas versatile and being efficient as electroporation in many cell types, nucleofection is still restricted to suspended cells, impairing its use on morphologically differentiated and adherent cells. Furthermore, the need to transfer transfected cells to a standard plate for further experimentation is a limiting step of the method. The cost of such approaches broadens their wide use in the scientific community due to the price of transfections kits, containing ready-to-use buffers, and nucleocuvette plates.
As explained above, electroporation or nucleofection are restricted to cell suspensions. To circumvent this limitation, new kind of electrodes able to deliver the electric pulse on a cell mat was developed.
One of the simplest developed approaches is an electrode device that takes place on top of standard culture cell dishes. The PetriPulser™ (from BTX) consists of 13 gold plated electrodes embedded in an isolating holder placed above the Petri dish containing the cell mat to electroporate [82]. This model fits 35 mm Petri dishes but a scaled-up model, the “Petri dish electrode” made of stainless steel electrodes, fit 100 mm diameter dishes [83]. The 2 mm distance between electrodes is the same as in most cuvettes. A model for transwell cultured cells electroporation: the BTX™ Adherent Cell Electrodes [84] presents a 5 mm distance inter electrodes and may engender adverse effects on cell viability. All these devices are reusable, lowering the cost of this approach that has however not been used so far in published works.
A sophisticated version was launched by Cellectricon™: the Cellaxess®HT. It uses dedicated 384-wells microplates and a capillary embedded microelectrodes array. Using a platform device, adherent cells seeded in 384-wells plates are washed, electroporated using transfection mixture (loaded from side donor plates), and allowed to recover with fresh medium addition. 96 wells are simultaneously electroporated by the device and throughput of 50,000 wells per day is announced by the manufacturer [85]. However, it was not really used in the academic laboratories as no work has been published except the proof of concept of the manufacturer. They simplified the method by launching the Cellaxes Elektra-Adherent Cell Electroporation System. It is also an electrode-based electroporation system optimized for the
Array approaches are based on spotting an array of transfection reagents and material to transfer on a planar slide where cells are later plated. Using such approaches with electroporation method was unimaginable. However, several teams pushed down this restriction by developing custom-made devices to electroporate adherent cells in a microarray manner. Two technologies are suitable for adherents cells: the delivery of the electrical pulse between the bottom and top of chip micro-wells; or between interlaced microelectrodes laying on the bottom of the dishes under the seeded cells [87, 88].
In the HT
Another method was able to electroporate adherent cells, based on a glass gold electrode coated with PEI for pDNA loading [91]. Cells are plated on this electrode and the electrical pulse can be delivered using an additional top cover electrode up to 3 days post-seeding. Transfection efficiency reached 90% in HEK but was also efficient in primary fibroblasts. Although electroporation was performed in 13 mm square areas, this method allowed HT transfection using up to 169 plasmids micro-arrayed on the electrode. This method seems affordable, as it only requires a gold vaporized electrode.
Whereas it remains a field of specialists, microfluidic applications increased in the last decade due to their low-cost advantage, as it can be in-house designed using affordable technologies, and it deals with low quantities of reagents. Microfluidic can manipulate different solutions and mix them, and lead to cell culture and transfection chips design [92]. However, in-house designs might be difficult to reproduce, even more, if highly specialized skills are required. Furthermore, most biological experiments require a subsequent amount of transfected cells, harder to achieve using microfluidic. Despite these limitations, success in microfluidic transfection applications has been published for a wide variety of cells, and even at the single-cell level [93, 94]. First devices lacked the necessary throughput to test numerous transfections conditions in parallel, but recent advances pushed it further. In the field of transfection, two main approaches have been used with microfluidics: electroporation and nano-constriction.
Electroporation in standard 2 mm cuvettes requires high voltage that promotes cell death by a joule heating effect, a local pH change due to water electrolysis, that in turn induces the formation of bubbles promoting cells aggregation and impairing the DNA delivery efficiency [95]. Due to its efficiency, electroporation was used in microfluidic derivatives trying to circumvent some of its limitations. Embedding electrodes in a microfluidic channel can limit adverse effects on cell viability [92]. The diameter of the channel allows the electrodes to be closer to each other’s and the use of voltages as low as 1 volt [96], reduces the heating joule effect, electrolysis, and bubbles. pH modifications are still present but enhanced buffer composition improved it [97]. These microfluidics devices mostly use flowing cells transfected in a semi-continuous way [98], avoiding testing many different conditions in parallel and lowering throughput. Some devices allow transfection of adherent cells in micro-chambers using a porous substrate on which cells are seeded. The electric field is then applied through the cells (under/upper compartment). This has been successfully performed on stem cells differentiated in neurons [99]. Despite the latest improvements, microfluidic-based approaches still lack HT. However, due to the booming application of microfluidic, reaching higher throughput would be achievable and a promising way to perform transfection.
Most of the chemical transfection reagent allows two kinds of protocols: the forward and the reverse protocol. In forward protocols, DNA and transfection reagent are mixed to form transfection complexes and then distributed on previously seeded cells. Such an approach is harder to manage in a HT way as each different mixture condition implies a different container (tube or wells of multiplate wells) and necessary tedious pipetting steps. However, this kind of protocol can be manually achievable with standard molecular biology material such as multichannel micro-pipettors. An experimented user can transfect one to four 96 well plate manually in 2 h with up to 3 different pDNAs per condition [100, 101]. However, to our knowledge, the forward approach has not been automated so far to reach HT.
The forward mode has been surpassed by the reverse protocol mode. The DNA (eventually with the transfection reagent) is directly dispatched on the final wells (i.e., of a multi-well plate) or a glass slide, and cells are added directly on these deposits. This mode of transfection has several advantages: first it shortens the overall experimental time, second, it can easily be automated allowing to reach HT and good reproducibility. Suitable for such an application, liquid handling devices enable the dispense of low liquid volumes for the multiplexing of different solutions whose concentration and ratio are tightly controlled in each well. Such protocols have been developed for most of the biological material to transfer which includes DNA and follow the technological developments available to do it. An overview of these methods used for DNA transfection in a HT manner is detailed below.
As mentioned before, lipidic transfection reagents are eligible to reverse protocol, making them suitable for potential HT approaches. This reverse mode was shown efficient on CHO cells grown in suspension in a 96 wells-plate format using PerFect Lipids (pFx-6 form lnvitrogen) as reagent [100] and even adherent cells using Lipofectamine (Invitrogen). Higher Throughput was reached using Turbofectin8 as reagent (Origene) and plasmids coding 704 different transcription factors dispensed in 384-wells plates [102].
The SMAR-chip described before in the HiCEP method [89], was also applied to HT reverse transfection but using Lipofectamine 2000 as a transfection way instead of electroporation. It allowed the efficient transfection of HEK293 (up to 65% transfected cells) in the 169 wells of the matrix. The authors aimed at producing viral particles using co-transfection of the necessary plasmids with 169 genes of interest. Proper viral packaging and sufficient viral production were shown by successful transduction of side cultured 3T3L1 cells using the supernant of the HEK producing cells.
Tavernier’s group reached a much higher throughput in 2002 using reverse transfection for HT transfection of HEK293 cells in its MAmmalian Protein–Protein Interaction Trap (MAPPIT) Arrays approaches to study protein–protein interactions [103]. Effecten reagent was used in a reverse mode protocol to transfect prey expressing plasmids in up to 384-wells plate format using classical liquid handling facilities and a mammalian ORF collection plasmids.
With the emergence of such collection, examples of microplate-based arrays of the huge collection of plasmids have grown. One of the highest throughput was reached using 6049 different human cDNA expression plasmids to study their effect on the promoter activation of the zinc-finger protein RP58, using a luciferase reporter gene [104]. 50 ng plasmid/wells were loaded on sets of 384-well plates and a HT reverse transfection of HEK293 was successfully performed using Lipofectamine 2000.
A HT transfection protocol was reached in 384-wells plates format using non-liposomal polymers (Mirus TransitX2) as transfection reagent [101]. A reverse protocol led to about 90% transfection efficiency (even in cotransfection assay). The originality of this work is the use of a tips-free acoustic delivery of reagent and DNA (Echo nanodispencer from Labcyte™). This device sends multiple droplets of 2.5 nL from a 384-wells source plate to a destination one up to 1536-wells plates. Starting from unique diluted plasmids solutions, the overall process takes less than 20 min for one plate, and transfection ready plates can be stored dry or frozen without loss of efficiency. Cells are seeding on dry or freshly dispensed plates in a reverse mode transfection. The optimized protocol would allow 20,000 human genes transfection in about 18 h on a dedicated automated platform. Nano-quantities of DNA and reagents should render this approach low cost if the nanoaccoustic dispenser was not such expensive. Nevertheless, this protocol renders transfection affordable for newbies as the tedious work of DNAs and reagents combining in each well is controlled by spreadsheet driven software [105].
In 2001, DNA transfection throughput was pushed further by the use of a microarayer for the generation of transfection ready arrays of DNA [106]. In this study, 140 different plasmids DNA/gelatin mixture were deposited on glass microscope slides as 1 nL spots (of about 150 μm diameter). Effecten, a lipid transfection reagent was used to transfect cells seeded on the overall slide. Each spot led to the transfection of 30–80 HEK cells, in a DNA dose-dependent manner from 10 to 50 pg. Storage of the dried glass slides for more than 3 months did not affect transfection efficiency, allowing the matrix to be prepared in advance of use. Since this princeps study, other groups have successfully used this approach. Using the same reagent, one study transfected 16 different plasmids expressing proteins to study their cellular localization [107]. Another group used this approach for the HT screening of potential therapeutic membrane-displayed single-chain antibodies [108]. A true HT attempt was reached by the use of 1959 un-tagged ORF taken from the Mammalian Gene Collection (MGC) and expressed in HEK cells to identify genes implicated in apoptosis [109]. One similar array approach showed efficiency using Lipofectamine 2000 directly in the DNA mixture before arraying [110]. However, whereas simplified by combining the transfection reagent with DNA before dispensing, it requires about 10-fold more DNA to reach the same efficiency as the above protocols. A throughput of 2880 conditions on a complete 96-wells plate to study v-Src Mutant Protein Function was reached in HEK cells, using 30 spots of pDNAs mixtures per well of 96-wells plate [111]. Lipofectamine 2000 also showed efficiency in another microarray approach testing 600 cDNA spots on a single glass slide using reverse transfection [112]. Authors showed high efficiency in many cell types such as mouse preadipocytes (3T3L1), muscle myoblasts (C2C12), liver hepatoma (Hepa1c1c7), or macrophage (RAW-164.), human cervix epithelia adenocarcinoma (HeLa), or at bone osteosarcoma (UMR-108).
Tavernier’s group also pushed further its MAPPIT and MAmmalian Small molecule-Protein Interaction Trap (MASPIT) microplate-based array to microarrays using attractene (Qiagen), a non-liposomal lipid, as transfection reagent and a fluorescent reporter gene in place of the initial luciferase reporter [113]. Here, the ORFeome derived prey plasmid collection (15,000 cDNA) and a fluorescent reporter plasmid was mixed in 384-wells plates used as a matrix for further depositing by a microarrayer on polystyrene plates, to reach an industrial scale.
