Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by FSW and FSSW.
\r\n\tThis book will describe the self-assembly of materials and supramolecular chemistry design principles for a broad spectrum of materials, including bio-inspired amphiphiles, metal oxides, metal nanoparticles, and organic-inorganic hybrid materials. It will provide fundamental concepts of self-assembly design approaches and supramolecular chemistry principles for research ideas in nanotechnology applications. The book will focus on three main themes, which include: the self-assembly and supramolecular chemistry of amphiplies by coordination programming, the supramolecular structures and devices of inorganic materials, and the assembly-disassembly of organic-inorganic hybrid materials. The contributing chapters will be written by leading scientists in their field, with the hope that this book will provide a foundation on supramolecular chemistry principles to students and active researchers who are interested in nanoscience and nanoengineering fields.
",isbn:"978-1-83969-702-9",printIsbn:"978-1-83969-701-2",pdfIsbn:"978-1-83969-703-6",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,isNomenclature:!1,hash:"e9cc643ae0a219e91e445a1e61b33a22",bookSignature:"Prof. Hemali Rathnayake and Dr. Gayani Pathiraja",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11908.jpg",keywords:"Amphiphiles, Artificial Siderophores, Coordination Chemistry, Self-Assembly Design, Supramolecular Structures, Metal Oxides, Metal Particles, 2D Inorganic Materials, Supramolecular Devices, Stimuli-Responsive Materials, Assembly-Disassembly Design, Superstructures",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"April 27th 2022",dateEndSecondStepPublish:"July 1st 2022",dateEndThirdStepPublish:"August 30th 2022",dateEndFourthStepPublish:"November 18th 2022",dateEndFifthStepPublish:"January 17th 2023",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"6 days",secondStepPassed:!0,areRegistrationsClosed:!1,currentStepOfPublishingProcess:3,editedByType:null,kuFlag:!1,biosketch:"Dr. Rathnayake is a pioneering researcher in self-assembly and supramolecular chemistry, with a Ph.D. from the University of Massachusetts Amherst, US. She is an inventor of three innovative technologies, including the Bioinspried Sub-7 nm self-assembled structures for patterning, and holder of multiple registered patents.",coeditorOneBiosketch:"Dr. Gayani Pathiraja is a Postdoctoral Research Scholar at the Joint School of Nanoscience and Nanoengineering (JSNN). She received her Ph.D. in Nanoscience from the University of North Carolina at Greensboro in 2021. Her research interests focus on the crystal growth mechanism and kinetics of metal oxide nanostructure formation via self-assembly.",coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"323782",title:"Prof.",name:"Hemali",middleName:null,surname:"Rathnayake",slug:"hemali-rathnayake",fullName:"Hemali Rathnayake",profilePictureURL:"https://mts.intechopen.com/storage/users/323782/images/system/323782.jpg",biography:"Dr. Hemali Rathnayake, Associate Professor in the Department of Nanoscience at the Joint School of Nanoscience and Nanoengineering, the University of North Carolina at Greensboro, USA, obtained her B.S. in Chemistry from the University of Peradeniya in Sri Lanka. She obtained her Ph.D. from the University of Massachusetts Amherst (UMass), Department of Chemistry in 2007. She was a Postdoctoral research fellow at Polymer Science & Engineering, UMass Amherst. \r\nDr. Rathnayake is a pioneer scientist and a chemist in the field of Nanomaterials Chemistry, with a focus on the interfacial interaction of nanomaterials, molecules, macromolecules, and polymers in homogeneous and heterogeneous media. Her research on the design, synthesis, self-assembly, and application of well-defined superstructures in nanoelectronics, environmental remediation, and sustainable energy has impacted the scientific community with highly rated peer-reviewed journals publications, and more than 80 invited talks to scientific and non-scientific communities including colleges and high schools.",institutionString:"University of North Carolina at Greensboro",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"2",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"University of North Carolina at Greensboro",institutionURL:null,country:{name:"United States of America"}}}],coeditorOne:{id:"427650",title:"Dr.",name:"Gayani",middleName:null,surname:"Pathiraja",slug:"gayani-pathiraja",fullName:"Gayani Pathiraja",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00003CCSN2QAP/Profile_Picture_1644217020559",biography:"Dr. Gayani Pathiraja is a Postdoctoral Research Scholar at the Joint School of Nanoscience and Nanoengineering (JSNN). She received her Ph.D. in Nanoscience from the University of North Carolina at Greensboro (UNCG) in 2021. Her expertise area of focus is investigating the crystal growth mechanism and kinetics of metal oxide nanostructure formation via in-situ self-assembly design principles. \r\nDr. Pathiraja earned her master’s degree in electrochemistry/Environmental Engineering from the University of Peradeniya, Sri Lanka, and her Bachelor’s degree in Materials Science and Technology from Uva Wellassa University, Sri Lanka. Dr. Pathiraja started her academic career as a lecturer at the Department of Engineering Technology, University of Ruhuna, Sri Lanka in 2016. She is a co-author of several peer-reviewed journal publications and a book chapter, and she has presented her work at several regional, international, and national conferences.",institutionString:"University of North Carolina at Greensboro",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"University of North Carolina at Greensboro",institutionURL:null,country:{name:"United States of America"}}},coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"8",title:"Chemistry",slug:"chemistry"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"466998",firstName:"Dragan",lastName:"Miljak",middleName:"Anton",title:"Mr.",imageUrl:"https://mts.intechopen.com/storage/users/466998/images/21564_n.jpg",email:"dragan@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review, to approval and revision, copy-editing and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"314",title:"Regenerative Medicine and Tissue Engineering",subtitle:"Cells and Biomaterials",isOpenForSubmission:!1,hash:"bb67e80e480c86bb8315458012d65686",slug:"regenerative-medicine-and-tissue-engineering-cells-and-biomaterials",bookSignature:"Daniel Eberli",coverURL:"https://cdn.intechopen.com/books/images_new/314.jpg",editedByType:"Edited by",editors:[{id:"6495",title:"Dr.",name:"Daniel",surname:"Eberli",slug:"daniel-eberli",fullName:"Daniel Eberli"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"57",title:"Physics and Applications of Graphene",subtitle:"Experiments",isOpenForSubmission:!1,hash:"0e6622a71cf4f02f45bfdd5691e1189a",slug:"physics-and-applications-of-graphene-experiments",bookSignature:"Sergey Mikhailov",coverURL:"https://cdn.intechopen.com/books/images_new/57.jpg",editedByType:"Edited by",editors:[{id:"16042",title:"Dr.",name:"Sergey",surname:"Mikhailov",slug:"sergey-mikhailov",fullName:"Sergey Mikhailov"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1373",title:"Ionic Liquids",subtitle:"Applications and Perspectives",isOpenForSubmission:!1,hash:"5e9ae5ae9167cde4b344e499a792c41c",slug:"ionic-liquids-applications-and-perspectives",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/1373.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"2270",title:"Fourier Transform",subtitle:"Materials Analysis",isOpenForSubmission:!1,hash:"5e094b066da527193e878e160b4772af",slug:"fourier-transform-materials-analysis",bookSignature:"Salih Mohammed Salih",coverURL:"https://cdn.intechopen.com/books/images_new/2270.jpg",editedByType:"Edited by",editors:[{id:"111691",title:"Dr.Ing.",name:"Salih",surname:"Salih",slug:"salih-salih",fullName:"Salih Salih"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"872",title:"Organic Pollutants Ten Years After the Stockholm Convention",subtitle:"Environmental and Analytical Update",isOpenForSubmission:!1,hash:"f01dc7077e1d23f3d8f5454985cafa0a",slug:"organic-pollutants-ten-years-after-the-stockholm-convention-environmental-and-analytical-update",bookSignature:"Tomasz Puzyn and Aleksandra Mostrag-Szlichtyng",coverURL:"https://cdn.intechopen.com/books/images_new/872.jpg",editedByType:"Edited by",editors:[{id:"84887",title:"Dr.",name:"Tomasz",surname:"Puzyn",slug:"tomasz-puzyn",fullName:"Tomasz Puzyn"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"68643",title:"Self-Fluorescence of Photosynthetic System: A Powerful Tool for Investigation of Microalgal Biological Diversity",doi:"10.5772/intechopen.88785",slug:"self-fluorescence-of-photosynthetic-system-a-powerful-tool-for-investigation-of-microalgal-biologica",body:'\nCyanobacteria are the most ancient photosynthetic microorganisms on Earth. Nowadays, cyanobacteria are one of the most widespread organisms in nature, and the ecological aspect in their investigation is quite valuable. On the other hand, thousands of strains belonging to different species are cultivated in biolaboratories all over the world for different cyanobacterial biotechnological applications such as biofuel cells, food production, pharmaceuticals, fertilizers, etc. [1, 2, 3]. Thus the noninvasive spectroscopic methods are quite requisite for monitoring of physiological state of cyanobacterial cultures and natural communities.
\nIt is well-known that the analysis of self-fluorescence of photosynthetic system is a powerful noninvasive tool for investigation of microalgae in vivo. It reports on the energy transfer and trapping and, thus, reflects the metabolic mechanisms in photosynthetic cells and their photosynthetic efficiency. The detected self-fluorescence finally reflects the diversity in morphological and physiological states of photosynthetic cells [4, 5, 6].
\nSelf-fluorescence originates from excited states that were lost before photochemistry took place and usually represents a small fraction of the excited state decay in a functional photosynthetic complex. Nevertheless, this small fraction can be easily detected by confocal laser scanning microscopy (CLSM). With the confocal fluorescence microscopy, a very small excitation and detection areas can be investigated, so that single cells under non-damage conditions can be studied in vivo. Single-cell detection can provide the information on small peculiarities that is regularly buried in normal ensemble average experiments. This is thus a good way to study the time evolution process and spectroscopic properties of individual cells. Both steady-state and time-resolved fluorescence measurements can be used for probing the organization and functioning of photosynthetic systems by means of CLSM.
\nTill now the best taxonomic differentiation is still obtained using classical inverted microscopy. Unfortunately, this method is time-consuming, human based, and requires appropriate technical skills; this eliminates the possibility of its application for continuous online monitoring. Nearly single-cell flow cytometric analysis, based on light scattering by the cells and fluorescence of the chlorophylls and the phycobilins, can be easily automated, but it is appropriate only for unicellular species and is useless for numerous industrially cultured filamentous strains [7, 8]. The main problem of all chemical methods (e.g., high-performance liquid chromatography (HPLC) [9, 10]) is that during the chemical sample preparation, the most of the information about the peculiarities of individual species is lost and the residual part of the information is not enough for species/strain discrimination inside cyanobacterial genera and is suitable only for the rude differentiation of big classes of phytoplankton. Thus, the analysis of the in vivo fluorescence spectra is the only one noninvasive technique for obtaining qualitative information about the phytoplankton abundance and composition, which is continuously demonstrated by various publications [10, 11, 12, 13, 14, 15, 16, 17]. The relative phytoplankton abundance can be calculated once initial assumptions about the phytoplankton classes are presented and their pigment compositions have been made [7, 12, 13].
\nMaybe the first attempt to use phycoerythrins as chemotaxonomic markers was done by Glazer et al. [18] for red algae in 1982, but until now fluorescence spectra of phycobilins do not appear to be useful at familial, ordinal, and class levels in taxonomic studies. Although the investigation in [18] concerns only purified high-molecular-weight phycoerythrin from red algae, this work clearly demonstrates the possibility of the correct taxonomic analysis on the basis of phycobiliproteins structural differences, which can serve as intrinsical fingerprints for taxons and genera in phytoplankton diversity. Later, the correlation between the distribution of the biliproteins and the genera of Cryptophyceae was discussed in [19]. In 1985, Yentsch and Phinney [20] proposed an ataxonomic technique that utilized the spectral fluorescence signatures of major ocean phytoplankton. Seppälä [16] used spectral fluorescence signals to detect changes in the phytoplankton community. In 2002, Beutler et al. reported a reduced model of the fluorescence from the cyanobacterial photosynthetic apparatus designed for the in situ detection of cyanobacteria and presented a commercially available diveable instrument for online monitoring of phytoplankton structure [11].
\nHowever, the correct classification of cyanobacterial species on the basis of their bulk fluorescence signature is hampered by alterations in pigment composition within one strain, which depends on the physiological state of the culture (community) and environmental conditions [21]. On the other hand, several researchers show that the nutrient and light limitations do not significantly change the initial fluorescence spectra and cannot impede the species discrimination [17, 22].