All these arrays’ methods are impressive in terms of throughput as many conditions, or different expressed genes, can be tested simultaneously in parallel cells. However, they require a consequent preparation time. DNA dilution, most of the time with gelatin, and optimally with the transfection reagent are generally performed in 96 or 384-wells plates. Once done, an arrayer robot is then plunging its tips for deposition of the DNA on several slides. The tips must be washed with detergent and then sonicated or heated to avoid cross contaminations before arraying the next DNA mixture. Finally, when the full array is printed, the slides have to be dried for 12 h to 2 days before later use and cell seeding. At the end of the experiment, a slide scanner became necessary to analyze transfected cells. The real throughput of such methods is then truly high once the arrays are ready to be incubated with the cells. Once the reagent used is efficient with the cell type requested, the throughput becomes only dependent on the liquid handling facility available in the lab. However, the method needs a certain financial investment for robotics, microarrayer platform and a scanner as the spots size and inter-distance need high resolution scanning to be analyzed.
As previously mentioned, microfluidic is now widely used due to the miniaturized scale it allows. Whereas it was applied to transfection using nanoconstriction or electroporation, it can also be used as a liquid and cell manipulation tool to perform transfection using chemical reagents. Schudel et al. first developed an inexpensive microfluidic-based miniaturized RNAi screening platform [114]. It relies on the use of a lipid-based transfection mixture and is low throughput as a maximum of 8 parallel transfections can be performed on this chip.
In another study, a two microchannel irrigating 8-chambers was designed on a glass slide [115]: 10 nL of a reverse transfection mixture containing gelatin, fibronectin, Lipofectamine 2000, and plasmid DNA were arrayed on a coated glass slide. This slide is mounted under the microscope, to face the microfluidic embedded chambers and showed successful transfection of Cancer LBT-N2b cells with almost no induced mortality, but the throughput was still clearly limited.
Based on the same kind of chambers design, the highest throughput was reached with a microfluidic chip of 1.6 × 5.8 cm containing 280 separate chambers. In about 10 min, the complete chip is loaded with about 600 cells per chamber of 500 μm diameter [116]. A set of valves allows the loading of different cell densities or even cell types. Once cells are loaded, the functional chip is obtained by alignment of the chambers to 280 DNA arrays (Lipid-DNA transfection mixtures) spotted on polylysine matrixes in an automated manner (about 2 h to complete). The assays showed a high transfection rate (99% efficiency) using an optimized condition but a cell line-dependent optimization is necessary. Whereas feasible, microfluidic managed reverse transfection still seems to have a long road to meet the scientific community mostly due to its required skills in the field, to be able to reproduce or use such devices.
As discussed before, some natural biologicals materials, viruses, proteins, peptides, or macromolecules have shown cell-penetrating properties and their ability to deliver different molecules to target cells either in their natural form, modified, and sometimes multiplexed by engineering. Here are some examples of such approaches that reached HT in the delivery of DNA into cells.
The main limiting factors to reach HT with viral delivery is the ability to produce these particles (i.e., biosafety cabinets class 2 or 3), in a HT manner (one independent viral production for each cDNA to transduce), and at a sufficient titer to promote efficient transduction of target cells. This production step has been shown feasible at HT in a pilot study with 1990 ORFs from the mammalian ORFeome collection [117]. In an automated platform, HEK cells were reverse co-transfected with these “gene of interest” plasmids and viral packaging ones to allow the production of the corresponding lentivirus in a 96-wells plate format (viruses transferring one ORF per wells). Supernatants (cDNA containing viral particles) were used to transduce target cells seeded in 96-wells plate format. In a similar manner, up to 16,000 cDNA were pushed to HT lentiviral production in 96-wells plates for later HT expression in target cells [118].
The previously developed SMAR-chip [89] was also used for viral particles production on the 169 matrixes embedded microwells, using reverse lipofection [119]. The method showed sufficient production to transduce 3T3L1 cell cultured in parallel to the producing cells array.
Despite these advances, such methodologies remain difficult to settle routinely due to the required material, specific skills, knowledge, and adequate biosafety facilities. To render it more accessible, some companies now propose ready-to-use kits in 96-wells microplate format to produce viruses in high titers from lab collection of cDNA [120]. Despite these limitations, this approach is highly promising as being universal for almost all cell types with high efficiency and furthermore efficient on suspension or adherent differentiated cells.
One example of protein derivatives used is collagen derivatives, which are produced by collagen treatment or digestion. Atelocollagen, is a polymer obtained by pepsin treatment of type I collagen that shows various effects in cell and animals. Atelocollagen condenses and delivers DNA, antisense oligodeoxynucleotides, or siRNAs into cells on its own [121]. Protocol based on this polymer reached a HT microplate array level in 2001, with a collection of pDNA showing a long-term gene expression in HEK cells [122]. The array can handle long storage without loss of efficiency. Another study reached HT transfection in PC-12 cells using Atelocollagen and 288 different plasmids dispensed in 96-wells microplate arrays [123]. The advantages of these last approaches remain in the fact that atelocollagen intrinsically regroups two properties in a single bio-product: DNA condensation and cell entry of the formed complexes into cells. Furthermore, it is derived from a biocompatible natural material and per se is rarely cytotoxic for cells.
Due to their potential, the use of CPP was pushed to HT transfection. The surface transfection and expression protocol (STEP method) is the only biological derivatives-based DNA transfer approach that reached such HT. It relies on the use of transferrin receptor, polylysine, adenoviral penton protein, and the HIV Tat protein to engineer some chimeric proteins. These combine functional motifs: binding of the DNA, binding to cell-surface receptors, the facilitated passage across membranes, the DNA targeting to the nucleus, and also adhesion and survival of the target cells on the arrayed spots [124]. The DNA/recombinants proteins mixtures are loaded in 384-wells source plates for standard arraying. Optimized conditions showed efficient GFP plasmid transfection efficiency (50–80%) and transgene expression in several cell types from easy to transfect HEK cells to more difficult ones such as SH-SY5Y neurons, N2A neurobalstoma cells, or PC-12 pheocromocytose cells. This method is promised for future enhancements accompanying the study of new CPPs. Indeed, many CPP have already been identified and validated leading to the creation of a dedicated database in 2012 referencing 843 CPP identified so far [125]. However, an exhaustive list is impossible to give as some are still identified nowadays and the developed database now contains 1700 unique CPPs 10 years later [126]. Some of them may represent better candidates for DNA transfer. This DNA transfection approach is also of great interest for gene therapy as it enables a kind of transduction, efficient like viral particles but without all the safety concerns for their production and use [127].
Last human genome sequencing assembly led to more than 24,000 genes to study [128]. Many approaches to transfer DNA in cells were then pushed to HT to interrogate each gene function. While still in progress with developments of new reagents and methods, HT DNA transfer approaches are already available. The main remaining challenge is to render them cheaper and affordable for non-specialists.
Among physical approaches, electroporation methods surpass the others being efficient in all cell types. The suspended electrodes array design represents several advantages: it is low cost, usable with standard electroporators and liquid handling devices, but is currently limited to 96-wells plate format [73]. Due to the technical design, it should be amenable to a 384-wells plate format. However, it is still restricted to suspended cells. Electroporation approaches for adherents’ cells have also been developed in 384-wells plate format, but suffer from their cost and their need for expensive consumables [86].
Microfluidics devices suffer from the required skills and technologies to be assembled and used. Microfluidics combined with electroporation appears as a solution to some limitations but chamber-based devices seem too far from the standard assays format to be widely used. Applicable to all cells, microfluidic devices based on semi-continuous electroporation of flowing cells currently lack the necessary throughput [98]. The same concern is pointed out for nanoconstriction-based transfection designs [28]. However, higher throughputs would be amenable as microfluidic manipulation of cells and solutions in an automated way is possible at a high rate. Such a device would advantageously require an automated loading of pDNA from a source plate to the chip, transfection of the expected amount of cells and their dispensing on microplate wells, and then a rinsing step of the chip before starting a new cycle with the next pDNA. Indeed, these technologies are readily available and just need to be combined [129].
Methods combining microfluidic electroporation and DNA arraying seem at that time more difficult to be widely used. Indeed, many skills are necessary to prepare the functional chip: design of microfluidic device, micro arraying of the DNA, and even a micromanipulation platform to mount the complete functional chip [116]. This and the cost of the required material will limit its use in the scientific community.
Chemical-based transfection is readily available and represents the methods that reached the highest throughputs. The reverse protocol is the preferred mode with the use of lipids or cationic polymers and achieved a throughput of several thousand independent points [104]. A major limitation is that transfection occurs after a suspension step when cells are seeded. The use of the same approaches but in a microarray manner, also showed HT being possible to perform transfection on adherent differentiated cells. However, in this case, the use of microfluidics and their inconvenients impair its wide use.
Biological approaches also reached HT. Viral transduction is the most powerful tool to transfer DNA. However, biosafety concerns, and furthermore difficulties to produce viruses in arrays format avoid its wide use. CPP-based delivery is of great potential and a more important use should be expected in the next decade with the advance of our knowledge in this field.
In order to deliver an easy way to perform transfection even by novices, a fully automated transfection protocol was developed using a tipless nano-acoustic dispenser device [101]. Users just have to indicate amounts of DNA and transfection reagent to be delivered in each well using a custom spreadsheet and prepare the requested source plate. The device-controlled software performs the tedious dispensing from the source plate to destination one, based on the spreadsheet [105]. The method could be applicable to any chemical reagents and even to CPP-based approaches. This approach could also be performed in forward mode then allowing adherent differentiated cells transfection. Newer versions of the device allow 1536-wells plates as the source and can now dispense in 3456-wells plates. It then becomes possible to regroup the human ORFeome collection plasmids on less than 15 sources plates, and their transfer to about seven 3456-wells plates only. The method allows preloading of the plates and long-term storage before cell dispensing. However, the cost of the dispenser is extremely huge and still impairs its use. The future end of the patented technologies protection, expected in 2025–2030, should induce a price drop due to competitors’ and wider the use of such an approach.
The authors thank the National Institute of Health and Medical Research (INSERM) for its financial support for the publication of this chapter.
The booming global businesses have largely facilitated the cross-border flow of goods, but meanwhile are threatened by the dramatically increased intellectual property (IP) crimes nowadays. According to the study by Organization for Economic Cooperation and Development (OECD), the value of counterfeit and pirated products is amounted to USD 464 billion in 2019, equal to 2.5% of world trade and more than half of the total value is carried by containerships between countries [1, 2]. The illicit trade hits company profits and nation tax revenue and endangers public health when pharmaceuticals and medical equipment are involved. For these reasons, advanced technologies that combat fake products demand prompt development to ensure reliable flow of goods while maintain its convenience.
Anti-counterfeiting idea was early raised by Philadelphia printer Benjamin Franklin in the 1700s [3], at that time colonies in North America were troubled by the circulation of counterfeit bills. Franklin deliberately misspelled Pennsylvania in the printed bills to baffle less-literate criminals. Meanwhile, he engraved the fine detail of copper on the leaf vein at the back of each bill, making these bills hard to be reproduced by counterfeiters. The unique copper engraving created by blocky lead printer has been regarded as a prototype for contemporary anti-counterfeiting patterning technologies. Since the 1950s, the development of holograms [4, 5, 6, 7], ink printing [8, 9, 10, 11], and exquisite laser engraving [12, 13, 14] have offered practical solutions to protect the market from malicious third parties.
Halide perovskites as an emerging family of semiconductor materials have achieved notable success in photovoltaics and other optoelectronics over the past decade [15, 16, 17, 18, 19, 20]. The intriguing photophysical property of perovskites, such as widely tunable bandgaps [21, 22, 23, 24, 25, 26], high photoluminescence quantum yield (PLQY) [27, 28, 29], and narrow emission width [30, 31, 32], are making them promising candidates for fabricating luminescent security tags. Meanwhile, the solution/ink processability of perovskites imparts them feasibility with a variety of printing technologies, enabling high-throughput generation of customized labels with enhanced encoding capacity and lowered processing cost [33, 34, 35].
Here, we give a retrospect to the recent advances of halide perovskite-based materials for anti-counterfeiting applications. Low-dimensional perovskites and double perovskites that are structural analogs to three-dimensional (3D) ones as well as other perovskite-like materials are included in the discussion. We summarize the patterning techniques that can lead to precise control of tag fabrication at high dim either flat surface or closed space. The luminescent security tags of perovskites are categorized by different encryption principles, with detailed phase transformation or compositional variation of materials being provided for each chromic case. Integration of luminescent properties that gives rise to multimodal anti-counterfeiting is discussed in respect of goods being strictly confidential. We then survey the special optical readout of security tags that is enabled by the exciton relaxation behavior and carrier dynamic of perovskites.
Taking advantage of the high PLQY of halide perovskites, security information in a luminescent tag can be easily and rapidly identified by the human eye or spectrum. The excitation-dependent emission of perovskites can also be tuned from the monochromatic to broadband white light [36, 37, 38], giving an added complexity to the optical readout of tags. Combined with versatile encryption and decryption strategies, the security level of an individual tag can be enhanced multidimensionally and output in a simplified digital form [39]. The anti-counterfeiting mechanism of security tags during the flow of goods is illustrated in Figure 1, where the authentication is implemented by the communication between preloaded database and third parties.
Anti-counterfeiting mechanism of security tags during the flow of goods.
Perovskite mineral (calcium titanium oxide, CaTiO3) was discovered in the Ural Mountains by German mineralogist Gustav Rose in 1839 [40]. The crystal structure of perovskite oxide was not determined by X-ray diffraction until nearly a century later [41] and was proved to comprise three fundamental phases, i.e. cubic, tetragonal, and orthorhombic based on the rigid 3D lattice. Halide perovskites share the similar crystal structure to perovskite oxide, of which the compounds were first synthesized in the late nineteenth century by H. L. Wells [42]. Typically, 3D perovskites (defined by a chemical formula of ABX3, where A is a monovalent cation, B is a divalent cation, and X is a halide anion) have direct bandgaps that can be widely tuned by altering the composition of A- and B-site cations and halide anions [21, 24, 43, 44]. Besides, 3D perovskites normally feature low exciton binding energy (
Two-dimensional (2D) perovskites feature corner-sharing metal-halide octahedra intercalated by the bulky cations. Emission spectra of 2D perovskites can be structurally correlated with the interlayer spacing, quantum well (QW) thickness, and its distribution [45, 46]. Strong electron-photon coupling that originated from the deformable lattice was previously demonstrated for some 2D perovskite single crystals, which introduces permanent trap states [47]. The self-trapped excitons (STEs) were later revealed to be a type of transient defect driven by the electron-photon coupling and will contribute to the broadband emission of 2D perovskites [48, 49]. Further lowering the dimensionality of 2D perovskites leads to one-dimensional (1D) and zero-dimensional (0D) perovskites whose octahedra are shared by edge or face. STEs can also be responsible for the broadband emission of these materials with large Stokes shift [50, 51, 52, 53]. The white light or dual−/multiband emissions under different excitations are favorable for those luminescent tags that demand a high security level.
Double perovskites are defined by a chemical formula of A2BB’X6, where B is a monovalent cation and B′ is a trivalent cation and feature a rock salt arrangement of BX6 and B’X6 octahedra. In addition, A2B(IV)X6 compounds are also grouped as double perovskites because of their vacancy-ordered structure [54, 55]. The phase-pure double perovskites usually have room-temperature (RT) indirect bandgaps and exhibit band-to-band or downshifting emissions that can be strongly influenced by the specific metal dopants [55, 56, 57, 58, 59]. The in-depth reason was ascribed to lattice distortion since metal dopants will basically affect the length and angle of B − X − B′ bonds and hence change the electronic wave function coupling of metal cations [60].
Halide perovskites possess a high compatibility with printing techniques, since both the precursor solution and synthesized colloidal nanocrystals (NCs) can serve as inks. Using CsPbX3:Mn2+ (X = Cl, Br, I) NCs inks, Wang et al. [34] previously reported the fabrication of various patterns by screen, inkjet, and roll-to-roll printing techniques on flexible substrate (e.g. paper, polyethylene terephthalate, and banknotes). The patterns showed fluorescence as response to 254-nm and 365-nm ultraviolet (UV) light, and the CsPbBr3:Mn2+-based on maintained bright fluorescence after continuous UV irradiation for 60 days. Shi et al. [61] demonstrated an
Nanoscale 3D printing technique was recently reported to fabricate perovskite nanopixels with programmed vertical height, location, and emission characteristics [35], which overcomes the low-resolution problem of conventional printing techniques. The authors of this study used femtoliter meniscus to guide the out-of-plane growth of MAPbX3 (X = Cl, Br, I) crystals from precursor solution, enabling ultrahigh integration density of red, green, and blue (RGB) nanopixel arrays with spacing of ~5 μm while maintaining its lateral resolution (Figure 2a). Numbers can be encoded for each discrete height of nanopixels and thus adds an additional level for encryption. Electrohydrodynamic (EHD) printing as another advanced printing technique was also reported to fabricate high-resolution CsPbX3 (X = Cl, Br, I) dot arrays with full-color display (Figure 2b) [62]. The size of a single dot was precisely controlled by the frequency and peak values of pulse voltage for precursor solution, and a minimum size of 5 μm can be achieved.
(a) Schematic illustration of 3D printing of perovskite nanopixels. (b) Schematic illustration of EHD printing technique for perovskite patterning. (c) Representative laser processing system for perovskite patterning. Reprinted with permission from ref. [
Laser beam was previously used to trigger the ultrafast crystallization of perovskite for both patterning and photovoltaic applications [64]. Figure 2c shows a typical laser processing system for perovskite patterning. Without any heat treatment, Zhang et al. [63] demonstrated the fabrication of CsPbBr3/CsPb2Br5-polymer nanocomposites fluorescent pattern by 532-nm femtosecond laser irradiation. Localized crystallization of perovskite was observed in the irradiated pathway, which was accompanied by the laser-induced polymerization of γ-butyrolactone solvent. The width of perovskite line was lowered down to 1.2 μm, and both the crystal quality and luminescent intensity can be fine-tuned by the power and moving speed of laser beam. In addition, laser engraving was introduced to directly create patterns on CsPbBr3 microplates [65]. The hidden security information provides a guidance for encryption on a miniaturized pattern.
Most recently, Sun et al. [66] reported the use of 3D lithography technique to fabricated separated CsPbX3 (X = Cl, Br, I) NCs in glass matrix. The strong thermal accumulation at the laser-irradiated region of borophosphate glass leads to local pressure and temperature above the liquidus of materials, which induces liquid nanophase separation of glass and perovskite. By tailoring the parameters of pulse duration, repetition rate, pulse energy, and irradiation time, the emission color of pattern was tuned from blue to red under 405-nm excitation. Perovskite NCs in glass matrix exhibited notable phase stability against long-term UV irradiation, organic solution, and high temperature. The patterns were used for both 3D multicolor and dynamic holographic displays, showing huge potential for stereoscopic optical storage and authentication. Accordingly, we provide an overall assessment of existing printing and laser processing techniques for perovskite security tags in Table 1.
Approach | Technique | Dimensionality | Advantage | Disadvantage |
---|---|---|---|---|
Printing | Handwriting [67] | 2D | Easy fabrication, low processing cost | Low-resolution display |
Screen, inkjet, and roll-to-roll printing [34, 61] | 2D | High-throughput fabrication, large-area display | Only available for liquid precursors | |
Electrohydrodynamic printing [62] | 2D | High-resolution display | Conductive substrate required | |
Meniscus-guided printing [35] | 3D | Multidimensional display | Delicate mechanical control of pipet | |
Laser processing | Laser annealing [63, 64] | 2D | Ultrafast fabrication, high-resolution display | Heavy crystallization impact from laser beam |
Laser engraving [33, 65] | 2D | High-resolution display | Flat pattern required | |
Lithography [66] | 3D | Holographic display, high encoding capacity | High-energy laser source required, sophisticated optical paths and machines |
Technical assessment of patterning methods.
With the assistance of advanced patterning techniques, the intriguing luminescent properties found on perovskites can be transformed into security information for encryption and decryption of tags. Normally, these tags are invisible under visible light but can emit light under UV, visible, or near-infrared (NIR) excitations. In this section, we provide an overview of encryption principle of perovskite security tags, including pattern, thermochromism, solvatochromism, photochromism, and multimodal luminescence. Other optical readout, such as long-lived emission (afterglow) phenomenon and carrier lifetime gating, are discussed as special encryption methods for delicate authentication of goods. Figure 3 shows the representative cases of encryption principles being reported over the past few years.
Timeline of pioneering works with new encryption principles being reported for perovskite security tags.
Shape design of a pattern is a fundamental approach to encode the security data relative to the complexity of contours. Printing or laser processing techniques have been developed to create customized pattern shapes whose resolution now reach a few micropixels or below. Lin et al. [33] raised the concept of clonable shape, while unclonable texture for anti-counterfeiting tags is based on CsPbBr3 patterns. A large amount of patterns that grown on laser-engraved lyophilic 1
The vertical height of a single perovskite pixel can be also encoded as specific numbers [35], which is regarded as a complementary encryption strategy to lateral shape design of a pattern (Figure 4a and b). 3D confocal PL imaging was applied to recognize the height variation of perovskite pixels with the height interval of 5 μm. The height values were further converted into binary information matrix for digitalized decryption. As we have mentioned in Section 2.2, the pattern design at three dimensionalities enabled by 3D lithography technique allows more complex encryption on a security tag (Figure 4c–e) [66]. Random 3D luminescent patterns can therefore be spatially and temporally identified, offering an innovative platform for smart authentication of goods.
(a) Tilt-view SEM image of as-printed perovskite nanopixel arrays. (b) Multicolor display of perovskite nanopixel arrays with different halide components under UV light. (c) Multicolor pattern with CsPbClxBr3 − x nanophases in glass under UV light. (d) 3D microhelix arrays of CsPbClxBr3 − x under UV light. (e) Dynamic holographic display of as-patterned “ZJUUSST” characters under 532-nm light. Reprinted with permission from ref. [
Halide perovskites, especially organic–inorganic hybrid ones, feature considerably large thermal expansion coefficients [68, 69]. The thermochromic property of perovskites was first observed in thin film due to the phase transition between transparent hydrated phase (MA4PbI6·2H2O) and dark perovskite phase (MAPbI3) [70]. This phenomenon can be reversible by exposing perovskite film to ambient moisture at RT or heating condition at 60°C repeatably and was explored as the switchable photovoltaic performance for perovskite solar cells. The discoloration mechanism was recently developed for smart window applications based on hydrated MAPbClxI3 − x [71]. Similarly, Lin et al. [72] demonstrated the reversible thermochromic property of CsPbBrxI3 − x film coupled with dynamic transition of RT non-perovskite phase and high-temperature perovskite phase, which is also switched by the moisture and thermal annealing.