\nRecent rapid development of confocal microscope functionality initiates new directions in subcellular biology research [23, 24]. Confocal laser scanning microscopes are distinguished by their high spatial and temporal resolution. Modern laser scanning microscopes are unique tools for visualizing cellular structures and analyzing dynamic processes inside single cells. One of the specific fields of CLSM application is the investigation of self-fluorescence of living cells. CLSM single-cell microscopic spectroscopy is undoubtedly the most powerful tool for in vivo investigation of physiological processes in photosynthetic organisms (cyanobacteria, algae, and higher plants). The investigation of self-fluorescence of single living cells reveals the relation between the physiological state and the operational activity of photosynthetic system. A lot of interesting static and dynamic effects can be studied by means of CLSM. The investigation of self-fluorescence gives the information about single-cell processes as well as about the collaboration in cell communities. Changes in spectral characteristics of living photosynthetic cells indicate changes in their physiological state and can be applied for the studies of the results of stress states and external actions [4, 5, 6]. Moreover, the diversity in single-cell self-fluorescence for different species and strains can serve the basis for ataxonomic discrimination of cyanobacterial genera.
\nIn this chapter, a novel ataxonomic approach to differentiation of cyanobacterial cells based on the numerical analysis of in vivo single-cell fluorescence spectra, recorded by means of CLSM, is presented. The differentiation is conducted according to the structure and operation of their photosynthetic apparatus. An optimal set of the parameters is selected, which is sufficient for determination of the taxonomic position of cyanobacteria by means of mathematical statistics. On the basis of the linear discriminant analysis, the obtained spectroscopic data for 23 cyanobacterial strains from CALU collection were analyzed. It was shown that the presented technique allows an accurate differentiation of cyanobacteria up to the species/strain level and enables to distinguish automatically potentially harmful strains. All presented results were obtained using cyanobacterial strains from CALU collection of the Core Facility Center “Centre for Culture Collection of Microorganisms” of the Research Park of St. Petersburg State University as model objects for CLSM studies.
\nAll work on preparing cyanobacteria cultures for this research was carried out at the Core Facility Center “Centre for Culture Collection of Microorganisms” of the Research Park of St. Petersburg State University. In the CALU collection [25] at the core facility center, cyanobacterial strains were maintained in semiliquid agar (0.8%) medium no. 6 after Gromov [26] in test tubes of volume 5–6 mL under cotton plugs. The strains were stored at 14°C under a constant illumination of 2000 lux and were recultivated with a periodicity of 2–3 months.
\nCyanobacteria used in this investigation were grown on liquid medium no. 6. A stock culture was preliminarily prepared, for which it was cultivated in 30 mL of medium and incubated for 2 weeks at room temperature under continuous illumination from fluorescent lamps. To maintain a constant volume, 5 mL of medium were added to the stock culture every 2 weeks. All experiments in this study were conducted with cultures presumably in the logarithmic phase of their growth.
\nIn this work, 23 cyanobacterial strains from CALU collection were used:
Merismopedia sp. CALU 666 punctata Meyen f., Pinar del Rio, Rio de Soroa, Cuba.
Microcystis firma sp. CALU 398 (Breb. et Lenorm) Schmidle, Turkmenbashi Canal, Turkmenistan.
Fluorescent and corresponding transmission photomicrographs, obtained via CLSM, for several strains from CALU collection are presented in Figure 1. In further illustrations only the CALU numbers for corresponding strains will be used for the clarity of the narration.
\nCLSM fluorescent and transmission photomicrographs for eight cyanobacterial strains from CALU collection. The white bar indicates the object scale.
Confocal laser scanning microscopes are distinguished by their high spatial and temporal resolution [23, 24]. Modern laser scanning microscopes are unique tools for visualizing cellular structures and analyzing dynamic processes inside single cells. They exceed classical light microscopes especially in their axial resolution, which enables to acquire optical sections (slices) of a specimen. Apart from simple imaging, confocal laser scanning microscopes are designed for the quantification and analysis of image-coded information. Among other things, they allow easy determination of fluorescence intensities, distances, areas, and their changes over time. New acquisition CLSM tools include the detection of quantitative properties of the emitted light such as spectral signatures and fluorescence lifetimes. The most impressive feature of modern CLSMs is their capability for single-cell microscopic spectroscopy, which allows to obtain spectroscopic information inside single cells and small regions.
\nIn the present investigation, Leica TCS-SP5 was used for the investigation of living cyanobacterial cells. Fluorescence emission spectra of the intact cells were measured at eight excitation wavelengths corresponding to all available laser lines. The excitation wavelengths are 458, 476, 488, 496, and 514 nm are the lines of Ar laser, 405 nm is the line of diode UV laser, and 543 and 633 nm are the lines of HeNe laser. In all presented experiments, laser power settings are as follows: 29% of Ar laser power was reflected onto sample with acousto-optical tunable filter (AOTF), and further power percentage for its laser lines was 30% of 458 nm laser line and 10% for all other lines. 405 nm line of diode UV laser was reflected onto sample with 3%; HeNe laser lines 543 and 633 nm were reflected with 10 and 2%, respectively. An acousto-optical beam splitter (AOBS) was used to transmit sample fluorescence to the detector. Emission spectra between 520 and 785 nm were recorded using the lambda scan function of the “Leica Confocal Software” by sequentially acquiring a series (‘stack’) of 38–45 images, each with a 6 nm fluorescence detection bandwidth and with 6 nm wavelength step. For obtaining fluorescence-intensity information, images of 512 × 512 pixels were collected with a 63× Glycerol immersion lens (Glycerol 80% H2O) with a numeric aperture of 1.3 (objective HCX PL APO 63.0 × 1.30 GLYC 37°C UV) and with additional digital zoom factor 5–9 (depending on a cyanobacterial strain). One pixel corresponds to 53.5 × 53.5 nm. The photomultiplier (PMT) voltages were used in range from 900 to 1100 V. The fluorescence emission images were accompanied with the transmission images (in the parallel channel), collected by a transmission detector with the photomultiplier voltages ranged from 300 to 500 V. For better signal yield, lambda scans were performed with “low speed” setting (400 Hz) in bidirectional scan mode and with a pinhole setting of 1 Airy unit (the inner light circle of the diffraction pattern of a point light source corresponds to a diameter of 102.9 μm with the lens used (see [23]). Regions of interest (ROIs) representing single cells or subcellular regions were used to calculate fluorescence spectra.
\nFor 2D imaging, to raise the sensitivity and contrast, images were recorded at 405 nm excitation wavelength (diod UV laser) and by Leica HyD hybrid detector, which strongly improves contrast in comparison to PMTs. HyD gain was taken as 100 V. The images of 1024 × 1024 and 2048 × 2048 pixels were collected with a 63× Glycerol immersion lens (Glycerol 80% H2O) with a numeric aperture of 1.3 (objective HCX PL APO 63.0 × 1.30 GLYC 37°C UV) and with additional digital zoom factor 10–35. The fluorescence emission images were accompanied with the transmission images (in the parallel channel). The images were recorded with a pinhole setting of 1 Airy unit.
\nIn CLSM applications, the laser light density in the focus point is high. But, generally, it is deposited in short “dwell times” during the laser scanning process. Dwell time and the intervals between the illuminations may influence photodamage and saturation of photosynthetic apparatus of living cells. Thus, since most chromophores bleach under the high laser excitation energies, a bleach test should be performed [27]. It was shown experimentally that especially phycoerythrin (PE) and phycocyanin (PC), as accessory pigments, were very sensitive to photobleaching, while the fluorescence of chlorophyll a (Chl a) and allophycocyanin (APC) remained stable in the intact living cells [27]. During the detection the fluorescence of the main accessory pigments for each cyanobacterial strain should be controlled and the changes in their fluorescence should not exceed 10–20%. In this investigation the power of individual laser lines was chosen according to the photodamage they cause. The repeated spectra were obtained under selected excitation power at a fixed point in a cell to check whether the excitation would affect the cells. It was shown that at the above chosen excitation energies (laser line percentage) the fluorescence spectra did not vary within the experimental error during 10–15 records. When excitation energy was increased, both the height and the center of the bands varied enormously with time because of photodamage or structure breakdown in photosynthetic systems. In the experiments, where several laser lines were involved for the investigation, the first spectrum was recorded again at the end of each series to control the initial state of the cell. It should be pointed out that the whole procedure of fluorescence spectra recording, used in this study, was designed to minimize preparatory manipulation, so as to conduct a noninvasive investigation of small amounts of experimental material and to prevent any damage of living cells.
\nTo exclude unpredictable variations in physiological state of investigated cultures, the fluorescence spectra were taken from the cells of one strain several times, at different days and for various developmental stages of the culture. And it was established that the variations in spectrum shape and intensity among cells of one strain are not considerable.
\nThe main difficulty of the considered discrimination problem resulted from the high nonuniformity of the initial data and different numbers of observation for different strains. A small size of initial dataset as well as the sophisticated nature of the experimental data required a complex preprocessing procedure. The original experimental data represents 307 sets of self-fluorescence spectra obtained from cyanobacterial cells, belonging to 23 different strains. Each observation from a data set is described by a series of seven spectra taken from a single cell by means of CLSM. Each initial spectrum is an array of 38–45 numbers, which correspond to the fluorescence intensities on specific emission frequencies of visible light in the range from 520 to 785 nm. In contrast to the previous investigations, which utilized for classification a full spectrum of the samples [12, 13, 14], we used a set of integral and statistical characteristics, describing the shape of each spectrum. To extract a set of classification parameters from initial data, a computer program has been developed in a mathematics system MATLAB. By means of this program, normalization, interpolation, extrapolation, and smoothing of the raw spectra were carried out, to eliminate the random noise and metering fluctuations. All spectra were reduced to the same scale and size of data array, the first derivative was taken over initial spectra, and the fast Fourier transform (FFT) was performed, to exclude random noise, owing to the low intensity of the exciting and emitting light. The specific values characterizing the shape of obtained curves and the spectral composition of their derivatives were calculated. All selected classification parameters can be divided into three groups: asymmetry and excess, fluorescence emission percentage for individual pigments in four main spectral regions (phycoerythrin, 573–586 nm; phycocyanin and allophycocyanin, 649–661 nm; chlorophyll a PSII, 674–689 nm; chlorophyll a PSII, 711–727 nm), and the frequency characteristics of the corresponding first-derivative Fourier transforms for each plot (mean values in three specified frequency domains: 43–58 μm-1, 95–110 μm-1, 123–135 μm-1). The detailed description of the extraction of classification parameters is given in Zhangirov et al. [28].
\nLinear discriminant analysis (LDA) is well-known and often applied in biology for various classification problems [15, 17, 29, 30]. Linear discriminant analysis (LDA) is a statistical technique for classifying samples into two or more groups (classes) [31, 32]. It utilizes linear combinations of independent variables to form a basis for a classification scheme. In our case, the independent variables are 63 classification parameters extracted from each set of single-cell self-fluorescent spectra.
\nLDA builds n linear discriminant functions, where n is a number of classes and a row vector with a number of parameters describes each observation. The decision of the sample belonging to the class is based on the selecting of the maximal discriminant function for the sample row vector. Discriminant analysis has two very useful applications. First, it identifies a set of classification parameters that are needed to discriminate between known groups, that is, sets of classification parameters can be identified that are necessary to discriminate between known cyanobacteria strains. Second, the analysis can be used to classify an unknown sample (within a certain probability) into a known group of species or strains. The high classification accuracy of LDA is due to the fact that it works with distribution functions for classification parameters and their statistical characteristics, which allows to build better classification model. However, LDA has strong restrictions on the presence of correlations between classification parameters.
\nIn addition, LDA allows to reduce dimension of the feature space. This so-called linear Fisher discriminant analysis (LFDA) is a data classification method, which classifies the samples by dividing them into groups. The boundaries of these groups are determined by threshold coordinate values. The goal of this method is to find the informative projections by maximizing the function constructed of the projective matrix, the between-class scatter matrix, and the within-class scatter matrix. In this procedure, the first largest component (canonical discriminant function) is the maximal, and the classifications are performed using the three-dimensional space defined by the three largest components. The selection of the best classification parameters is based on the criterion that the dissimilarity between classification parameters of different species/strains should be greater than between those of the same group. Actually, LFDA bases on a solution of eigenvalue problem. The eigenvectors with the first highest eigenvalues are used to construct a lower dimensional space, while the other dimensions are neglected.