Above cases show the thermochromic phenomena of perovskites in the presence of moisture but may not be applicable to anti-counterfeiting tags that are fully encapsulated. Taking advantage of the inverse temperature crystallization (ITC) of hybrid perovskites, Bastiani et al. [73] reported the chromatic inks with wide color variation that depend on the halide constituent of perovskite precipitate. The RT yellow inks turned to orange, red, and black when temperature reached 60°C, 90°C, and 120°C, corresponding to the extrapolated absorption edges of MAPbBr2.7I0.3 at 597 nm, MAPbBr2.4I0.6 at 615 nm, and MAPbBr1.8I1.2 at 651 nm, respectively. The thermochromic behavior of perovskite inks showed consecutive cycling between RT and 60°C for several times.
The reversible thermochromic phenomena was also observed in diphasic perovskite material (CsPbBr3/Cs4PbBr6) wrapped by silica nanosphere [74]. The strong RT PL emission (at 525 nm) of composited patterns gradually decreased when temperature was elevated and almost disappeared at 150°C. Temperature-dependent PL spectra revealed the relatively low activation energy (
Solvatochromism refers to chromic behavior of materials as response to water or other organic solvents. As we mentioned in Section 2.3.2, hybrid perovskites feature hydrochromism due to the formation of hydrated or non-perovskite phases in moisture atmosphere [70, 72]. Reversibly decomposition-induced hydrochromism was recently reported for CsPbBr3 NCs confined in mesoporous silica nanospheres (MSNs) [78]. Orthorhombic CsPbBr3 will decompose into nonluminescent tetragonal CsPb2Br5 and CsBr in the presence of water, and the dissolved CsBr component can be confined in MSNs. As a result, the green emission pattern turned to dark in moisture condition and recovered when water was removed (Figure 5a). Similar hydrochromic mechanism was also reported for CsPbBr3/Cs4PbBr6 nanocomposites, which maintained about half of its initial PL intensity after 10 wetting-drying cycles [80]. Cs3Cu2I5 as lead-free perovskite-like material was recently exploited for hydrochromism-based encryption and decryption of security tags [81, 82, 83]. Water functions as a switch of phase transition between blue emission Cs3Cu2I5 and yellow emission CsCu2I3 under UV excitation. Combined with water-resistant polymethyl methacrylate (PMMA) coating layer, moreover, the microarray patterns can be tailored for dual-color emission toward various shapes and characters in moisture atmosphere [82].
(a) Reversible hydrochromism of CsPbBr3 pattern under 365-nm UV light and the corresponding phase transformation. (b) Reversible DMF-induced solvatochromism of InCl6(C4H10SN)4·Cl:Sb3+ pattern under 365-nm UV light and the corresponding phase transformation. Reprinted with permission from refs. [
Besides water, methanol (MeOH) was previously demonstrated capable to trigger the solvatochromism of MAPbBr3 NCs that are converted from lead-based metal–organic framework (MOF) [84]. The authors of this study found that MeOH impregnation can remove the organic perovskite species while leave lead ions in MOF matrix. The green emission of pattern under UV excitation therefore quickly quenched after impregnation but can be recovered by loading MABr solution (10 mg mL−1 in
Solvatochromism can also be induced by new phase formation where solvent molecules are incorporated into perovskite lattice [79]. The 0D InCl6(C4H10SN)4·Cl:Sb3+ showed red-shifted emission peak from 550 nm to 580 nm and 600 nm when being exposed to ethanol (EtOH) and
Photochromic property has been found in a variety of organics and organic–metal complexes in the case of light-mediated configuration change of molecules [87]. By anchoring the diarylethene (DAE) derivative onto CsPbBr3 QDs surface, Mokhtar et al. [88] observed the reversible photoswitchable luminescence of QDs-DAE hybrids. The open-ring isomer of DAE underwent cyclization under UV light and quickly turned off the green emission of printed pattern, while the green emission can be switched on again by exposing the pattern to visible light for DAE cycloreversion (Figure 6a). Similar photochromic behavior was reported for DAE derivative whose triethoxysilane (TEOS) moiety is altered by alkyl amine [90]. Following this strategy, a majority of photochromic molecules may be introduced as the surfactant to achieve the photochromism of perovskite QDs/NCs.
(a) Photoswitchable cyclization and cycloreversion of DAE surfactant and the resultant photochromism of pattern based on CsPbBr3-DAE hybrids. (b) UV irradiation-induced reversible halide exchange at CsPbCl1.5Br1.5/MYE interface and the photochromism of QR code patterned by CsPbCl1.5Br1.5/MYE composites. Reprinted with permission from refs. [
Photochromism also occurs under the circumstance of photoinduced compositional variation of perovskites. The emission color of CsPbCl1.5Br1.5 NCs that confined in macroporous Y2O3:Eu3+ (MYE) changed from red to green under continuous UV irradiation, which was explained by the halide migration between perovskite NCs and MYE matrix [89]. The small
The bandgap of perovskites is structurally dependent on the QW thickness; in this view, photochromism can be achieved in dimensionality-mixed perovskites whose QW thickness and distribution are self-adapted to light stimulus. The emission behavior of layered FAn + 2PbnBr3n + 2 (FA = formamidinium) was recently studied with respect to its structural transformation under light irradiation [92]. The authors of this study demonstrated the UV damage to perovskite that can convert wide-bandgap 2D phase to narrow-bandgap 3D phase. Accordingly, perovskite film showed emission color changed from blue to green as response to the elongated irradiation time. The metastable 2D phase can meanwhile be transformed back by dark storage, showing reversible photochromism that is applicable for anti-counterfeiting patterns.
Unlike unidirectional authentication methods, multimodal luminescence of perovskites allows the encryption and decryption to be conducted through multiple excited sources. Xu et al. [74] first demonstrated the triple-modal anti-counterfeiting of CsPbBr3@Cs4PbBr6/SiO2 composites in 2017, since the as-patterned codes showed reversible and switchable luminescence to heating, UV, and NIR irradiation. In addition, the dual-color emission of green and red of MAPbBr3@Eu-MOF composites was reported under 365-nm and 254-nm UV lamp [93], respectively, where the red emission under 254-nm excitation primarily comes from the photon upconversion (UC) of Eu-MOF species (Figure 7a and b). Solvatochromism was also observed for the composites, and the written pattern on paper showed reversible green emission via water and MABr treatment. Notably, the UC luminescent component of perovskites can be further tuned by rational doping of lanthanides [94].
(a) The dependence of PL spectra of MAPbBr3@Eu-MOF composites on the UV excitation wavelength. (b) Hydrochromism of “USTB” characters based on MAPbBr3@Eu-MOF composites and the MABr-induced recovery under 254-nm and 365-nm UV light. (c) Photographs of Cs2Ag0.6Na0.4InCl6:Yb3+/Er3+/Bi3+ (RE-1) under different excitations. (d) XEL, DS-PL, and UC-PL spectra of RE-1. (e) Photographs of RE-1 pattern under visible and 365 nm UV light. Reprinted with permission from refs. [
Overcoming the limited response range of conventional perovskite materials, the excitation source of Yb3+/Er3+/Bi3+ co-doped Cs2Ag0.6Na0.4InCl6 double perovskite was reported to be extended to X-ray, as a complementary to UV and NIR [58]. Bi3+ ions were demonstrated to reduce the structural disorder, promote the exciton localization, and lead to strong Jahn-Teller effect that would benefit both UC and X-ray excited luminescence (XEL) (Figure 7c and d). The as-synthesized double-perovskite single crystals were ground and dispersed in organic solvent for ink printing, and the patterns showed exceptional luminescent stability in thermal heating (up to 400°C), moisture, and high-dosage radiation conditions (Figure 7e). The combination of X-ray excited luminescence (XEL), downshifting (DS), UC luminescence, and other routine encryption methods enhance the confidential level of tags considerably, which offers a reliable solution for customized authentication of high-value products.
Some special optical readout of perovskites can be transformed into security information for anti-counterfeiting applications. Here, we exemplify the encryption principles of patterns based on afterglow phenomenon and carrier lifetime gating. The RT afterglow of perovskites was first reported for 2D PEA2PbCl4 (PEA = phenylethylammonium) perovskite doped with 1,8-naphthalimide (NI) spacers [95]. The as-printed pattern on paper showed UV-excited white emission in nitrogen atmosphere that comprises blue fluorescence from perovskite and yellow phosphorescence from NI organic cations. After UV light off, however, the blue fluorescence (PLQY: 25.6%) quenched quickly, while the yellow phosphorescence (PLQY: 56.1%) can maintain for a few seconds. This property caused the yellow afterglow of pattern that can be identified by both spectrum and human eye. Wei et al. [96] recently found the RT greenish afterglow of 0D BAPPIn1.996Sb0.004Cl10 (BAPP = C10H28N4) perovskite-like material after UV light off, where the relaxation of excitons from BAPP organic cations were demonstrated to be responsible for the afterglow (Figure 8a–d). For CsPbBr3 NCs doped by lanthanide ions (Ln3+), the persistent time of afterglow is even up to 1800 s [98]. In addition, X-ray-induced afterglow was also reported for 0D Cs4EuX6 (X = Br, I) perovskite single crystals, despite the case did not involve anti-counterfeiting applications [99].
(a) Molecular configuration of BAPP4+ cation and crystal structure of BAPPIn2Cl10. (b–d) Photographs of BAPPIn1.996Sb0.004Cl10 pattern under visible light, 365-nm UV light, and 365-nm UV light off (afterglow), respectively. (e) FLIM image and (f) time-correlated single-photon counting fluorescence lifetime imaging (TCSPC-FLI) image of tag patterned by CsPbBr3 and {en}FAPbBr3 NCs inks. (g) Fast-lifetime histograms of as-patterned inks and (h) binarization of lifetime for QR code generation. Reprinted with permission from refs. [
The carrier lifetime of perovskites is influenced by a variety of factors, among which the composition of perovskite can be the deterministic one. The EHD-printed security tags were reported to be encrypted based on the different carrier lifetime of CsPbBr3 and hollowed {en}FAPbBr3 NCs, which can then be decrypted by either fluorescence-lifetime imaging microscopy (FLIM) or time-of-flight fluorescence-lifetime imaging (ToF-FLI) (Figure 8e–h) [97]. These two imaging techniques enabled machine-readable lifetime of QR code that cannot be readily decoded by routine methods. Moreover, the system is highly reconfigurable due to the compositional versatility of perovskite NCs. The enhancement and Purcell factors of CsPbClxBr3 − x QDs that coupled to plasmonic silver cavity were also extracted for the encryption of QR code, where the factors are defined by the relationship among excitation efficiency, light extraction efficiency, quantum efficiency, and radiative rate [100].
We hereby briefly discuss the current challenges encountered by perovskite fluorescent tags prior to their real-world applications, including the potential overuse of toxic lead, the poor durability, and many clonable functions that can be easily reproduced by counterfeiters. Possible solutions are also provided with respect to each challenging case.
Lead’s toxicity has been widely recognized due to its damage to the nervous system of biological individuals. Therefore, lead-based wastes are now under strict control in many developed countries. Despite perovskite security tags made by lead compounds feature many intriguing fluorescent properties, they can be highly risky when adhere to daily goods and cause potential lead leakage. Alternatively, more environmental-friendly perovskites (e.g. tin-, antimony-, bismuth-, and copper-based) can be developed to replace lead-based ones while maintaining the bright luminescence and high processability of tags [55, 57, 58, 79, 82, 83, 96].