\nAlso a stepwise discriminant analysis (SDA) was used in this investigation at the stage of selection of the most valuable classification parameters to determine which parameters discriminate better between the specified groups of observations. Standardized coefficients for each variable in each discriminated function represent the contribution of the respective parameter to the discrimination between groups.
\nThe calculations were performed in MATLAB software using custom-built programs [33].
\nArtificial neural networks (ANNs) are currently being used in a variety of applications with great success [8, 34, 35, 36]. In contrast with conventional programs for data analysis, neural networks follow an adaptive approach. They are flexible and eminently suited for application to complex data structures that are not apt for other data analysis methods like cluster analysis or principal component analysis. Their first main advantage is that they do not require a user-specified problem solving algorithm (as is the case with classic programming), but instead they “learn” from examples, much like human beings. Their second main advantage is that they possess an inherent generalization ability. This means that they can identify and respond to patterns that are similar but not identical to the ones on which they have been trained.
\nANN can be described as a mathematical model of a specific structure, consisting of a number of the single processing elements (called artificial neurons), arranged in interconnected layers. An active neuron multiplies each input vector by its weight, sums the products, and passes the sum through a transfer function to produce the output [37]. The ANN is made up of a group of interconnected artificial neurons, belonging to different layers, while inside one layer neurons are independent. ANN consists of an input, hidden, and output layers. Each neuron transforms input and sends outputs to other neurons to which it is connected.
\nThere are many different types and architectures of neural networks varying fundamentally. In this paper a feed-forward ANN (FFANN) is used for solving considered classification problem [34, 37]. Figure 2 illustrates the model of the ANN used in this work. Due to the simplicity of the classification problem to be solved, a multilayer feed-forward neural network (NN) with one hidden layer was considered. As an activation function, a hyperbolic tangent was used at the hidden layer and Softmax function at the output layer, which allows interpreting the output layer as the distribution of probabilities of belonging to each of the classes. On both layers a bias neuron with a signal equal to unity is added. The size of the input layer (\n
The multilayer feed-forward artificial neural network. The input classification parameters are fed to the input layer of ANN, and signals are propagated through the network via internal neurons to the output layer. In this way input signal pattern and output signal pattern are associated with each other.
Learning in ANNs is accomplished through special training algorithms developed based on learning rules presumed to mimic the learning mechanisms of biological systems. According to supervised learning, the network is trained with a dataset of observations and optimized basing on its ability to predict a set of known outcomes. The deviation of the network solution from the target (true) value is computed, and the calculation of the error is propagated backward from the output layer to adjust the connection weights. Since in our case the activation function at the output layer was determined as Softmax, the loss function was calculated via cross-entropy method. A lot of special training algorithms were developed according to learning rules. In this investigation the method of adaptive moment estimation (Adam) was chosen for further calculations [38].
\nIn the training phase, a sample set of classification parameters and the known solution (the strain number of the corresponding cell) are forced iteratively upon the network. The neuron’s weights (ANN parameters) are adjusted in small steps until the network has learned the training examples. In the experiments described in this study, the training procedure has 500 iterations (epochs). After training, the network is tested. In this test phase, the characteristics of a number of cyanobacterial cells with known identities are fed to the network, and the solutions are compared with these known identities. In this study, after training the network was capable of recognizing about 96% of cyanobacterial cells in the test set. The analysis of generalization quality of ANN is identical to the test procedure; only the identities of the cells are not known beforehand.
\nThe ratio of training sample to the test sampling in this investigation was taken 70:30%. Other parameters of the selected training algorithm were as follows: acceptable error threshold is 0.01, the bandwidth parameter (size of error control window) is 20, the moment parameter is 0.1, and the regularization parameter is 0.001. The selected learning rate was chosen 0.01, and the number of training epochs lays in the range from 300 to 800.
\nThe main criteria for assessing the quality of ANN operation is the value of classification accuracy. There are several approaches to evaluate the accuracy of classification. In the considered case, the classification accuracy is calculated for each class separately, as the ratio of the number of correctly classified class observations to the total number of observations in a given class. Then the average classification accuracy for all classes was obtained. In such case it is possible to build a matrix of errors with size N × N (N—number of classes) and present the results in a bar chart, on which a classification accuracy for each class can be visualized (see Figure 8 in “Ataxonomic differentiation of cyanobacterial strains on the base of single-cell fluorescence spectra”).
\nOn the base of the classification accuracy analysis, it is possible to evaluate the quality of ANN training as well as the quality of internal and external generalization. In our case, the evaluation of the quality of external generalization was obtained on the base a priori knowledge about new species, which was taken from an expert. To validate the correctness of the neural network operation, the results of the NN classification were compared with the results of the LDA.
\nThe ANN architecture presented in this paper, as well as the learning algorithm and its parameters were determined during the study of various configurations. The selected model after training consistently gives a classification accuracy of at least 95%. In this study, ANN was simulated using MATLAB software [33].
\nIn cyanobacteria, the antenna complexes for photosystem II (PS II) and to some extent for photosystem I (PS I) are extrinsic and formed as large multiprotein organelles, which are located on the stromal side of the thylakoid membranes. These supramolecular pigment-protein complexes, so-called phycobilisomes (PBSs), first described by Gantt [39], are the main light-harvesting antennae in cyanobacteria.
\nPhycobilisomes are primarily composed of phycobiliproteins, a colored family of water-soluble proteins. Their chemical and spectroscopic properties are determined by their structure and function that they perform in the photosynthetic process. The three classes of phycobiliproteins are allophycocyanin (APC), phycocyanin (PC), and phycoerythrin (PE). However, in some cyanobacteria phycoerythrin can be replaced by phycoerythrocyanin (PEC), or both pigments can be lacking; phycocyanin and allophycocyanin are constitutively present in all cyanobacteria. Actually, there are very slight species differences between detached phycobiliproteins, even between prokaryotic cyanobacteria and eukaryotic red algae [40].
\nUsually PBSs are assembled from 12 to 18 different types of polypeptides, which may be grouped into three classes: (1) phycobiliproteins, (2) linker polypeptides, and (3) PBS-associated proteins. The amino acid sequences of all components constituting the phycobilisomes of some cyanobacterial strains have been determined, and analysis of these data has revealed phylogenetic relationships [41].
\nThe polypeptide composition of PBS varies widely among strains of cyanobacteria. It should be noted that the degree of PBS compositional variability, which reflects the ability of an organism to adapt to environmental changes, varies from strain to strain. Moreover, for a single strain it sometimes depends upon the environmental conditions such as nutrient availability, temperature, light quality, and light intensity.
\nIt is well-known that total biliprotein content of cyanobacterial cells is inversely related to the quality and quantity of irradiance. A comprehensive review given in Refs. [41, 42, 43, 44, 45, 46] details the various degrees of such chromatic adaptation. However, for cyanobacteria cultured under white light of reasonable intensity and in the medium with habitual nutrient composition, no chromatic adaptation can occur, and PBS structure remains invariable within each strain. Thus the unique spectroscopic properties of different cyanobacterial strains, while analyzing in vivo, may become promising fingerprints for practical and laboratory applications.
\nPhycobilisomes are constructed from two main structural elements: a core substructure and peripheral rods that are arranged in a hemidiscoidal fashion around that core (Figure 3). Each core cylinder is made up of four disc-shaped phycobiliprotein trimers, allophycocyanin (APC), allophycocyanin B (APC-B), and APC core-membrane linker complex (APC-LCM). By the core-membrane linkers, PBSs are attached on thylakoids and structurally coupled with PSII. The peripheral cylindrical rods (six or eight) radiate from the lateral surfaces of the core substructure and are usually not in contact with the thylakoid membrane. The rods are made up of hexamers, disc-shaped phycobiliproteins (PE, PEC, and PC), and corresponding rod linker polypeptides [41, 42, 43, 44]. Most linker polypeptides are colorless proteins, but some also contain phycobilin chromophores, endowing them with the ability to harvest light as well as aid in the assembly of the phycobilisomes [46]. For more details about phycobilisome structure, see [18, 39, 47].
\nSchematic drawing of phycobilisome and photosynthetic energy transfer to reaction center.
The phycobilisome is attached to the membrane by multiple weak charge-charge interactions, either with proteins or with lipid head-groups. Binding is rather unstable. The core-membrane linker polypeptide provides a flexible surface, allowing interaction with a range of structurally distinct membrane complexes, including photosystem II (PSII) and photosystem I (PSI) (see Figure 4). The stability of each interaction may be modulated by covalent modification and/or the presence of accessory subunits.
\nModel for the interaction between phycobilisomes and membrane components. (a) Phycobilisomes are shown as semicircles, with the core as a darker rectangle [
Recently, it was established that phycobilisomes diffuse rapidly on the surface of the thylakoid membrane, while PS II reaction centers are normally almost immobile. Fluorescence recovery after photobleaching (FRAP) has been used to measure the mobility of phycobilisomes in the intact cyanobacterial cells [48, 49], and it was clearly demonstrated that a significant proportion of phycobilisome-absorbed energy is delivered to PS I as well as to PS II [45, 49, 50].
\nThe high mobility of phycobilisomes along the thylakoid membrane gives the opportunity of the occasional direct interaction of phycobilisome rods or core with PS I (Figure 4). Two ways that energy could be transferred from phycobilisomes to photosystem I are shown in Figure 4; “Spillover” from photosystem II with an attached phycobilisome (supposed by Su et al. [51]) (Figure 4a, left photosynthetic complex) and direct association of the phycobilisome core with photosystem I (Figure 4a, right photosynthetic complex).
\nAnother possible variant of the interaction between phycobilisome and reaction centers of two photosystems was proposed by Gantt in the Chapter 6.3 of the book [40]. The author assumed that the special close arrangement of both photosystems around the base of the phycobilisome provides the partial transfer of the absorbed energy to PSII and PSI simultaneously (Figure 4b).
\nThe intrinsic fluorescence of photosynthetic organisms originates from excited states that were trapped by light-harvesting system and lost before photochemistry took place. Photoexcitation energy absorbed at the outer surface of phycobilisomes is transported sequentially through several rod chromoproteins to an inner core and then to core-membrane linker (the terminal pigment) that acts as the final energy transmitters from the phycobilisome to Chl a heterodimers of two photosystems (PSII and PSI), incorporated in the thylakoid membrane. This excitation transfer is recognized as due to the Förster dipole-dipole interaction with an extremely high efficiency, near unity.
\nThe more distal parts of the antenna system, a peripheral antenna complex (phycobilisome), maximally absorb photons at shorter wavelengths (higher energies) than do the pigments in the antenna complexes that are proximal to the reaction center. Subsequent energy transfer processes are from these high-energy pigments physically distant from the reaction center to low-energy pigments that are physically closer to the reaction center (Figure 5). With each transfer, a small amount of energy is lost as heat, and the excitation is moved closer to the reaction center, where the energy is stored by photochemistry. Note that the probability of excitation energy escape from the trap in the form of fluorescence at all transfer steps is non-zero and depends on the intensity and wavelength of the excitation light.
\nSchematic illustration of the energy transfer in light-harvesting system of cyanobacteria (a). Panel (b) represents the example of normalized in vivo single-cell fluorescence emission spectra at excitation wavelength 488 nm. Dashed lines and letters over them indicate emission wavelengths of PE, PC, APC, and Chl a of PSII and PSI, correspondingly. Fine lines represent fluorescent spectra of corresponding detached pigment-protein complexes.
During the energy transfer process, the occasional quenching of the absorbed light by fluorescence can occur, and this becomes the essential property for fluorescent spectroscopy. It usually represents a small fraction of the excited states and diminishes in a functioning photosynthetic complex. Nevertheless, the fluorescence is an extremely informative quantity, because it reports on the energy transfer and trapping. Both steady-state and time-resolved fluorescence measurements are widely used methods for probing the organization and functional state of photosynthetic systems.
\nThe fluorescence of intact living cyanobacterial cells is originated from the inefficiency of the energy transfer between all components of the energy transfer chain including the final step, the delivery to PSII or PSI (Figure 5a). Due to the occasional quenching by fluorescence each transfer step result in peak or shoulder on the corresponding spectrum (Figure 5b). This is due to the fact that when phycobilisomes are bound to the thylakoid membrane, most of the energy from phycobilisome is channeled to chlorophylls in the thylakoid membrane and thus did not shade the fluorescence of the previous steps in energy transfer chain. In the course of the energy transfer from the initially photoexcited phycobiliprotein to the reaction center of photosystems PSI and PSII, fluorescence is emitted from almost every type of pigment and can be used as a probe to examine the mechanism of energy transfer within the light-harvesting system [43, 44, 52].