The phase stability of halide perovskites, especially 3D ones, can be susceptible to environmental perturbations and hence fail to work during long term or repeated authentication. Lowering down the dimensionality of perovskites as well as composite strategies enable more robust perovskite phase, yet the stability of fluorescent tags can hardly rival the simple-patterned tags (e.g. QR code). Advanced sealing techniques alleviate this problem by isolating perovskites from environment; however, they are limit for those tags that need direct exposure to atmosphere, chemicals, or solvents. Inert matrix has been demonstrated to enhance the durability of both common and special perovskite fluorescent tags. Beside glass, silica, and polymers [66, 82, 101], other durable matrix materials remain to be exploited.
Single-mode perovskite fluorescent tags work as response to certain stimulus, making their functions clonable by commercial phosphors or other functionalized luminescent materials. A safer communication between users and server database requires physically unclonable functions (PUFs) that generated by irregular encryption and decryption methods. In this view, multimodal anti-counterfeiting that combines two or more encoding and decoding pathways (see Section 2.3) is prompt to be developed for highly confidential security tags. In addition, authentication based on the digital readout of sophisticated machines can also fulfill the demands of PUFs [97, 100].
Increasingly rich encryption principles have been exploited for halide perovskite-based security tags owning to their intriguing luminescent properties as response to a wide range of stimuli. Apart from the existing cases, the mechanochromism upon mechanical stress as well as the magnetochromism under altered magnetic field can be studied for perovskites with the aim of further enriching the diversity of authentication methods [102, 103]. Perovskite memristors as a new rising technology was also demonstrated to deliver switching electronic signals relative to the charged defects and halide motions inside the materials, providing an additional solution toward the design of PUF system [104]. All these unique optical and digital readout may overcome the limit of conventional clonable tags such as QR codes, watermarks, and raised print.
Future development of perovskite security tags is supposed to follow the taxonomy of predominant PUFs, including high encoding capacity, tunable security level, logically/physically reconfigurable functions, and switchable access between private and public. Based on the rational screening strategy of perovskite materials, micro- and nanoscale patterning techniques allow these functions to be multidimensionally integrated in a minimized tag, making security information more robust against third parties. Halide perovskites are bound to play a more important role in anti-counterfeiting arena and contribute to future smart flow of goods in a more fair and orderly global market.
YH thanks the support by National Ten Thousand Talent Program for Young Topnotch Talent.
The authors declare no conflict of interest.
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Relationships",subtitle:null,isOpenForSubmission:!1,hash:"ebf41f4d17c75010eb3294cc8cac3d47",slug:"interpersonal-relationships",bookSignature:"Martha Peaslee Levine",coverURL:"https://cdn.intechopen.com/books/images_new/7827.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"186919",title:"Dr.",name:"Martha",middleName:null,surname:"Peaslee Levine",slug:"martha-peaslee-levine",fullName:"Martha Peaslee Levine"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10908",title:"Advances in Decision Making",subtitle:null,isOpenForSubmission:!1,hash:"126486f7f91e18e2e3539a32c38be7b1",slug:"advances-in-decision-making",bookSignature:"Fausto Pedro García Márquez",coverURL:"https://cdn.intechopen.com/books/images_new/10908.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"22844",title:"Prof.",name:"Fausto Pedro",middleName:null,surname:"García Márquez",slug:"fausto-pedro-garcia-marquez",fullName:"Fausto Pedro García Márquez"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10669",title:"Corrosion",subtitle:"Fundamentals and Protection Mechanisms",isOpenForSubmission:!1,hash:"4a76d54f8a40fc2e7002a8d13fd617c1",slug:"corrosion-fundamentals-and-protection-mechanisms",bookSignature:"Fahmina Zafar, Anujit Ghosal and Eram Sharmin",coverURL:"https://cdn.intechopen.com/books/images_new/10669.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"89672",title:"Dr.",name:"Fahmina",middleName:null,surname:"Zafar",slug:"fahmina-zafar",fullName:"Fahmina Zafar"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10677",title:"Advanced Topics of Topology",subtitle:null,isOpenForSubmission:!1,hash:"bf964c52f9e653fac20a7fcab58070e5",slug:"advanced-topics-of-topology",bookSignature:"Francisco Bulnes",coverURL:"https://cdn.intechopen.com/books/images_new/10677.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"92918",title:"Dr.",name:"Francisco",middleName:null,surname:"Bulnes",slug:"francisco-bulnes",fullName:"Francisco Bulnes"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"11195",title:"Recent Advances in Biometrics",subtitle:null,isOpenForSubmission:!1,hash:"2d32e33e0f499cb5241734bb75dd2a83",slug:"recent-advances-in-biometrics",bookSignature:"Muhammad Sarfraz",coverURL:"https://cdn.intechopen.com/books/images_new/11195.jpg",editedByType:"Edited by",publishedDate:"July 27th 2022",editors:[{id:"215610",title:"Prof.",name:"Muhammad",middleName:null,surname:"Sarfraz",slug:"muhammad-sarfraz",fullName:"Muhammad Sarfraz"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},subject:{topic:{id:"1161",title:"Andrology",slug:"andrology",parent:{id:"204",title:"Urology",slug:"urology"},numberOfBooks:6,numberOfSeries:0,numberOfAuthorsAndEditors:179,numberOfWosCitations:40,numberOfCrossrefCitations:43,numberOfDimensionsCitations:109,videoUrl:null,fallbackUrl:null,description:null},booksByTopicFilter:{topicId:"1161",sort:"-publishedDate",limit:12,offset:0},booksByTopicCollection:[{type:"book",id:"10724",title:"Male Reproductive Anatomy",subtitle:null,isOpenForSubmission:!1,hash:"a3fdda3194735da4287e9ea193beb07e",slug:"male-reproductive-anatomy",bookSignature:"Wei Wu",coverURL:"https://cdn.intechopen.com/books/images_new/10724.jpg",editedByType:"Edited by",editors:[{id:"178661",title:"Dr.",name:"Wei",middleName:null,surname:"Wu",slug:"wei-wu",fullName:"Wei Wu"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7985",title:"Circumcision and the Community",subtitle:null,isOpenForSubmission:!1,hash:"023cc135aeeae6d2ea8cfc01ab3f4dc7",slug:"circumcision-and-the-community",bookSignature:"Ahmad Zaghal and Nishat Rahman",coverURL:"https://cdn.intechopen.com/books/images_new/7985.jpg",editedByType:"Edited by",editors:[{id:"240621",title:"Dr.",name:"Ahmad",middleName:null,surname:"Zaghal",slug:"ahmad-zaghal",fullName:"Ahmad Zaghal"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7931",title:"Male Reproductive Health",subtitle:null,isOpenForSubmission:!1,hash:"5754baea5de6a634c66bae12a33d52d9",slug:"male-reproductive-health",bookSignature:"Wei Wu, Francesco Ziglioli and Umberto Maestroni",coverURL:"https://cdn.intechopen.com/books/images_new/7931.jpg",editedByType:"Edited by",editors:[{id:"178661",title:"Dr.",name:"Wei",middleName:null,surname:"Wu",slug:"wei-wu",fullName:"Wei Wu"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6079",title:"Spermatozoa",subtitle:"Facts and Perspectives",isOpenForSubmission:!1,hash:"2d4488814a6ea68efcd3544209c9e4d2",slug:"spermatozoa-facts-and-perspectives",bookSignature:"Rosaria Meccariello and Rosanna Chianese",coverURL:"https://cdn.intechopen.com/books/images_new/6079.jpg",editedByType:"Edited by",editors:[{id:"143980",title:"Prof.",name:"Rosaria",middleName:null,surname:"Meccariello",slug:"rosaria-meccariello",fullName:"Rosaria Meccariello"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"972",title:"Male Infertility",subtitle:null,isOpenForSubmission:!1,hash:"92b68c49e083613bc65d3db92f6aca22",slug:"male-infertility",bookSignature:"Anu Bashamboo and Kenneth David McElreavey",coverURL:"https://cdn.intechopen.com/books/images_new/972.jpg",editedByType:"Edited by",editors:[{id:"87226",title:"Dr.",name:"Anu",middleName:null,surname:"Bashamboo",slug:"anu-bashamboo",fullName:"Anu Bashamboo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"686",title:"Erectile Dysfunction",subtitle:"Disease-Associated Mechanisms and Novel Insights into Therapy",isOpenForSubmission:!1,hash:"c5caa41eb9d576f7765dfcb06a6df94c",slug:"erectile-dysfunction-disease-associated-mechanisms-and-novel-insights-into-therapy",bookSignature:"Kenia Pedrosa Nunes",coverURL:"https://cdn.intechopen.com/books/images_new/686.jpg",editedByType:"Edited by",editors:[{id:"71405",title:"Dr.",name:"Kenia",middleName:"Pedrosa",surname:"Nunes",slug:"kenia-nunes",fullName:"Kenia Nunes"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],booksByTopicTotal:6,seriesByTopicCollection:[],seriesByTopicTotal:0,mostCitedChapters:[{id:"30215",doi:"10.5772/39088",title:"\ufeffMechanisms in Erectile Function and Dysfunction: An Overview",slug:"mechanisms-in-erectile-function-and-dysfunction-an-overview",totalDownloads:7483,totalCrossrefCites:1,totalDimensionsCites:19,abstract:null,book:{id:"686",slug:"erectile-dysfunction-disease-associated-mechanisms-and-novel-insights-into-therapy",title:"Erectile Dysfunction",fullTitle:"Erectile Dysfunction - Disease-Associated Mechanisms and Novel Insights into Therapy"},signatures:"Kenia Pedrosa Nunes and R. Clinton Webb",authors:[{id:"71405",title:"Dr.",name:"Kenia",middleName:"Pedrosa",surname:"Nunes",slug:"kenia-nunes",fullName:"Kenia Nunes"},{id:"134106",title:"Dr.",name:"R. Clinton",middleName:null,surname:"Webb",slug:"r.-clinton-webb",fullName:"R. Clinton Webb"}]},{id:"59074",doi:"10.5772/intechopen.73231",title:"The Role of Human Semen as an Early and Reliable Tool of Environmental Impact Assessment on Human Health",slug:"the-role-of-human-semen-as-an-early-and-reliable-tool-of-environmental-impact-assessment-on-human-he",totalDownloads:1544,totalCrossrefCites:7,totalDimensionsCites:11,abstract:"Several studies have shown a dramatic reduction of semen quality in many industrialized countries and infertility is becoming a public health top priority, whose incidence is associated to late-onset adult diseases, especially cancer, shorter life expectancy and trans-generational effects. The male reproductive system is particularly sensitive to a broad variety of reproductive and developmental toxicants, including many environmental pollutants and recent studies suggest that human semen is an early and sensitive environmental and health marker. A set of semen biomarkers is described for reproductive health effects in relation to environmental exposure, where human semen seems to be an early and sensitive source of biomarkers than blood to monitor high environmental pressure on human health. Environmental health should consider reproductive health and development, from intrauterine life to childhood and puberty: these are both vulnerable targets and high-value protection goals, inasmuch as they represent the future of our societies. Hence, biomarkers of reproductive health should be exploited as early signals of environmental pressure and increased risk of adverse chronic health effects so that the use of “human seminal model” might be the main objective to be considered in the agenda of public prevention policies for early detection and innovative programs of health surveillance in environmental risk areas.",book:{id:"6079",slug:"spermatozoa-facts-and-perspectives",title:"Spermatozoa",fullTitle:"Spermatozoa - Facts and Perspectives"},signatures:"Luigi Montano, Paolo Bergamo, Maria Grazia Andreassi and\nStefano Lorenzetti",authors:[{id:"206180",title:"Dr.",name:"Luigi",middleName:null,surname:"Montano",slug:"luigi-montano",fullName:"Luigi