\nA convenient way to monitor this energy transfer process is to irradiate a sample with light that is selectively absorbed by one set of pigments and then monitor fluorescence that originates from a different set of pigments. Obviously, if the energy transfer is taken place between pigments, the light absorbed by one set of pigments is emitted by another set differently, depending on the excitation wavelength. This type of fluorescence excitation experiment can be used to measure quantitatively the efficiency of energy transfer from one set of pigments to another [43]. Moreover, different species of cyanobacteria contain different accessory pigment proteins and specific linker proteins between them; therefore a set of fluorescence emission spectra excited by different wavelengths have its own unique shape for the cells of one strain and are quite distinguishable from other species and strains. Such sets of fluorescence emission spectra can be used for automatic differentiation of cyanobacterial species.
\nFigure 6 shows several characteristic sets of single-cell fluorescence spectra corresponding to Microcystis CALU 398, Merismopedia CALU 666, Leptolyngbya CALU 1715, and Phormidium CALU 624, obtained by confocal laser scanning microscope (CLSM) Leica TCS-SP5, which are placed near each set. Each spectrum in the set was obtained using different laser lines for excitation: 405, 458, 476, 488, 496, 514, 543, and 633 nm. Corresponding excitation wavelengths are given over each spectrum. All spectra are normalized to the maximum intensity and shifted along x-axis for convenience of observation. It can be easily noticed that laser line 458 nm excites mostly in vivo fluorescence of Chl a in both photosystems PSII and PSI around 682 and 715 nm, correspondingly, and the emission spectrum by cyanobacterial cells shows no appreciable emission of PC or APC. In cyanobacteria, the 458 nm excitation is preferentially absorbed by PSI that contains more Chl a than by PSII and is stoichiometrically more abundant than PSII. However, because reaction center of PSI turns over faster than the PSII, it has lower fluorescence intensity than the PSII antenna. This is indicated by PSI emission band at 715 nm which is much weaker than the PSII emission band at 682 nm. The excitation by intermediate (blue and green) wavelengths (405, 488, and 496 nm) reveals fluorescent maxima of all photosynthetic pigments, as the light in this range is absorbed by all pigment-protein complexes almost in equal portions and fluorescence emits by all steps of energy transfer chain (Figure 5). The direct excitation of cells in the PE absorption region at 514 and 543 nm results in emission spectrum with two main peaks at 580 and 656 nm, which are due to PE, PC, and APC emission, and for species that lack PE, the emission accumulates mostly near 656 nm. Two chlorophyll fluorescence components can be resolved for some species in a number of spectra. The spectra of the 633 nm excitation directly give a prominent emission band at 656 nm that originates from C-PC, omitting band at 580 nm, which cannot be excited by 633 nm, even for species that have PE (see Figure 6). Other small emission bands, corresponding to fine pigment structure of antenna complex, are not resolved at the room temperature.
\nFour characteristic sets of single-cell fluorescence spectra. The excitation wavelengths (405, 458, 476, 488, 496, 514, 543, and 633 nm) are given over the curves. All spectra are normalized to the maximum intensity and shifted along x-axis for convenience of observation. The dashed lines indicate fluorescence maxima of the individual pigments (PE, 580 nm; PC, 656 nm; Chl a, 682 and 715 nm).
These in vivo fluorescence emission spectra reflect the structure of light-harvesting complex of corresponding species and correct or incorrect functioning of its energy transfer chain. Four characteristic wavelengths, corresponding to the fluorescence maximum or shoulder, can be easily distinguished: (1) peak near 580 nm corresponds to the fluorescence of phycoerythrin, (2) peak near 656–560 nm corresponds to the fluorescence of phycocyanin and allophycocyanin in common (they are undistinguishable at room temperature), (3) peak near 682 nm corresponds to the fluorescence of chlorophyll a in photosystem II, and (4) peak or shoulder near 720 nm represents the fluorescence from photosystem I [10, 53].
\nComparative analysis of the series of fluorescence spectra for different cyanobacterial species and strains reveals visible variations in their shape. If the fluorescence spectra were taken from live cells in normal physiological state, which are cultured in the same growth environmental conditions, then the interspecies variations in pigment/Chl a ratios are more pronounced than variations within the individual species. And species/strain differentiation could be carried out on the basis of conventional multivariate analysis.
\nFluorescence spectra have been used to classify phytoplankton populations since approximately the early 1970s [54, 55]. However, because of the generally low device precision and poor availabilities, the rate of species discrimination was relatively low. Recently new attempts to conduct the discrimination among microalga on the base of absorption or fluorescence spectra were reported [7, 10, 13, 15]. But again in published experiments only big algal groups with a considerable differences in pigment composition can be successfully separated (e.g., cryptophytes, chlorophytes, cyanobacteria, etc.). Moreover, all the authors pointed out that the discrimination among cyanobacterial species is quite complex and ambiguous. Actually, the correct discrimination of cyanobacterial species on the base of fluorescence signature is usually hampered by alterations in the pigment composition within one strain, which depends on the environmental conditions and physiological state of the culture. These difficulties can be overcome by using single-cell fluorescence spectra instead of bulk ones and by recording 7–8 spectra with different excitation wavelengths for each cell instead of one or two as usually is done.
\nIn the presented investigation, 307 sets of 8 single-cell fluorescent spectra for 23 cyanobacterial strains, belonging to 15 genera, were analyzed. An optimal set of classification parameters was considered that is sufficient for determining the generic membership of cyanobacterial cells by means of mathematical statistics. The results of this study show that LDA and ANN are able to recognize cyanobacteria up to species/strains according to the data recorded by means of CLSM. This implies that the classifier (LDA or ANN) is capable of defining a unique niche in a multiparameter space for each of 23 cyanobacterial strains, used in this investigation.
\nThe results of LDA, evaluated over 63 parameters extracted from 307 single-cell fluorescence spectra, are presented in Figure 7 as 3D-plots in the space of canonical discriminating functions. It is clear that the discrimination between species is sufficiently good. Moreover, the closely related species (e.g.,
The results of linear discriminant analysis. (a) and (b) Observations in 3D space of first three maximal canonical discriminant functions (root 1,2,3). Solid curves bounded the regions, occupied by seven new species. In the legend new species are indicated with red. (c) Classification diagram for 23 cyanobacterial strains from CALU collection. Red dots indicate false results.
In the legend all used cyanobacterial strains are named and enumerated according to CALU collection. Solid curves bounded the regions, occupied by seven strains (
In the considered classification problem, the quality of the ANN operation should be determined not only by the absolute value of the classification accuracy but also by the ability of the designed ANN to recognize and properly classify unknown species that did not participate in the training process. Thus, the performance of ANN was tested first with the aim only to discriminate between 16 known cyanobacterial species (Figure 8a). Another seven strains were identified as test ones, to verify the correctness of ANN in recognizing new strains (so-called generalization quality). Analysis of a test set with data from the same monocultures confirmed that the parameters extracted from the fluorescent spectrum sets contained enough information to correctly identify cyanobacterial cells at the species/strain level. The trained neural network presented here showed not the highest rate of correct classification—only about 95.7%—but it shows the best recognition quality for new strains. The results of the ANN recognition are presented in Figure 8b.
\nThe results of ANN classification. (a) The results of recognition of 16 known cyanobacterial strains. (b) The results of recognition of seven unknown strains. Numbers over each bar indicate maximal class probability for each strain. (c) General classification results. Red dots indicate false results. Strains are numbered according to CALU collection names.
Bar charts in Figure 8a represent the results of the classification of 268 experimental measurements by 16 classes. Each bar represents the classification results as the probability distributions. Each color in the bar corresponds to 1 of 16 target classes (known cyanobacterial strains). The percentage rate of colors in the bar shows the probability distribution of belonging to the target classes. Maximal eigenclass probability is indicated above each bar.
\nIn contrast to standard classifiers, a classifier built on the base of ANN has a so-called generalization ability. It means that ANN is able to recognize new cyanobacterial strains that were previously unknown for it and suggest possible variants of their generic affiliation to known classes. In Figure 8c, the ANN classification results for 16 target classes and 7 strains that were not presented in the training set are shown. The aim of ANN classifier was to determine which of the 16 known classes and 7 unknown strains could be attributed. The results of ANN classification correlate well with the results predicted by LDA (Figure 7). The closely related strains in this case were 1763–666, 601–398, 756–1409, 1315–1718, 550–1416, 824–1817, and 1336–398 (in the pairs, the first strain is unknown for ANN, and the second is the one of the nearest target classes). The strains of 1336
To validate the correctness of the neural network operation, the results of the ANN classification were compared with the results of the LDA. The neural network-based classification agrees well with the expected results and with the results of LDA. The identification performance of the network for cyanobacterial strains from the same species is slightly less than for the cells from different species, but anyway they can also be distinguished perfectly well.
\nThe automatization of the cyanobacterial species differentiation is a key problem in both industrial biomass production and environmental monitoring. Unfortunately, all presently utilized methods cannot be implemented in online monitoring procedures due to various reasons. In this work, an example of the use of LDA and ANN technologies for online differentiation of cyanobacterial strains according to their in vivo single-cell fluorescence spectra is presented. The novel discrimination technique demonstrated here includes a strict procedure for recording and processing single-cell fluorescence emission spectra, which eliminates most of usual data processing difficulties and, as a result, has a quite high classification accuracy. And the initial information is obtained via fluorescent spectroscopy; the experimental data can be processed automatically. Moreover, due to the use of CLSM microscopic spectroscopy instead of conventional fluorimetry, the initial data have less variations and can be accurately sorted. Any objectionable and unpredictable impact is eliminated at the first step of obtaining fluorescence spectra. Since noninvasive and nondistructive method is used, the information about vital cell operation (e.g., light harvesting) can be additionally taken into account, to obtain the desirable precision of discrimination.
\nThe universality of the considered technique makes it possible to use it for investigation of any phytoplankton species irrespective of their habitat or cultivation. Utilizing data from several fluorescence spectra, instead of one, results in more fingerprint information which leads to the taxonomic differentiation on a finer scale. Differentiation procedure, presented here, was carried out by means of statistical analysis on the base of mathematical characteristics of intrinsic fluorescence spectra of living single cells; therefore it is free from usual subjectivity, which can occur while using methods of direct optical microscopy. Moreover, formalization of data processing gives a wide opportunity for automating of the classification procedure of cyanobacterial strains in field samples, while online monitoring of water bodies is conducted.
\nUndoubtedly, the data set should be expanded to include more species and phytoplankton classes/divisions, grown under different nutrient and light conditions. However, this work already demonstrates the potential of the discrimination of phytoplankton classes by means of fluorescence microscopic spectroscopy. Combining the knowledge of phytoplankton structure along with taxon-specific measurements of photosynthetic activity and biochemical cell composition can lead to new models which increase the reliability of online monitoring.
\nWith the growing concerns on global warming and energy prices, the demand for environmentally friendly vehicles with better fuel efficiency is increasing [1, 2, 3]. These issues can be addressed by reducing car weight, lowering travel resistance, advancing drive-train efficiency, developing new sources of power, and so on. Vehicle weight reduction using advanced lightweight structural materials, such as Mg, Al, and Ti, is considered as one of the most promising strategies to address these issues [4, 5, 6, 7, 8, 9, 10]. Generally, for every 10% weight reduction, the specific fuel consumption could reduce by 3–7%, while maintaining the same functionality [11, 12]. Thus, the use of lightweight alloys as structural materials is considered as the factor for development of aerospace and automotive manufacturing sectors in the future. To achieve lightweight, safety, and low cost, the multi-material structure using steels, Al, Mg alloys is considered to be efficient [9]. Therefore, effective dissimilar joining process of such light metals and steels is essential.
As the lightest structural material, Mg alloys receive great attention due to their high specific strength, sound damping capabilities, hot formability, good castability, recyclability among others [13, 14, 15, 16, 17, 18]. Potential applications of Mg alloys in an automobile include seat components, bracket carrier, roof, bonnets, cylinder head, wheels, etc. [8, 19]. Steel is currently the automaker’s material of choice, due to its inherent properties, including high strength and toughness, good ductility, and low cost [8, 20, 21, 22]. Recently, it has been demonstrated through the next-generation vehicle project that stainless steels are promising candidates for vehicle construction, and they can be used replaced carbon steels, especially in crash-relevant components such as door pillars.