Montano"},{id:"222782",title:"Dr.",name:"Paolo",middleName:null,surname:"Bergamo",slug:"paolo-bergamo",fullName:"Paolo Bergamo"},{id:"222783",title:"Dr.",name:"Maria Grazia",middleName:null,surname:"Andreassi",slug:"maria-grazia-andreassi",fullName:"Maria Grazia Andreassi"},{id:"222784",title:"Dr.",name:"Stefano",middleName:null,surname:"Lorenzetti",slug:"stefano-lorenzetti",fullName:"Stefano Lorenzetti"}]},{id:"36145",doi:"10.5772/32617",title:"Apoptosis, ROS and Calcium Signaling in Human Spermatozoa: Relationship to Infertility",slug:"apoptosis-ros-and-calcium-signaling-in-human-spermatozoa-relationship-to-infertility",totalDownloads:3022,totalCrossrefCites:6,totalDimensionsCites:11,abstract:null,book:{id:"972",slug:"male-infertility",title:"Male Infertility",fullTitle:"Male Infertility"},signatures:"Ignacio Bejarano, Javier Espino, Sergio D. Paredes, Águeda Ortiz, Graciela Lozano, José Antonio Pariente, Ana B. Rodríguez",authors:[{id:"92130",title:"Dr.",name:"Ignacio",middleName:null,surname:"Bejarano",slug:"ignacio-bejarano",fullName:"Ignacio Bejarano"},{id:"98354",title:"MSc.",name:"Javier",middleName:null,surname:"Espino",slug:"javier-espino",fullName:"Javier Espino"},{id:"98356",title:"Dr.",name:"Sergio",middleName:null,surname:"Paredes",slug:"sergio-paredes",fullName:"Sergio Paredes"},{id:"98362",title:"MSc.",name:"Águeda",middleName:null,surname:"Ortiz",slug:"agueda-ortiz",fullName:"Águeda Ortiz"},{id:"98374",title:"Dr.",name:"Graciela",middleName:null,surname:"Lozano",slug:"graciela-lozano",fullName:"Graciela Lozano"},{id:"98375",title:"Prof.",name:"José Antonio",middleName:null,surname:"Pariente",slug:"jose-antonio-pariente",fullName:"José Antonio Pariente"},{id:"98376",title:"Prof.",name:"Ana Beatriz",middleName:null,surname:"Rodríguez",slug:"ana-beatriz-rodriguez",fullName:"Ana Beatriz Rodríguez"}]},{id:"57417",doi:"10.5772/intechopen.70793",title:"Physiological and Pathological Roles of Free Radicals in Male Reproduction",slug:"physiological-and-pathological-roles-of-free-radicals-in-male-reproduction",totalDownloads:1416,totalCrossrefCites:4,totalDimensionsCites:7,abstract:"Oxidative stress (OS) is a condition caused by an imbalance between reactive oxygen species (ROS) overgeneration and decreased antioxidant defense mechanisms in the cell. OS has become a prominent factor in male reproductive dysfunction as ROS cause damage to sperm DNA, lipids and proteins, alterations to critical sperm structures and signaling pathways, leading to a decreased sperm activity and fertilizing capacity. At the same time, small amounts of ROS play vital roles in events leading to sperm maturation and acquisition of functional activity, which is why a proper oxidative balance is of paramount importance for a proper male fertility. Understanding the physiological and pathological roles of ROS in male reproduction has become an essential pillar of modern andrology; however, numerous questions related to the controversial behavior of ROS in male reproductive cells and tissues still remain unanswered. This chapter aims to summarize current evidence available on the relationships between free radicals, antioxidants and male reproduction and to trigger more scientific interest, particularly with respect to the design of efficient strategies to diagnose or treat male sub- or infertility associated with OS.",book:{id:"6079",slug:"spermatozoa-facts-and-perspectives",title:"Spermatozoa",fullTitle:"Spermatozoa - Facts and Perspectives"},signatures:"Eva Tvrdá, Peter Massanyi and Norbert Lukáč",authors:[{id:"204993",title:"Dr.",name:"Eva",middleName:null,surname:"Tvrdá",slug:"eva-tvrda",fullName:"Eva Tvrdá"},{id:"206075",title:"Prof.",name:"Norbert",middleName:null,surname:"Lukáč",slug:"norbert-lukac",fullName:"Norbert Lukáč"},{id:"220755",title:"Prof.",name:"Peter",middleName:null,surname:"Massanyi",slug:"peter-massanyi",fullName:"Peter Massanyi"}]},{id:"67581",doi:"10.5772/intechopen.86808",title:"Sertoli Cell Phagocytosis: An Essential Event for Spermatogenesis",slug:"sertoli-cell-phagocytosis-an-essential-event-for-spermatogenesis",totalDownloads:1059,totalCrossrefCites:1,totalDimensionsCites:6,abstract:"During spermatogenesis, most male germ cells undergo apoptosis, and the cytoplasmic portions of the elongating spermatids are shed as residual bodies (RB). Both apoptotic germ cells (AGC) and RB must be phagocytosed by Sertoli cells, which are essential to maintain testicular homeostasis for normal spermatogenesis. The phagocytosis of AGC and RB by Sertoli cells confers various meanings, including elimination of apoptotic components, removal of autoantigens, and the recycle of degenerated substrates as an energy source. Sertoli cell phagocytosis can be regulated by various mechanisms. The impairment of Sertoli cell phagocytosis may disrupt tissue homeostasis in the testis, thereby impairing to testicular function and spermatogenesis. This chapter discusses the mechanisms underlying phagocytic removal of AGC and RB by Sertoli cells and the consequences of this biological event for spermatogenesis and male fertility.",book:{id:"7931",slug:"male-reproductive-health",title:"Male Reproductive Health",fullTitle:"Male Reproductive Health"},signatures:"Fei Wang and Daishu Han",authors:[{id:"295978",title:"Dr.",name:"Daishu",middleName:null,surname:"Han",slug:"daishu-han",fullName:"Daishu Han"},{id:"303373",title:"Dr.",name:"Fei",middleName:null,surname:"Wang",slug:"fei-wang",fullName:"Fei Wang"}]}],mostDownloadedChaptersLast30Days:[{id:"57404",title:"Assessment of Human Sperm Cells Morphological Parameters",slug:"assessment-of-human-sperm-cells-morphological-parameters",totalDownloads:1363,totalCrossrefCites:0,totalDimensionsCites:1,abstract:"The quality of spermatozoa has a direct influence on the fertilization and developmental competence of embryos. The aim of this work was to review the methods of spermatozoa morphology assessment, features of the normal spermatozoa and the reasons of their several abnormalities. Three methods can be used for the evaluation of spermatozoa morphology in the in vitro fertilization (IVF) laboratory: (1) light microscopy of stained spermatozoa, (2) motile sperm organelle morphology examination (MSOME) and (3) polarized light microscopy. The analysis of spermatozoa morphology includes the assessment of head, neck, midpiece and tail. Morphologically abnormal spermatozoa are categorized into subgroups according to the defects of the head, neck, midpiece and/or tail. Before IVF and intracytoplasmic sperm injection (ICSI), the quality of spermatozoa must be estimated exactly, because this has the high influence on embryo development. Therefore the analysis of the morphological parameters of spermatozoa using the light microscopy, MSOME, in combination with precise head birefringence detection using the polarized microscopy, could give the best fertilization rate and embryo quality after IVF and ICSI.",book:{id:"6079",slug:"spermatozoa-facts-and-perspectives",title:"Spermatozoa",fullTitle:"Spermatozoa - Facts and Perspectives"},signatures:"Kristina Lasiene",authors:[{id:"206099",title:"Dr.",name:"Kristina",middleName:null,surname:"Lasiene",slug:"kristina-lasiene",fullName:"Kristina Lasiene"}]},{id:"79167",title:"The Concept of Male Reproductive Anatomy",slug:"the-concept-of-male-reproductive-anatomy",totalDownloads:232,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"The human reproductive system is made up of the primary and secondary organs, which helps to enhances reproduction. The male reproductive system is designed to produce male gametes and convey them to the female reproductive tract through the use of supportive fluids and testosterone synthesis. The paired testis (site of testosterone and sperm generation), scrotum (compartment for testis localisation), epididymis, vas deferens, seminal vesicles, prostate gland, bulbourethral gland, ejaculatory duct, urethra, and penis are the parts of the male reproductive system. The auxiliary organs aid in the maturation and transportation of sperm. Semen is made up of sperm and the secretions of the seminal vesicles, prostate, and bulbourethral glands (the ejaculate). Ejaculate is delivered to the female reproduc¬tive tract by the penis and urethra. The anatomy, embryology and functions of the male reproductive system are discussed in this chapter.",book:{id:"10724",slug:"male-reproductive-anatomy",title:"Male Reproductive Anatomy",fullTitle:"Male Reproductive Anatomy"},signatures:"Oyovwi Mega Obukohwo, Nwangwa Eze Kingsley, Rotu Arientare Rume and Emojevwe Victor",authors:[{id:"348295",title:"Dr.",name:"Oyovwi",middleName:null,surname:"Mega Obukohwo",slug:"oyovwi-mega-obukohwo",fullName:"Oyovwi Mega Obukohwo"},{id:"421235",title:"Prof.",name:"Nwangwa",middleName:null,surname:"Eze Kingsley",slug:"nwangwa-eze-kingsley",fullName:"Nwangwa Eze Kingsley"},{id:"428844",title:"Dr.",name:"Rotu",middleName:null,surname:"Arientare Rume",slug:"rotu-arientare-rume",fullName:"Rotu Arientare Rume"}]},{id:"78480",title:"Methods of Sperm Selection for In-Vitro Fertilization",slug:"methods-of-sperm-selection-for-in-vitro-fertilization-1",totalDownloads:262,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"50–60% of infertility cases are as a result of male infertility and infertile men semen sample is characterize with poor motility, abnormal morphology, low sperm concentration, azoospermic and increased levels of sperm DNA damage. As a result of this heterogeneity of the ejaculate, sperm selection has become a necessary step to carry out prior to in vitro fertilization. Furthermore, the choice of sperm cell selection techniques depend on sperm concentration and sperm biology and the recovery of highly functional sperm cell population depend on the combination of more than one technique in some cases. The regular sperm cell selection methods in ART laboratory are swim up, density gradient, simple wash and other advanced and emerging sperm selection techniques which include hyaluronic acid mediated sperm binding, Zeta potential, hypoosmotic swelling test, magnetic activated cell sorting and microfluidic separation of sperm cells. The various methods have its own advantages and disadvantages which may be applicable to the individual need of infertile men and its effect on ART outcome.",book:{id:"10724",slug:"male-reproductive-anatomy",title:"Male Reproductive Anatomy",fullTitle:"Male Reproductive Anatomy"},signatures:"Abimibola Nanna",authors:[{id:"349382",title:"M.Sc.",name:"Abimibola",middleName:null,surname:"Nanna",slug:"abimibola-nanna",fullName:"Abimibola Nanna"}]},{id:"78100",title:"Epigenetics in Male Infertility",slug:"epigenetics-in-male-infertility",totalDownloads:223,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"Male infertility is a complex medical condition, in which epigenetic factors play an important role. Epigenetics has recently gained significant scientific attention since it has added a new dimension to genomic and proteomic research. As a mechanism for maintaining genomic integrity and controlling gene expression, epigenetic modifications hold a great promise in capturing the subtle, yet very important, regulatory elements that might drive normal and abnormal sperm functions. The sperm’s epigenome is known to be marked by constant changing over spermatogenesis, which is highly susceptible to be influenced by a wide spectrum of environmental stimuli. Recently, epigenetic aberrations have been recognized as one of the causes of idiopathic male infertility. Recent advances in technology have enabled humans to study epigenetics role in male infertility.",book:{id:"10724",slug:"male-reproductive-anatomy",title:"Male Reproductive Anatomy",fullTitle:"Male Reproductive Anatomy"},signatures:"Hayfa H. Hassani, Rakad M. Kh AL-Jumaily and Fadhel M. Lafta",authors:[{id:"349295",title:"Prof.",name:"Hayfa H.",middleName:null,surname:"Hassani",slug:"hayfa-h.-hassani",fullName:"Hayfa H. Hassani"},{id:"350281",title:"Prof.",name:"Fadhel M.",middleName:null,surname:"Lafta",slug:"fadhel-m.