Therefore, for practical applications in automotive industries, Mg alloys will have to be joined with existing steel parts. Recently, many automotive components have been produced using a combination of Mg alloys and steels, but the major issues arise from the joining techniques and corrosion of the joined parts [23, 24, 25, 26]. Thus, attaining reliable Mg/steel hybrid joints is paramount for facilitating lightweight industrial fabrication and expanding the industrial applications of Mg alloys in automotive industries [21, 27, 28, 29].
Joining Mg alloys directly to steel is extremely difficult because of the huge differences in their physical and metallurgical properties, the lattice mismatch between Fe and Mg is very large and there is almost zero solubility between Mg and Fe [27, 29, 30, 31, 32, 33, 34, 35]. Hence, an appropriate technique that overcomes the aforementioned problems is very much desired.
Generally, a successful joint between Mg and steel can be achieved by inserting an intermediate material at the interface or diffusion of alloying elements from the BM. At present, several authors have focused on joining magnesium alloys to different grades of steel, using various welding technology, such as friction stir welding (FSW), ultrasonic spot welding (USW), diffusion and eutectic bonding, resistance spot welding, laser welding brazing, laser-TIG hybrid welding and gas metal arc weld-brazing. In these studies, various interlayer elements and alloys such as Zn, Ni, Cu, Cu-Zn, Sn, Al, and Ag have been explored. In contrast to direct joining of magnesium to steel, which is mainly a mechanical bonding, with insertion of the interlayer elements, formation of intermetallic phases or solid solutions between Mg and the interlayer and also the interlayer and Fe indicated that metallurgical bonding is achieved. However, the joint performance and the interfacial bonding achieved depend significantly on the IMC phase formed [31, 36, 37, 38]. To control the morphology and existence state of the intermediate phase, the selection of suitable interlayer material and joining techniques are essential. Generally, choosing the suitable interlayer for joining Mg alloys to steel largely depends on the interlayer composition that gives excellent wetting and bonding without generating thick layers of hard and brittle IMCs at the joint interface [31, 35, 39, 40, 41]. Moreover, when choosing the joining process that will be used, minimization of the thickness of any brittle intermetallic compounds along the interfaces of the magnesium alloy-interlayer-steel joint and minimization of intermixing between the Mg and Fe in the molten-state are the main factors that must be considered [8, 21, 42, 43].
Currently, a great deal of research has been conducted on the interface characteristics and mechanical performance of Mg alloys to steel joints, particularly under static loading. Under optimized processing conditions, excellent static strength has been achieved, even surpassing that of Mg alloy base metal with insertion of Ag, Cu, and Ni intermediate elements [38, 44]. However, few experiments have been carried out on the corrosion behavior of the jointed parts and the joints performance under dynamic loading [45, 46, 47, 48].
With the continuously increasing usage of Mg alloys in industries and the large number of potential applications of Mg/steel hybrid structures, two of the specific areas of concerns for broader utilization of magnesium alloys are reliable joining techniques and corrosion behavior of the joined parts. To better understand and address these challenges, there is a need to comprehensively review the research conducted so far and provide the most efficient strategies to address the challenges. This paper presents a review on Mg alloys/steel joining techniques, with focus on the techniques used to control the morphology and existence state of intermetallic compound (IMC) and improving mechanical properties. The general motives behind this review are to obtain a better understanding on the weldability issues associated with joining magnesium alloys to steel. It would also establish global, state-of-the-art welding techniques of Mg alloys to steel.
Some of the inherent properties of Mg include high thermal conductivities and coefficients of thermal expansion, large solidification temperature ranges, strong tendency to oxidize, low viscosity and surface tensions, high solidification shrinkage, low melting and boiling temperatures, a tendency to form low melting point constituents and high solubility for hydrogen in the liquid state [8, 30, 49]. It is obvious that the properties of Mg differ significantly from those of Fe. For instance, the melting points of magnesium and iron are 649°C and 1536°C, respectively. This wide discrepancy in melting points makes it very difficult to melt the base materials at the same time as might be required in fusion-welding process [21].
In addition, the crystal structure of Iron at room temperature is body-centered cubic (BCC), whereas that of magnesium is close-packed hexagonal (HCP). Crystallographic analysis has shown that the lattice mismatch of Fe and Mg is very large [34, 50, 51]. Although the welding process itself is a non-equilibrium process, phase diagram has always been an effective tool to predict the reactions formed during welding process and serves as a reference to examine the feasibility of achieving a metallurgical bonding between the metals. According to the Mg/Fe binary phase diagram, the maximum solid solubility of iron in magnesium is 0.00043 wt.% while that of magnesium in iron is nil, and the Mg concentration at the eutectic point is estimated to be less than 0.008 at.% [34, 52, 53, 54]. Therefore, magnesium and steel are immiscible (neither the formation IMC nor atomic diffusion occurs between them after solidification), thereby presenting difficulty in joining them together.
In recent years, numerous techniques have been applied to bond Mg alloys to various grades of steels. These techniques can be broadly classified into solid-state joining (friction stir welding, ultrasonic spot welding, diffusion-welding processes) and fusion welding (resistance spot welding, laser welding brazing, laser hybrid welding, and arc welding). Review of the literature reveals that metallurgical bonding along the Mg/steel interface can be achieved with addition of suitable interlayers (which possess a substantial solid solubility in both Mg and Fe) or inter-diffusion of the alloying element from the BM. Therefore, the joint quality is significantly influenced by the interlayer characteristics (forms, thickness, and compositions). Among the joining techniques, a variety of thin interlayers such as Zn, Cu, Al, Ni, Sn, Cu-Zn, and Ag has been reported to improve the interfacial reaction between magnesium alloys and steel. In this section, the potential of several methods for joining magnesium and steel will be discussed.
Solid-state joining technology has been applied to bond Mg alloys to steel and get high-quality joints than fusion-welding processes because of the added advantage of minimal oxidation because of the solid-state nature of the process. Solid-state joining processes, such as friction stir welding (FSW), ultrasonic spot welding (USW), and diffusion and eutectic bonding, have been used to join Mg alloys to steel. Generally, for solid-state bonded Mg alloys-interlayer-steel joints, the joint performance is influenced by the intimate contact between the dissimilar materials, and the microstructure, particularly the formation of IMCs [55, 56].
FSW is a solid-state welding technique invented by Thomas et al. [57]. The combined action of pressure and stirring during the FSW led to mutual diffusion of the alloying elements from the BM, which promoted metallurgical bonding at the interface and improved the joint performance. Some of the unique advantages of FSW include low distortion and residual stresses due to low heat input and absence of melting, filler metal is not required and the heat efficiency is very high relative to traditional fusion-welding processes [58, 59]. Joining Mg alloys to steel by FSW has been extensively studied.
Watanabe et al. [60, 61] investigated the weldability of AZ31 Mg alloy to uncoated SS400 steel joints by FSW. Under optimum joining condition (0.1 mm tool pin offset toward steel plate, 1250 rpm pin rotation speed, and welding speed of 100 mm/min), a joint with maximum strength of about 70% of the Mg BM was obtained. The low joint strength was associated with insufficient plasticization and presence of steel fragments in the Mg matrix. Considerable number of authors observed that the presence of zinc coating on the steel surface played an important role in the bond formation between the Mg alloys and steel [21, 27, 56, 62, 63]. For instance, Schneider et al. [63] and Jana et al. [27] joined AZ31B alloy to Zn-coated steel by FSW. It was found that the Zn coating enhanced the bondability of the Mg alloy/steel. The authors noted that the strength of the FSW welded AZ31B/galvanized steel was significantly superior to that of AZ31B/uncoated steel. The presence of Zn coating promoted the formation of liquid low melting Mg-Zn eutectic products at the interface. The liquid products, broken oxides, and other contaminants were forced out of the joining interface by the high pressure produced by the tool, exposing the fresh interfaces. As a result, mutual diffusion between magnesium alloy and steel was achieved.
Friction stir spot welding (FSSW) a variant of FSW, has also been used to join Mg alloys to steel. Liyanage et al. [29] joined AM60 to DP600 dual-phase steel by FSSW, and the welds showed no evidence of intermetallic formation. They reported that a Zn layer on DP600 steel resulted in melted eutectic material (αMg + MgZn) and cracking in the joints. The feasibility of joining 3 mm thick AZ31 Mg alloy to 1 mm 302 stainless steel by FSW was also reported [64]. The results showed that the combined action of pressure and stirring led to mutual diffusion of the alloying elements from the BM at the interface. Under optimum condition, the joints shear strength reached a maximum of 96.3 MPa. However, void and microtype defects at the interface were observed.
Joo et al. [65] reported that an acceptable AZ31B/SS400 steel joints with higher strength, sufficient material flow and tool wear reduction were obtained using hybrid gas tungsten arc welding (GTA) and friction stir welding. The tensile strength of the hybrid friction stir welds increased to 237 MPa about 91% of the Mg BM, compared to 226 MPa for conventional friction stir welds.
The prior research proved that refill friction stir spot welding (RFSSW) has many advantages for joining of Mg alloys/steel dissimilar welds [48, 66, 67]. Zhang et al. [67] joined 3 mm AZ31B to 1 mm galvanized steel joints by friction stir keyholeless spot welding (FSKSW) and reported that the stacking sequence of the workpieces played a significant role in determining the mechanical properties. Under optimum joining condition, the maximum joint strength of 8.7 kN was achieved. Furthermore, 1.53 mm ZEK100 Mg alloy was welded to 1 mm Zn-coated DP600 steel with 10 μm Zn coating by RFSSW [66]. The authors observed that the sleeve did not plunge into the bottom steel. A thin interfacial layer with thickness of <100 nm identified as FeAl2 by TEM was observed at the interface, which accommodates bonding between the immiscible Mg and Fe and appeared to have originated from Zn-based galvanized coating on steel. Under optimum process parameters (tool speed of 1800 rpm, welding time of 3.0 s, and penetration into the upper ZEK100 of 1.5 mm), the joints shear strength of 4.7 kN was achieved.
However, numerous studies focused on characterizing the Mg alloys to steel joints under dynamic loading condition [20, 48, 68]. For example, the fatigue behavior of FSW-produced by AZ31 and two types of Zn-coated steels (HSLA steel or mild steel (MS)) lap joints has been investigated [20]. It was found that the performance of friction stir welded joints under fatigue load is limited due to ‘hook’-like features formed along the magnesium/steel interface, which act as stress raisers. Uematsu et al. [68] investigated the static and dynamic behavior of dissimilar lap joining of 2 mm AZ31 Mg alloy to 2 mm cold-rolled low carbon steel by FSSW. The results showed that fatigue crack grew through the interface, regardless of load levels. The authors noted that the effective nugget size could be the controlling factor of the fatigue strengths of dissimilar welds, and it is essential to increase the effective nugget size to improve the fatigue performance of dissimilar welds. Shen et al. [48] compared the static and dynamic behavior of 1.5 mm ZEK100 Mg alloy/0.9 mm galvanized DP600 steel dissimilar spot welds to Mg/Mg similar welds produced by RFSSW. It was observed that the static and dynamic behaviors of the magnesium/steel welds were superior to that of Mg/Mg similar spot welds. The higher joint performance observed for Mg to Zn-coated steel was associated with an increase in effective bonded area, compared to Mg/Mg spot welds due to the presence of displayed Zn-coated layer. Furthermore, the analysis of the dynamic behavior of the Mg alloys to galvanized steel joints revealed that the Zn brazing quality directly influences the fatigue life. Therefore, optimization of the welding parameters to facilitate this brazing to magnesium is necessary [48].
Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by FSW and FSSW are shown in Table 1. In FSW welding, prior studies placed Mg alloys as the top sheet while the tool plunge depth was carefully controlled to avoid contacting the steel tool to the bottom ferrous substrate. However, this arrangement resulted in a low joint strength [64]. Interestingly, with recent RFSSW, the sleeve did not lunge into the bottom steel, which significantly increased the effective bond area and improved the joints performance [66, 67, 68, 69, 70, 71]. During FSW and FSSW, the processing heads impose size and shape limitation, the process is best used with long and straight welds, and keeping the interlayer at the interface between the steel and Mg alloy is very difficult due to the stirring action of the pin and material flow with high plasticity along the interface. Thus, limited the wide range of application of these techniques in industries [48, 72]. The comparison of the joint properties reveals that good static weld strength can be obtained between magnesium alloys to galvanized steel by a diffusion and braze-bonding mechanism [48, 66]. Most studies focused on using Zn interlayer, and there is a need to experiment with other interlayers. Furthermore, no study focused on the corrosion behavior of the joined parts.
Techniques | Materials | Joint design | Transition material | Maximum tensile shear strength (MPA) | Failure mode | References |
---|---|---|---|---|---|---|
FSW | 2 mm AZ31B-O/2 mm SS400 mild steel | Butt (Offset 0.1 mm toward steel) | No interlayer | 178.5 MPa | Interfacial | [60, 61] |
FSW | 1.6 mm AZ31/0.8 mm Zn-coated steel | Lap (Mg on top) | Zn coating | 3.7kN | Interfacial | [21, 62] |
1.6 mm AZ31/0.8 mm Brushed finished steel | Uncoated | 2.9kN | Stir zone | |||
FSW | 2.33 mm AZ31B/1.5 mm HDG steel | Lap (Mg on top) | Hot dip Zn coating | 6.3 ± 1.0 kN | Interfacial | [27] |
2.33 mm AZ31/0.8 mm HSLA electrically galvanized steel | Electro-galvanized Zn coating | 5.1 ± 1.5 kN | ||||
FSW | 2 mm AZ31B/2 mm DX54D | Lap (Mg on top) | Uncoated | 98 MPa | Stir zone | [63] |
Zn coating (15 μm thick) | 158 MPa | |||||
FSSW | 1.2 mm AM60/1.8 mm DP600 | Lap (Mg on top) | Zn coating | 2.4 ± 0.5kN | AM60 BM | [29] |
FSSW | 2 mm AZ31 Mg alloy/2 mm cold-rolled low carbon steel | Lap (Mg on top) | No interlayer | 32 MPa | Interfacial | [68] |
FSW | 3 mm AZ31/1 mm SUS302 | Lap (Mg on top) | No interlayer | 96.3 MPa | Stir zone | [64] |
GTA-FSW | 3 mm AZ31B/3 mm SS400 | Butt | No interlayer | 237 MPa | Interfacial | [65] |
FSW | 226 MPa | |||||
FSKSW | 3 mm AZ31B/1 mm Q235 | Lap (Steel on top) | Zn coating | 8.7kN | Not reported | [67] |
RFSSW | 1.55 mm ZEK100/1 mm DP600 | Lap | Zn coating | 4.7kN | Interfacial | [66] |
RFSSW | 1.5 mm ZEK100/0.90 mm DP600 | Lap | Pure Zn coating (10 μm thick) | 3.6kN | Interfacial | [48] |
Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by FSW and FSSW.
USW is also a solid-state welding technique that generates coalescence through a concurrent application of localized high-frequency vibratory energy and slight clamping force [73]. At present, the research conducted on joining magnesium alloys to steel using USW is limited.
Santella [74, 75] joined 1.6 mm thick AZ31B-H24 to 0.8 mm HDG mild steel by USW. The authors noted that the presence of Zn improved the bonding mechanism, but the Mg-Zn phases were completely squeezed from the spot weld and only thin Al5Fe5 phase was formed at the interface. Under optimum welding parameters, a joint with maximum lap shear strength of 4.2 kN was achieved. In another related study, Patel et al. [73] also noted that Mg and Zn combined to form Mg-Zn IMCs, while Fe and Zn combined to form a solid solution to create the weld joint. The results of these studies could suggest that no melting on the steel side occurs during USW. In comparison, Patel et al. [76] observed that the shear strength of the magnesium alloy/bare steel with the addition of Sn interlayer was higher than that of magnesium alloy/bare steel and magnesium alloy/galvanized steel joints, due to the solid solutions of Sn formed with magnesium and iron, as well as Sn and Mg2Sn eutectic.
On the other hand, it was demonstrated that corrosion could impair the mechanical performance of Mg alloy/steel USW joints [46]. However, the details of corrosion mechanisms required further study.
Table 2 compares maximum tensile shear strength of the Mg alloys/steel joints produced by USW. The USW techniques involved the use of an interlayer to achieve interfacial reactions. The highest joint strength (about 88% of Mg/Mg ultrasonic spot weld) was obtained using Sn interlayer due to the solid solutions of Sn formed with magnesium and iron, as well as Sn and Mg2Sn eutectic structure. Therefore, using Sn interlayer resulted in better joining mechanism and mechanical performance and is thus more suitable [76]. Thus, choosing of suitable interlayer is essential for successful joining. The possibility of using different transition materials should be explored. Furthermore, studies on dynamic and corrosion behavior of the USW joint parts should be given attention.
Techniques | Materials | Joint design | Transition material | Maximum tensile shear strength | Failure mode | References |
---|---|---|---|---|---|---|
USW | 1.6 mm AZ31B-H24/0.8 mm HDG mild steel | Lap | Zn coating (9 μm thick) | 4.2kN | Through the AZ31 BM | [74, 75, 77] |
USW | 2 mm AZ31B-H24/0.8 mm HSLA | Lap | Zn coating (10 μm thick) | 47 MPa | Interfacial (cohesive failure) | [76] |
Uncoated | Sample failed during specimen mounting | — | ||||
Uncoated with Sn interlayer (50 μm thick) | 71 MPa | Partial nugget pull-out |
Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by USW.
Diffusion methods have been used to successfully join both similar and dissimilar materials combinations, including Mg alloys/steel [37, 78]. During the diffusion process, formation of uniform and thin IMC along the Mg alloy/steel interface is the key for successful bonding. The number of research suggested that interfacial bonding could be achieved by either addition of interlayer or some alloying elements in the Mg BM. The benefits of using Cu [78, 79, 80], Ni [37, 80], and Ag [44, 81] interlayers during diffusion bonding of Mg alloy/steel have been demonstrated.
Tachibana et al. [82] studied the influence of the Zn insert on the bondability of Mg/steel lap joints. It was found that the AZ31 and cold-rolled steel plate (SPCC) could not bond due to the oxide films formed on the AZ31 that prevented the bonding, while AZ31/Zn-coated steel (GI) was bonded successfully. The presence of zinc coating enhanced the bondability of Mg alloy/steel and removed the oxide films on AZ31. The weldability of 5 mm AZ31 Mg alloy to 1 mm thick 316 L steel joints using a diffusion brazing process with addition of Cu and Ni interlayers was also reported [37, 79, 80, 83]. Solid-state diffusion of the interlayer into the magnesium alloy, eutectic formation, and the formation of ternary IMCs was observed at the joint interface. The interfacial reaction was intense in the liquid state, inducing the excessive formation of brittle and thick IMC layers, which was detrimental to the joint strength. The maximum joint strength of 57 MPa (69% of that of AZ31 BM) with Cu interlayer was obtained compared to 32 MPa for Ni added joints. The high interfacial bond obtained for Cu-added joint was associated with confined intermetallics at the bonded interface compared to dispersed intermetallics for Ni added joint [80].
Some authors focused on bonding magnesium alloys to steel using reactive transient liquid phase bonding (rTLP) to improve the joints strength between dissimilar metals. During the rTPL process, eutectic melting and subsequent isothermal solidification occurred between the interlayer and the magnesium, while the formation of a thin continuous layer between the melt and the steels resulted in interfacial bonding of the steel with the magnesium substrate [44, 81]. The bondability of AZ31 to low carbon steel with the addition of Ag interlayer using rTPL process showed that isothermal solidification of the eutectic melt was formed at the Mg alloy side through the diffusion of Ag into the magnesium BM. Thin and uniform Fe2Al5 layer was observed at the steel side, which significantly improved the joint strength to 201 MPa [44, 81]. In contrast, coarse, non-uniform IMC intermittently formed at the interface without Ag interlayer, which deteriorates the joint performance.
Table 3 gives a comparison of the Mg alloys/steel joints maximum tensile shear strength produced by diffusion and eutectic bonding. The literature reveals that the interlayer material has significant influence on the bondability of the Mg alloys to steel by diffusion and eutectic bonding process. The use of Cu, Ni, and Ag as intermediate elements has been studied. The major challenge associated with this technique is that the formation of ternary IMCs which could not be controlled and have detrimental effects on the joint performance [80]. A comparison of the joints strength shows that excellent static strength of Mg/steel joints has been achieved using rTPL techniques, even surpassing that of AZ31B Mg alloy BM with addition of Ag interlayer [44]. The rTPL unique qualities of short process time, coupled with forming a thin and continuous intermediate layer through the formation of a transient liquid interlayer, were responsible for the high joint performance obtained. The possibility of improving the joint performance using more interlayers should be explored.
Techniques | Materials | Joint design | Transition material | Maximum tensile shear strength (MPa) | Failure mode | References |
---|---|---|---|---|---|---|
Diffusion bonding | AZ31/SPCC | Lap | Uncoated | — | No reported | [82] |
AZ31/GI | Pure Zn coating (6 μm thick) | 40 | ||||
AZ61/GI | Pure Zn coating (6 μm thick) | 70 | ||||
Diffusion brazing | 5 mm AZ31/1 mm 316 L | Lap | Pure Cu (20 μm) | 57 | Mg BM | [80] |
Pure Ni (20 μm) | 32 | Steel/Ni interface | ||||
rTPL bonding | AZ31B/Low carbon steel | Lap (Mg on top) | — | 40 | Interfacial | [44, 81] |
Pure Ag (1 μm) | 201 | Mg BM |
Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by diffusion and eutectic bonding.
Fusion welding involves joining the surface of the materials through melting and solidification to produce the bonding. Fusion-welding technologies based on resistance spot welding [4, 32, 84, 85, 86] laser braze [41, 43, 87, 88, 89, 90, 91, 92], laser hybrid [33, 35, 36, 38, 39, 40, 47, 93, 94, 95, 96, 97, 98] and arc welding [72, 99, 100, 101, 102] have been investigated for joining magnesium alloys to steels. In general, the insertion of an interlayer or mutual diffusion of alloying elements from the BM has been adopted to improve interfacial bonding. The addition of suitable interlayer improves the spreadability and the nucleation of magnesium on steel.
RSW is the most widely used welding technology in the auto industry [103] due its low cost, high speed, ease of operation, and automation [17, 32, 86]. Despite the inherent advantages of RSW, very limited work has been published in joining Mg alloy to steel. This include AZ31B Mg alloy/Zn-coated DP600 steel [32], AZ31 Mg alloy/HDG steel with pre-coated nanoscale Fe2Al5 layer [34], AZ31B-H24 Mg alloy/HDG HSLA steel [4], AZ31B Mg alloy/HDG HSLA steel [84] and AZ31B Mg alloy/electro-galvanized DP600 steel [86].
Most of the works focused on joining magnesium alloys to galvanized steel. Liu and his co-workers developed a novel technique (asymmetric electrode) to lap weld 1.5 mm AZ31B to 1.2 mm HDG DP600 steel successfully by RSW [32]. It was found that the Zn interlayer was squeezed out of the bond region producing deal condition for intimate contact of fresh Mg and steel surfaces. The joining mechanism consisted of braze welding, solid-state bonding, and soldering. Under optimum parameters, a joint with tensile shear strength of 5.0 kN was obtained. Xu et al. [4] compared the microstructure and mechanical performance of AZ31B to HDG HSLA steel joints by RSW and weld-bonding (WB), which combines the RSW and adhesive bonding. It was found that the peak shear load and energy absorption of the weld-bonded magnesium to steel joints were higher than that of resistance spot welded magnesium to steel joints. In another similar study, AZ31B was joined to HDG HSLA steel by RSW under dynamic loading [84]. It was found that the microstructure of the Mg/steel spot welds was different from that of Mg/Mg spot welds, but owing to similar crack propagation and failure mode, both welds had an equivalent fatigue resistance. Interestingly, Feng et al. [86] joined AZ31 to electro-galvanized DP600 steel by RSW process with and without HDG Q235 interlayer. Contrary to the previous studies [4, 32, 84] that used hot-dip Zn coating, the thin and compact features of electro-galvanized zinc layer prevented the zinc-coated layer on the steel to be squeezed out of the nugget. However, with insertion of HDG Q235 interlayer, the zinc-coated layer was squeezed out of the nugget and a peripheral soldered region was formed during the welding process, which significantly improved the joint tensile shear load from 4.14 kN to 5.49 kN. The feasibility of joining AZ31B Mg alloy to DP600 steel via pre-coated nano-scaled Fe2Al5 interlayer was also investigated [34]. The analysis of the interface characteristics revealed that metallurgical bonding was achieved due to the formation of the semi-coherent interfaces of Mg/Fe2Al5/Fe with well matching lattice.