-lafta",fullName:"Fadhel M. Lafta"},{id:"352246",title:"Dr.",name:"Rakad",middleName:null,surname:"M. Kh AL-Jumaily",slug:"rakad-m.-kh-al-jumaily",fullName:"Rakad M. Kh AL-Jumaily"}]},{id:"77407",title:"Positional Relationships among Male Reproductive Organs in Insects",slug:"positional-relationships-among-male-reproductive-organs-in-insects",totalDownloads:192,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"The location, morphology and function of male internal reproductive organs in insects have been extensively studied, but the relative positioning of those organs is less understood. Position and morphology of the testis, vas deferens, seminal vesicle, accessory gland and ejaculatory duct determine the migration or ejaculation of sperm and other substances. In species where the testis is connected with the seminal vesicle directly or the seminal vesicle is lacking, males usually store complete sperm in the testis and thus can use them immediately for mating. In contrast, the testis of lepidopteran insects is separated from the duplex (sperm storage organ) via the vas deferens, and the sperm are not mature, requiring morphological development in the vas deferens. Here, we discuss the significance of various positional relationships of male reproductive organs and how this relates to their morphology and function with a focus on sperm.",book:{id:"10724",slug:"male-reproductive-anatomy",title:"Male Reproductive Anatomy",fullTitle:"Male Reproductive Anatomy"},signatures:"Satoshi Hiroyoshi and Gadi V.P. Reddy",authors:[{id:"348309",title:"Dr.",name:"Satoshi",middleName:null,surname:"Hiroyoshi",slug:"satoshi-hiroyoshi",fullName:"Satoshi Hiroyoshi"},{id:"348547",title:"Dr.",name:"Gadi",middleName:null,surname:"V.P. 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He has an excellent track record in the herpesvirus field, and his group is engaged in clinical research in the field of Epstein-Barr virus diseases. He is the editor of the online Encyclopedia of Environment and he coordinates the Universal Health Coverage education program for the BioHealth Computing Schools of the European Institute of Science.",institutionString:null,institution:{name:"Grenoble Alpes University",country:{name:"France"}}},{id:"131400",title:"Prof.",name:"Alfonso J.",middleName:null,surname:"Rodriguez-Morales",slug:"alfonso-j.-rodriguez-morales",fullName:"Alfonso J. Rodriguez-Morales",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/131400/images/system/131400.png",biography:"Dr. Rodriguez-Morales is an expert in tropical and emerging diseases, particularly zoonotic and vector-borne diseases (especially arboviral diseases). He is the president of the Travel Medicine Committee of the Pan-American Infectious Diseases Association (API), as well as the president of the Colombian Association of Infectious Diseases (ACIN). He is a member of the Committee on Tropical Medicine, Zoonoses, and Travel Medicine of ACIN. He is a vice-president of the Latin American Society for Travel Medicine (SLAMVI) and a Member of the Council of the International Society for Infectious Diseases (ISID). Since 2014, he has been recognized as a Senior Researcher, at the Ministry of Science of Colombia. He is a professor at the Faculty of Medicine of the Fundacion Universitaria Autonoma de las Americas, in Pereira, Risaralda, Colombia. He is an External Professor, Master in Research on Tropical Medicine and International Health, Universitat de Barcelona, Spain. He is also a professor at the Master in Clinical Epidemiology and Biostatistics, Universidad Científica del Sur, Lima, Peru. In 2021 he has been awarded the “Raul Isturiz Award” Medal of the API. Also, in 2021, he was awarded with the “Jose Felix Patiño” Asclepius Staff Medal of the Colombian Medical College, due to his scientific contributions to COVID-19 during the pandemic. He is currently the Editor in Chief of the journal Travel Medicine and Infectious Diseases. His Scopus H index is 47 (Google Scholar H index, 68).",institutionString:"Institución Universitaria Visión de las Américas, Colombia",institution:null},{id:"332819",title:"Dr.",name:"Chukwudi Michael",middleName:"Michael",surname:"Egbuche",slug:"chukwudi-michael-egbuche",fullName:"Chukwudi Michael Egbuche",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/332819/images/14624_n.jpg",biography:"I an Dr. Chukwudi Michael Egbuche. I am a Senior Lecturer in the Department of Parasitology and Entomology, Nnamdi Azikiwe University, Awka.",institutionString:null,institution:{name:"Nnamdi Azikiwe University",country:{name:"Nigeria"}}},{id:"284232",title:"Mr.",name:"Nikunj",middleName:"U",surname:"Tandel",slug:"nikunj-tandel",fullName:"Nikunj Tandel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/284232/images/8275_n.jpg",biography:'Mr. Nikunj Tandel has completed his Master\'s degree in Biotechnology from VIT University, India in the year of 2012. He is having 8 years of research experience especially in the field of malaria epidemiology, immunology, and nanoparticle-based drug delivery system against the infectious diseases, autoimmune disorders and cancer. He has worked for the NIH funded-International Center of Excellence in Malaria Research project "Center for the study of complex malaria in India (CSCMi)" in collaboration with New York University. The preliminary objectives of the study are to understand and develop the evidence-based tools and interventions for the control and prevention of malaria in different sites of the INDIA. Alongside, with the help of next-generation genomics study, the team has studied the antimalarial drug resistance in India. Further, he has extended his research in the development of Humanized mice for the study of liver-stage malaria and identification of molecular marker(s) for the Artemisinin resistance. At present, his research focuses on understanding the role of B cells in the activation of CD8+ T cells in malaria. Received the CSIR-SRF (Senior Research Fellow) award-2018, FIMSA (Federation of Immunological Societies of Asia-Oceania) Travel Bursary award to attend the IUIS-IIS-FIMSA Immunology course-2019',institutionString:"Nirma University",institution:{name:"Nirma University",country:{name:"India"}}},{id:"334383",title:"Ph.D.",name:"Simone",middleName:"Ulrich",surname:"Ulrich Picoli",slug:"simone-ulrich-picoli",fullName:"Simone Ulrich Picoli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/334383/images/15919_n.jpg",biography:"Graduated in Pharmacy from Universidade Luterana do Brasil (1999), Master in Agricultural and Environmental Microbiology from Federal University of Rio Grande do Sul (2002), Specialization in Clinical Microbiology from Universidade de São Paulo, USP (2007) and PhD in Sciences in Gastroenterology and Hepatology (2012). She is currently an Adjunct Professor at Feevale University in Medicine and Biomedicine courses and a permanent professor of the Academic Master\\'s Degree in Virology. She has experience in the field of Microbiology, with an emphasis on Bacteriology, working mainly on the following topics: bacteriophages, bacterial resistance, clinical microbiology and food microbiology.",institutionString:null,institution:{name:"Universidade Feevale",country:{name:"Brazil"}}},{id:"229220",title:"Dr.",name:"Amjad",middleName:"Islam",surname:"Aqib",slug:"amjad-aqib",fullName:"Amjad Aqib",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/229220/images/system/229220.png",biography:"Dr. Amjad Islam Aqib obtained a DVM and MSc (Hons) from University of Agriculture Faisalabad (UAF), Pakistan, and a PhD from the University of Veterinary and Animal Sciences Lahore, Pakistan. 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He received his medical degree from the Sousse Faculty of Medicine at Sousse, University of Sousse, Tunisia. He completed his surgical residency in General Surgery at the University Hospital Farhat Hached of Sousse and was a member of the Unit of Liver Transplantation in the University of Rennes, France. He then worked in the Department of Surgery at the Sahloul University Hospital in Sousse. Professor Derbel is presently working at the Clinique les Oliviers, Sousse, Tunisia. His hospital activities are mostly concerned with laparoscopic, colorectal, pancreatic, hepatobiliary, and gastric surgery. He is also very interested in hernia surgery and performs ventral hernia repairs and inguinal hernia repairs. He has been a member of the GREPA and Tunisian Hernia Society (THS). During his residency, he managed patients suffering from diabetic foot, and he was very interested in this pathology. For this reason, he decided to coordinate a book project dealing with the diabetic foot. Professor Derbel has published many articles in journals and collaborates intensively with IntechOpen Access Publisher as an editor.",institutionString:"Clinique les Oliviers",institution:null},{id:"300144",title:"Dr.",name:"Meriem",middleName:null,surname:"Braiki",slug:"meriem-braiki",fullName:"Meriem Braiki",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/300144/images/system/300144.jpg",biography:"Dr. Meriem Braiki is a specialist in pediatric surgeon from Tunisia. She was born in 1985. She received her medical degree from the University of Medicine at Sousse, Tunisia. She achieved her surgical residency training periods in Pediatric Surgery departments at University Hospitals in Monastir, Tunis and France.\r\nShe is currently working at the Pediatric surgery department, Sidi Bouzid Hospital, Tunisia. Her hospital activities are mostly concerned with laparoscopic, parietal, urological and digestive surgery. She has published several articles in diffrent journals.",institutionString:"Sidi Bouzid Regional Hospital",institution:null},{id:"229481",title:"Dr.",name:"Erika M.",middleName:"Martins",surname:"de Carvalho",slug:"erika-m.-de-carvalho",fullName:"Erika M. de Carvalho",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/229481/images/6397_n.jpg",biography:null,institutionString:null,institution:{name:"Oswaldo Cruz Foundation",country:{name:"Brazil"}}},{id:"186537",title:"Prof.",name:"Tonay",middleName:null,surname:"Inceboz",slug:"tonay-inceboz",fullName:"Tonay Inceboz",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/186537/images/system/186537.jfif",biography:"I was graduated from Ege University of Medical Faculty (Turkey) in 1988 and completed his Med. PhD degree in Medical Parasitology at the same university. I became an Associate Professor in 2008 and Professor in 2014. I am currently working as a Professor at the Department of Medical Parasitology at Dokuz Eylul University, Izmir, Turkey.\n\nI have given many lectures, presentations in different academic meetings. I have more than 60 articles in peer-reviewed journals, 18 book chapters, 1 book editorship.\n\nMy research interests are Echinococcus granulosus, Echinococcus multilocularis (diagnosis, life cycle, in vitro and in vivo cultivation), and Trichomonas vaginalis (diagnosis, PCR, and in vitro cultivation).",institutionString:"Dokuz Eylül University",institution:{name:"Dokuz Eylül University",country:{name:"Turkey"}}},{id:"71812",title:"Prof.",name:"Hanem Fathy",middleName:"Fathy",surname:"Khater",slug:"hanem-fathy-khater",fullName:"Hanem Fathy Khater",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/71812/images/1167_n.jpg",biography:"Prof. Khater is a Professor of Parasitology at Benha University, Egypt. She studied for her doctoral degree, at the Department of Entomology, College of Agriculture, Food and Natural Resources, University of Missouri, Columbia, USA. She has completed her Ph.D. degrees in Parasitology in Egypt, from where she got the award for “the best scientific Ph.D. dissertation”. She worked at the School of Biological Sciences, Bristol, England, the UK in controlling insects of medical and veterinary importance as a grant from Newton Mosharafa, the British Council. Her research is focused on searching of pesticides against mosquitoes, house flies, lice, green bottle fly, camel nasal botfly, soft and hard ticks, mites, and the diamondback moth as well as control of several parasites using safe and natural materials to avoid drug resistances and environmental contamination.",institutionString:null,institution:{name:"Banha University",country:{name:"Egypt"}}},{id:"99780",title:"Prof.",name:"Omolade",middleName:"Olayinka",surname:"Okwa",slug:"omolade-okwa",fullName:"Omolade Okwa",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/99780/images/system/99780.jpg",biography:"Omolade Olayinka Okwa is presently a Professor of Parasitology at Lagos State University, Nigeria. She has a PhD in Parasitology (1997), an MSc in Cellular Parasitology (1992), and a BSc (Hons) Zoology (1990) all from the University of Ibadan, Nigeria. She teaches parasitology at the undergraduate and postgraduate levels. She was a recipient of a Commonwealth fellowship supported by British Council tenable at the Centre for Entomology and Parasitology (CAEP), Keele University, United Kingdom between 2004 and 2005. She was awarded an Honorary Visiting Research Fellow at the same university from 2005 to 2007. \nShe has been an external examiner to the Department of Veterinary Microbiology and Parasitology, University of Ibadan, MSc programme between 2010 and 2012. She is a member of the Nigerian Society of Experimental Biology (NISEB), Parasitology and Public Health Society of Nigeria (PPSN), Science Association of Nigeria (SAN), Zoological Society of Nigeria (ZSN), and is Vice Chairperson of the Organisation of Women in Science (OWSG), LASU chapter. She served as Head of Department of Zoology and Environmental Biology, Lagos State University from 2007 to 2010 and 2014 to 2016. She is a reviewer for several local and international journals such as Unilag Journal of Science, Libyan Journal of Medicine, Journal of Medicine and Medical Sciences, and Annual Research and Review in Science. \nShe has authored 45 scientific research publications in local and international journals, 8 scientific reviews, 4 books, and 3 book chapters, which includes the books “Malaria Parasites” and “Malaria” which are IntechOpen access publications.",institutionString:"Lagos State University",institution:{name:"Lagos State University",country:{name:"Nigeria"}}},{id:"273100",title:"Dr.",name:"Vijay",middleName:null,surname:"Gayam",slug:"vijay-gayam",fullName:"Vijay Gayam",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/273100/images/system/273100.jpeg",biography:"Dr. Vijay Bhaskar Reddy Gayam is currently practicing as an internist at Interfaith Medical Center in Brooklyn, New York, USA. He is also a Clinical Assistant Professor at the SUNY Downstate University Hospital and Adjunct Professor of Medicine at the American University of Antigua. He is a holder of an M.B.B.S. degree bestowed to him by Osmania Medical College and received his M.D. at Interfaith Medical Center. His career goals thus far have heavily focused on direct patient care, medical education, and clinical research. He currently serves in two leadership capacities; Assistant Program Director of Medicine at Interfaith Medical Center and as a Councilor for the American\r\nFederation for Medical Research. As a true academician and researcher, he has more than 50 papers indexed in international peer-reviewed journals. He has also presented numerous papers in multiple national and international scientific conferences. His areas of research interest include general internal medicine, gastroenterology and hepatology. He serves as an editor, editorial board member and reviewer for multiple international journals. His research on Hepatitis C has been very successful and has led to multiple research awards, including the 'Equity in Prevention and Treatment Award” from the New York Department of Health Viral Hepatitis Symposium (2018) and the 'Presidential Poster Award” awarded to him by the American College of Gastroenterology (2018). He was also awarded 'Outstanding Clinician in General Medicine” by Venus International Foundation for his extensive research expertise and services, perform over and above the standard expected in the advancement of healthcare, patient safety and quality of care.",institutionString:"Interfaith Medical Center",institution:{name:"Interfaith Medical Center",country:{name:"United States of America"}}},{id:"93517",title:"Dr.",name:"Clement",middleName:"Adebajo",surname:"Meseko",slug:"clement-meseko",fullName:"Clement Meseko",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/93517/images/system/93517.jpg",biography:"Dr. Clement Meseko obtained DVM and PhD degree in Veterinary Medicine and Virology respectively. He has worked for over 20 years in both private and public sectors including the academia, contributing to knowledge and control of infectious disease. Through the application of epidemiological skill, classical and molecular virological skills, he investigates viruses of economic and public health importance for the mitigation of the negative impact on people, animal and the environment in the context of Onehealth. \r\nDr. Meseko’s field experience on animal and zoonotic diseases and pathogen dynamics at the human-animal interface over the years shaped his carrier in research and scientific inquiries. He has been part of the investigation of Highly Pathogenic Avian Influenza incursions in sub Saharan Africa and monitors swine Influenza (Pandemic influenza Virus) agro-ecology and potential for interspecies transmission. He has authored and reviewed a number of journal articles and book chapters.",institutionString:"National Veterinary Research Institute",institution:{name:"National Veterinary Research Institute",country:{name:"Nigeria"}}},{id:"158026",title:"Prof.",name:"Shailendra K.",middleName:null,surname:"Saxena",slug:"shailendra-k.-saxena",fullName:"Shailendra K. Saxena",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRET3QAO/Profile_Picture_2022-05-10T10:10:26.jpeg",biography:"Professor Dr. Shailendra K. Saxena is a vice dean and professor at King George's Medical University, Lucknow, India. His research interests involve understanding the molecular mechanisms of host defense during human viral infections and developing new predictive, preventive, and therapeutic strategies for them using Japanese encephalitis virus (JEV), HIV, and emerging viruses as a model via stem cell and cell culture technologies. His research work has been published in various high-impact factor journals (Science, PNAS, Nature Medicine) with a high number of citations. He has received many awards and honors in India and abroad including various Young Scientist Awards, BBSRC India Partnering Award, and Dr. JC Bose National Award of Department of Biotechnology, Min. of Science and Technology, Govt. of India. Dr. Saxena is a fellow of various international societies/academies including the Royal College of Pathologists, United Kingdom; Royal Society of Medicine, London; Royal Society of Biology, United Kingdom; Royal Society of Chemistry, London; and Academy of Translational Medicine Professionals, Austria. He was named a Global Leader in Science by The Scientist. He is also an international opinion leader/expert in vaccination for Japanese encephalitis by IPIC (UK).",institutionString:"King George's Medical University",institution:{name:"King George's Medical University",country:{name:"India"}}},{id:"94928",title:"Dr.",name:"Takuo",middleName:null,surname:"Mizukami",slug:"takuo-mizukami",fullName:"Takuo Mizukami",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/94928/images/6402_n.jpg",biography:null,institutionString:null,institution:{name:"National Institute of Infectious Diseases",country:{name:"Japan"}}},{id:"233433",title:"Dr.",name:"Yulia",middleName:null,surname:"Desheva",slug:"yulia-desheva",fullName:"Yulia Desheva",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/233433/images/system/233433.png",biography:"Dr. Yulia Desheva is a leading researcher at the Institute of Experimental Medicine, St. Petersburg, Russia. She is a professor in the Stomatology Faculty, St. Petersburg State University. She has expertise in the development and evaluation of a wide range of live mucosal vaccines against influenza and bacterial complications. Her research interests include immunity against influenza and COVID-19 and the development of immunization schemes for high-risk individuals.",institutionString:'Federal State Budgetary Scientific Institution "Institute of Experimental Medicine"',institution:null},{id:"238958",title:"Mr.",name:"Atamjit",middleName:null,surname:"Singh",slug:"atamjit-singh",fullName:"Atamjit Singh",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/238958/images/6575_n.jpg",biography:null,institutionString:null,institution:null},{id:"252058",title:"M.Sc.",name:"Juan",middleName:null,surname:"Sulca",slug:"juan-sulca",fullName:"Juan Sulca",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/252058/images/12834_n.jpg",biography:null,institutionString:null,institution:null},{id:"191392",title:"Dr.",name:"Marimuthu",middleName:null,surname:"Govindarajan",slug:"marimuthu-govindarajan",fullName:"Marimuthu Govindarajan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/191392/images/5828_n.jpg",biography:"Dr. M. Govindarajan completed his BSc degree in Zoology at Government Arts College (Autonomous), Kumbakonam, and MSc, MPhil, and PhD degrees at Annamalai University, Annamalai Nagar, Tamil Nadu, India. He is serving as an assistant professor at the Department of Zoology, Annamalai University. His research interests include isolation, identification, and characterization of biologically active molecules from plants and microbes. He has identified more than 20 pure compounds with high mosquitocidal activity and also conducted high-quality research on photochemistry and nanosynthesis. He has published more than 150 studies in journals with impact factor and 2 books in Lambert Academic Publishing, Germany. He serves as an editorial board member in various national and international scientific journals.",institutionString:null,institution:null},{id:"274660",title:"Dr.",name:"Damodar",middleName:null,surname:"Paudel",slug:"damodar-paudel",fullName:"Damodar Paudel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/274660/images/8176_n.jpg",biography:"I am DrDamodar Paudel,currently working as consultant Physician in Nepal police Hospital.",institutionString:null,institution:null},{id:"241562",title:"Dr.",name:"Melvin",middleName:null,surname:"Sanicas",slug:"melvin-sanicas",fullName:"Melvin Sanicas",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/241562/images/6699_n.jpg",biography:null,institutionString:null,institution:null},{id:"322007",title:"Dr.",name:"Maria Elizbeth",middleName:null,surname:"Alvarez-Sánchez",slug:"maria-elizbeth-alvarez-sanchez",fullName:"Maria Elizbeth Alvarez-Sánchez",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universidad Autónoma de la Ciudad de México",country:{name:"Mexico"}}},{id:"337443",title:"Dr.",name:"Juan",middleName:null,surname:"A. Gonzalez-Sanchez",slug:"juan-a.-gonzalez-sanchez",fullName:"Juan A. Gonzalez-Sanchez",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Puerto Rico System",country:{name:"United States of America"}}},{id:"337446",title:"Dr.",name:"Maria",middleName:null,surname:"Zavala-Colon",slug:"maria-zavala-colon",fullName:"Maria Zavala-Colon",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Puerto Rico, Medical Sciences Campus",country:{name:"United States of America"}}},{id:"338856",title:"Mrs.",name:"Nur Alvira",middleName:null,surname:"Pascawati",slug:"nur-alvira-pascawati",fullName:"Nur Alvira Pascawati",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universitas Respati Yogyakarta",country:{name:"Indonesia"}}}]}},subseries:{item:{id:"3",type:"subseries",title:"Bacterial Infectious Diseases",keywords:"Antibiotics, Biofilm, Antibiotic Resistance, Host-microbiota Relationship, Treatment, Diagnostic Tools",scope:"