However, RSW of Mg alloy to stainless steel is more challenging because of the absence of any Zn coating. However, Mg alloy to stainless steel joints has been reported [85, 104]. Min et al. [104] investigated the 0.4-mm thick AZ31B sheets, and 0.4-mm thick 443 ferritic stainless steel welded joints using the RSW with 443 ferritic stainless steel cover plates. It was found that Fe-Al IMC layer was formed at the interface, and the molten Mg could wet the surface of the interface layer. However, cracking was observed in the nugget. The crack was associated with the possible thermal behavior of various elements in the nugget during spot-welding process. Recently, Manladan [85] compared the microstructure and mechanical performance of 1.5 mm thick AZ31 Mg alloy/0.7 mm thick 316 L austenitic stainless steel joints by RSW and resistant element welding (REW). In comparison with two-zone FZ, consisting of peripheral FZ on the ASS side and main FZ observed for REW, the RSW joints were produced through welding-brazing mode, in which the Mg alloy melted and spread on the solid steel, forming the nugget only on the Mg side. The RSW produced the weak joint with a peak load of 2.23 kN and energy absorption of 1.14 J, whereas REW produced the strong joint with a peak load of 3.71 kN and energy absorption of 10.2 J.
Generally, in the RSW process of Mg alloys, large electric currents are always needed due to its high electrical and thermal conductivity, which commonly results in expulsion and electrode stick. In addition, RSW created only a localized joint, which may not be particularly strong [105].
A comparison of the Mg alloys/steel joints maximum tensile shear strength produced by RSW is shown in Table 4. It can be seen that limited research has been conducted on magnesium alloys/steel. The presence of zinc layer was crucial for successful welding of magnesium to steel. The predominant failure mode observed is interfacial (IF). This is the kind of failure mode commonly observed when conducting tensile shear tests on spot welds, in which the crack propagates through the nugget. A comparison of the joints performance shows that good static strength has been achieved, almost 95% Mg/Mg spot joint strength by RSW [32]. However, the research on the fatigue performance of Mg/steel dissimilar welds is still at its infancy due to the special geometry of the spot welds; it is hard to predict the crack initiation and propagation rate as reported by Liu et al. [84]. Among the RSW techniques, weld bonding produced the joint with highest mechanical resistance with joint failure on the Mg BM. Weld-bonding joining techniques has the advantages of low manufacturing costs, higher static and fatigue performance, and improved corrosion resistance [106, 107].
Techniques | Materials | Joint design | Transition material | Maximum tensile shear strength (kN) | Failure location | References |
---|---|---|---|---|---|---|
RSW | 1.5 mm AZ31/ 1.2 mm Zn-coated DP600 | Lap (Mg on top) | HDG Zn-coated (9 μm to 12 μm) | 5.0 | Interfacial failure (IF) mode | [32] |
RSW | 1.5 mm AZ31/ 1.2 mm HDG DP600 | Lap (Mg on top) | Bare steel | 0.0 | Not reported | [34] |
Fe2Al5 | 4.8 | Not reported | ||||
RSW | 1.5 mm AZ31B/0.77 mm HSLA steel | Lap (Mg on top) | HDG Zn-coated (11 μm) | 2.0 | Interfacial failure (IF) mode | [84] |
RSW & WB | 2 mm AZ31B-H24/0.7 mm HSLA steel | Lap (Mg on top) | HDG Zn-coated (11 μm) | RSW 4.80 WB 10.0 | IF (RSW) Mg BM (WB) | [4] |
RSW | 2 mm AZ31B/ 1.2 mm Electro-galvanized DP600 | Lap (Mg on top) | No transition material | 4.14 | IF | [86] |
HDG Q235 (0.6 mm) | 5.49 | IF | ||||
RSW & REW | 1.5 mm AZ31/0.7 mm 316 SS | Lap (Mg on top) | No transition material | RSW 2.23 | IF | [85] |
REW 3.71 | PO |
Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by RSW.
Laser beam welding presents a viable option for welding Mg alloys to steel due to its versatility, high specific heat input, and flexibility [55, 108, 109, 110, 111]. Although fusion welding resulted in severe vaporization of Mg alloy at typical welding temperatures, the selection of optimum welding conditions is crucial for successful joining. Therefore, to control the severe vaporization of Mg, considerable number of authors concentrated on laser welding brazing and laser hybrid techniques.
Laser welding brazing (LWB) techniques offered additional advantages such as increased flexibility and adoptability when welding dissimilar metals [28, 91, 92, 112, 113]. To this end, LWB technique is suitable for joining dissimilar materials having large differences in melting points such as Mg/Ti [18, 114, 115, 116], Al/Ti [117], and Al/steel [118, 119]. In particular, welding of magnesium alloys to steel by LWB process was achieved with addition of third material or mutual diffusion of alloying elements. Thin interlayers such as Al [28, 120, 121], Ni [90, 113], Sn [31], Zn [41, 42, 87, 88, 91], and Fe-Al [41], have been used to improve the interfacial bonding between the immiscible Mg and Fe. Miao demonstrated the feasibility of joining 2.4-mm AZ31B Mg alloy to 1.7 mm Q235 steel by laser welding brazing process using high power CO2 laser without welding wire [122, 123, 124]. The analysis of the interface characteristics revealed that transition layer consisted mainly of IMCs, and metal oxides were observed at the interface. The compounds were identified by TEM as Al-rich phases, such as Mg17Al12, Mg2Al3, FeAl, and Fe4Al13 [124]. Although tensile strength of the butt joints could reach 182 MPa (81% of Mg alloy BM), but severe oxidation and vaporization of the magnesium alloy, coupled with immiscibility between magnesium and iron, led to weld defects such as spatters and porosity.
Therefore, to address the severe vaporization of the Mg, laser brazing or laser welding brazing with magnesium-based filler wire was proposed [28]. The benefits of using Al-12Si [28], Ni [113], Sn [31], and Zn [42] as transition material between Mg alloy to steel were explored by Nasiri and co-workers. It was found that the presence of the interlayer significantly improved the spreadability of the liquid magnesium-based filler on steel surface but deteriorated the joint performance because of the weak bonding of IMCs products formed along the interface.
However, several authors observed mutual diffusion of alloying elements when joining Mg alloys to different grades of stainless steel [92, 125, 126]. For instance, [92] compared the joint performance of lap welded 1.5 mm-AZ31B/1.5 mm-mild steel (MS) joints and 1.5 mm-AZ31B/1.5 mm 201 stainless steel (SS) joints produced by LWB. Mechanical bonding was observed for Mg alloy to MS joints, whereas for Mg alloy to SS joints, thin reaction layer identified as FeAl by TEM as shown in Figure 1. The Al diffusion from magnesium BM to the interface was accelerated by chemical potential induced by alloying elements of Cr and Ni was responsible for the interfacial reaction layer obtained.
Mg/201 stainless steel interface characteristics [
A comparison of the Mg alloys/steel joints maximum tensile shear strength produced by LWB is shown in Table 5. The selection of suitable interlayer is crucial for successful bonding. For instance, using Ni and Sn interlayers, which led to formation of the Fe(Ni) and Al8Mn5-Fe(Al) reaction products along the α-Mg-Fe interface, respectively, resulted in formation of strong interfaces with low mismatch strain energy and strong bonds. Therefore, using both Ni and Sn interlayers is recommended for dissimilar joining of steel sheet to magnesium sheet. Furthermore, the diffusion of the alloying elements in BM and the bonding mechanism were extensively studied. A comparison of the joints properties reveals that relatively good static strength of magnesium alloy to steel joints has been obtained [126]. However, the behavior of the Mg alloys/steel joint under dynamic loading is yet to be explored. Among the LWB techniques, dual-beam mode with flux produced joint with an excellent mechanical resistance. To further enhance the reliability of the joint and improve its performance, the possibility of using different interlayers should be explored.
Techniques | Materials | Joint design | Transition material | Maximum strength | Failure mode | References |
---|---|---|---|---|---|---|
Laser welding | 3 mm AZ31B/1.2 mm SP781 steel | Lap (Mg on top) | Zn Coating | 6182 N | AZ31B FZ & Transition zone | [43] |
Laser penetration brazing | 2.4 mm AZ31B/1.7 mm Q235 steel | Butt (0.6 mm offset on Mg) | No interlayer | 182 MPa | Interface | [122, 123, 124] |
Laser brazing | 2 mm AZ31B-H24/1 mm Al-12Si coated steel 2.4-mm- Dia Ti-Braze Mg 600 wire | Single flare bevel Lap (Mg on top) | Al-12 wt-% Si coating (20 ± 2 μm thick) | 767 ± 138 N | Interface | [28] |
Laser brazing | 2 mm AZ31B-H24/1 mm Al-12Si coated steel 2.4-mm- Ti-Braze Mg 600 wire | Single flare bevel Lap (Mg on top) | Pure Ni coating(5 μm) | 1506.3 ± 24.5 N | FZ | [113] |
Laser brazing | 2 mm AZ31B-H24/0.6 mm AISI 1008 | Lap (Mg on top) | Pure Sn coating (3.7 ± 0.7 μm thick) | 2064 ± 85 N | Steel BM | [31] |
Laser brazing | 2 mm AZ31B-H24/0.8 mm AISI 1008 | Lap (Mg on top) | Pure Zn coating (2.6 ± 0.5 μm thick) | 1086.4 ± 150.2 N | Interface | [42] |
Laser welding brazing | 1.5 mm AZ31B-H24/1.5 mm DP980 | Lap (Mg on top) | uncoated | 160 N/mm | Interface | [87, 88, 91] |
Zn coated (10 to 15 μm) | 228 N/mm | |||||
Laser welding brazing | 1.5 mm AZ31B-H24/1.5 mm DP980 with 1.2 mm AZ31 filler | Lap (Mg on top) | uncoated | 190 N/mm | Interface | [41] |
Zn + Fe-Al Phase | 180 N/mm | Interface | ||||
Zn coating (10 μm) | 160 N/mm | Interface | ||||
Fe-Al coating (0.9 μm) | 240 N/mm | Weld Seam | ||||
Laser welding brazing | 1.5 mm AZ31/1.5 mm 22MnB5 with 1.2 mm AZ61 filler | Lap (Mg on top) | AlSi10Fe3-coating | 3090 N | AZ61 filler | [121] |
Laser welding brazing | 1.5 mm AZ31B/1 mm Q235 | Lap (Mg on top) | Al (0.3 mm) | 133 N/mm | FZ | [120] |
Laser welding brazing | 1 mm AZ31B-H24/1 mm Q235A with 1.2 mm AZ92D filler | Lap (Mg on top) | Ni coating (4.5 ± 0.5 μm) | 190 N/mm | FZ | [90] |
Laser Welding brazing | AZ31B/mild Steel | Lap (Mg on top) | No interlayer | 142 N/mm | Interface | [92] |
AZ31B/stainless steel | 270 N/mm | Mg FZ | ||||
Laser welding brazing | 1.5 mm AZ31B-H24/1.5 mm 201 stainless steel and 2 mm AZ31 Filler | Lap (Mg on top) | No interlayer | 274.5 N/mm | Mg HAZ | [125, 126] |
Laser offset welding | 3 mm AZ31/3 mm AISI 316 | Butt (3 mm Offset on Steel) | No interlayer | 100 MPa | AZ31 BM | [30] |
Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by laser welding brazing.
The excessive evaporation of magnesium during fusion welding can be overcome through the method of TIG-assisted laser process [33]. Generally, the addition of TIG could improve absorption of laser power and penetration of molten pool. During laser-TIG process, the TIG torch melts the magnesium, whereas, the laser is used to create deep penetration into the steel [36, 39]. Several authors reported that conventional fusion-welding technology such as TIG or laser welding alone could not be competent for joining of magnesium to steel [33, 89, 94]. For instance, [33, 89] demonstrated during TIG welding of Mg alloy to steel with higher heat input, either the magnesium alloys melt but the steel remained in solid state or the steel melt while blowholes formed in the magnesium alloys. In both cases, the bonding between the immiscible couple could not be obtained. Similarly, for conventional laser-welded magnesium alloys to steel joints, the immiscible materials could be joined with poor weld appearance and poor strength, coupled with large amount of electricity consumed by laser due to low absorptivity of magnesium alloys at room temperature [127]. For TIG-assisted laser process, the reflectivity of the magnesium to laser was reduced and thus the absorption of laser beam improved, creating a deeper penetration in the steel [33, 35, 128]. Consequently, laser-TIG process offers great potential for improving the joint performance for both similar and dissimilar metal welding [33, 39, 40, 89, 129, 130].
Like other joining techniques, interfacial bonding between magnesium alloy to steel could be achieved by insertion of third material or mutual diffusion of alloying elements from the BM [95]. Therefore, the benefits of using Cu [35, 39, 40], Ni [35, 36, 40, 93, 96], Sn [94], Zn [89], Cu-Zn [40, 97] to bond magnesium to steel had been extensively studied. These interlayers formed Mg2X IMC (where X is the interlayer elements) and a solid solution of interlayer in iron along the joint interface, which consequently enhanced the joints mechanical strength [40].
Liu examined the weldability of AZ31B/304 SS by laser-GTA process. A transitional zone consisting of Mg-Fe and magnesium diffusion into the matrix of iron in the form of oxide was observed at the interface. The formation of the complex MgO, ZnO, Fe2O3, or Al2O3 oxides deteriorates the joint strength [39, 94]. Furthermore, the use of laser-TIG techniques to join AZ31B and Q235 steel with Ni, Cu, Sn, and Cu-Zn interlayers was also explored [35, 36, 38, 39, 40, 93, 94]. These interlayers were heated and melted to react with magnesium and steel, and formed a transitional layer in the FZ and solid solution along the steel side. The analysis of the strengthening mechanism revealed that with the addition of suitable interlayers, the joint shear strength could reach a significantly high value or even surpasses Mg alloy BM.
Generally, the corrosion behavior of magnesium alloys is affected by the microstructural variation imposed by welding process [27, 43, 56, 63, 64, 125, 126, 131]. For similar Mg/Mg welds, the grain refinement imposed by welding process was reported to improve the corrosion resistance [132]. However, Liu et al. studied the corrosion behavior of the magnesium to steel joints produced by TIG-assisted laser process in NaCl solution. It was found that the grain refinement and iron splashes imposed by welding process were observed in the weld, which accelerated the corrosion of the magnesium alloy. Interestingly, the use of Al coating was reported to raise the lifespan of the dissimilar joints [47].
A comparison of the Mg alloys/steel joints maximum tensile shear strength produced by laser-TIG process is shown in Table 6. In hybrid laser-TIG welding, excessive vaporization of the magnesium was observed due to the penetration of the laser from upper magnesium plate and the interlayer into the bottom steel. Furthermore, the violent stirring of molten pool restricted metallurgical bonding adjacent to the steel interface, which limited the application of this welding technique. Despite the fast heating and cooling rate of the TIG-assisted laser process, the interfacial reaction was achieved through diffusion and combination of alloying elements from the BM. Based on the existing literature, the interfacial characteristics and the mechanism of wetting in Mg alloys to steel joints produced using TIG-assisted laser process with addition of interlayers were thoroughly investigated. The feasibility of using Cu, Ni, Sn, Zn, and Cu-Zn intermediate elements was also explored. The presence of interlayer was essential for successful joining Mg to steel. The selection of a suitable interlayer was essential for successful bonding. For instance, the Sn added joint shows comparatively lower value due to inhomogeneous compositions in the FZ. A comparison of the joints mechanical properties shows that excellent static strength has been achieved, even surpassing that of magnesium alloy base metal with insertion of Cu and Ni intermediate elements. The high joint shear strength obtained was associated with better wettability and deeper penetration in the weld. However, limited studies focused on corrosion behavior of the TIG-assisted laser magnesium to steel joints. Furthermore, no study has focused on the behavior of the joints under dynamic loading.
Techniques | Materials | Joint design | Transition material | Maximum tensile shear strength (MPa) | Failure mode | References |
---|---|---|---|---|---|---|
Laser-GTA | 1.7 mm AZ31B/1.2 mm 304 steel | Lap (Mg on top) | No interlayer | 90 | Interfacial | [33] |
Laser-TIG | 1.7 mm AZ31B/1.2 mm Q235 | Lap (Mg on top) | No interlayer | 120 | Interfacial | [36, 39, 96] |
Pure Cu foil (0.1 mm thick) | 170 | FZ | [38] | |||
Pure Ni foil (0.1 mm thick) | 166 | |||||
Pure Sn foil (0.1 mm thick) | 117 | |||||
Cu-Zn (H80)-0.1 mm thick | 161 | |||||
Cu-Zn (H62)-0.1 mm thick | 157 | |||||
Laser-TIG | 1.5 mm AZ31B/1.5 mm DP980 | Lap (Mg on top) | Zn coating (10 to 15 μm thick) | 68 | Interfacial | [89] |
Laser-TIG | 1.6 mm AZ31B/1.1 mm Q235 | Butt (0.2 mm offset on steel) | Cu-Zn alloy (H62) 0.1 mm thick | 203 | Weld seam | [97] |
Comparison of the magnesium alloys/steel joints maximum tensile shear strength produced by laser-TIG process.
Arc welding involves joining the materials surface permanently using power supply to obtain an electric arc between the electrode mounted in a torch and a metal. Among the arc welding processes, so far only metal inert gas welding has been used for this material combination.
Metal inert gas (MIG) welding has been widely used in automobile industries due its inherent properties, such as high efficiency, lower cost, and excellent adoptability to material geometry [133, 134]. Therefore, obtaining a reliable Mg alloy to steel joint by MIG welding is essential. However, joining magnesium to steel by MIG is rarely reported, including AZ31B/Zn-coated steel [99, 101, 102, 135], AZ31B/Q235 with Cu interlayer [72, 100] and AZ31B/Aluminized steel [102]. Generally, the addition of suitable interlayer improves the spreadability and the nucleation of magnesium on the steels.
Cold metal transfer (CMT) modified gas metal arc welding of AZ31B Mg to galvanized and bare mild steel sheets showed that welded brazed joints formed in both joints [99, 101].
A comparison of the Mg alloys/steel joints maximum tensile shear strength produced by MIG welding brazing is shown in Table 7. Generally, the investigation on the Mg alloys/steel joints produced by MIG welding is still in its infancy. Therefore, more interlayers should be tested. Moreover, the behavior of the joints under dynamic loading should be studied.
Techniques | Materials | Joint design | Transition material | Maximum tensile strength | Failure mode | References |
---|---|---|---|---|---|---|
CMT Welding | 1 mm AZ31B/Mild steel | Lap (Mg on top) | HDG Zn coating (10 μm) | 224 N/mm | Interfacial | [99, 101] |
Uncoated | 258 N/mm | Mg HAZ | ||||
Bypass current-MIG-welding | 2.5 mm AZ31B/ 2 mm Q235, with 1.6 mm AZ31 filler | Lap (Mg on top) | Zn coating | 133.02 MPa | weld metal | [135] |
MIG spot welding | 1.8 mm AZ31B/1.5 mm Q235 with 1.2 mm ER50-6 filler | Lap (Steel on top) | Pure Cu foil (0.1 mm) | 3200 N | Interfacial | [72] |
MIG spot welding | 3 mm AZ31B/3 mm Q235, 1.6 mm Mg based filler | Butt (V groove) | Uncoated | 160 MPa | Interfacial | [100] |
Pure Cu foil (0.1 mm) | 185 MPa | weld metal | ||||
CMT brazing | 2 mm AZ31/1 mm steel with 1.2 mm AZ31 filler | Lap (Mg on top) | Zn coating (10 μm) | 3100 N | Interfacial | [102] |
Zn-Fe (8 μm) | 4100 N | |||||
Uncoated | 7000 N | Weld metal | ||||
AlSi Coating (25–30 μm) | 6000 N |
Comparison of the Mg alloys/steel joints maximum tensile shear strength produced by MIG welding brazing.
As there is a desire in the aerospace, aircraft, and automotive industries to join magnesium alloys to steel in order to achieve lighter weight, versatile, and tailored properties in one composite part, and development of a welding technology for Mg alloys/steel with a strong metallurgical bond will expedite increased applications of magnesium alloys in these industries. The major challenge in welding of magnesium alloys to steel is the huge differences in physical properties and limited solubility that make the welding process difficult. Different joining processes have been used to join Mg alloys to steel sheets but metallurgical bonding can only be possible with insertion of intermediate interlayer elements or alloy or mutual diffusion of alloying elements from BM. The existence of intermetallic phases or solid solutions between Mg and the interlayer and also the interlayer and Fe is an indication that metallurgical bonding between Mg and Fe using the interlayer may be possible. Formation of thick, brittle intermetallic compounds along the interface between Mg and steel can cause significant deterioration of mechanical properties. Therefore, when choosing the interlayer and the joining process that will be used, minimization of the thickness of any brittle IMCs that might form at magnesium alloy-interlayer-steel joint interfaces and minimization of intermixing between the Mg and Fe in the liquid state are main factors that must be considered.
Solid-state bonding techniques based on FSW, USW, and diffusion and eutectic bonding have been used. Generally, the intimate contact between the dissimilar materials and the formation of IMC with insertion of the suitable interlayer played a significant role in controlling the joint performance of solid-state bonded magnesium alloys to steel.
Joining magnesium alloys to different grades of steels by fusion processes such as RSW, laser welding, and arc welding with and without insertion of interlayers were also reported. Despite the fast heating and cooling rate of the fusion process, the interfacial reaction was achieved through diffusion and combination of alloying elements from the BM. The phase formation in the magnesium-interlayer-steel alloy system and the mechanism of wetting in magnesium-interlayer-steel alloy system were extensibility investigated. The benefits of using Cu, Ni, Sn, Zn, and Cu-Zn interlayers were also explored. The addition of suitable interlayer improves the spreadability and the nucleation of magnesium on the steel substrate. Therefore, more interlayer materials that form the eutectic phase with Mg either in a pure form such as Ag and Al or as alloys such as Al-Cu and Ag-Sn should be investigated.
Currently, a great deal of research has been conducted on the interface characteristics and mechanical performance of Mg alloys to steel joints, particularly under static loading. Under optimized processing conditions, excellent static strength has been achieved, even surpassing that of Mg alloy base metal with insertion of Ag, Cu, and Ni intermediate elements. However, few experiments have been carried out on the corrosion behavior of the jointed parts and the joints performance under dynamic loading. Thus, cost-effective and reliable joining techniques for Mg/steel will still require further development. The need for the industries for more advanced materials to accommodate the huge demands for strong, rigid and light structures may be the powerful drivers for further development of the welding techniques of hybrid structures of Mg alloys to steel.
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This chapter deals with few microbial infections, their association with male infertility, prevention, symptoms, diagnosis, treatment, and control measures.",book:{id:"5059",slug:"genital-infections-and-infertility",title:"Genital Infections and Infertility",fullTitle:"Genital Infections and Infertility"},signatures:"Manonmani Samiappan and Poongothai Jayaramasamy",authors:[{id:"87572",title:"Dr.",name:"Poongothai",middleName:null,surname:"Jayaramasamy",slug:"poongothai-jayaramasamy",fullName:"Poongothai Jayaramasamy"},{id:"185369",title:"Ms.",name:"Manonmani",middleName:null,surname:"Samiappan",slug:"manonmani-samiappan",fullName:"Manonmani Samiappan"}]}],onlineFirstChaptersFilter:{topicId:"1069",limit:6,offset:0},onlineFirstChaptersCollection:[{id:"82174",title:"The Novelty of miRNAs as a Clinical Biomarker for the Management of PCOS",slug:"the-novelty-of-mirnas-as-a-clinical-biomarker-for-the-management-of-pcos",totalDownloads:7,totalDimensionsCites:0,doi:"10.5772/intechopen.104386",abstract:"Polycystic ovary syndrome (PCOS) is a common endocrine disorder that affects around 5–10% of women of reproductive age. The aetiology of PCOS is not fully understood with various genetics, iatrogenic (e.g. chemotherapy) and environmental factors have been proposed. microRNAs (miRNAs) are small non-coding single-stranded RNAs which are known to act as a regulator to gene expression at the post-transcriptional levels. Altered expression of miRNAs has been linked to several disorders including infertility. Recent reports demonstrated the expression of differential levels of miRNAs in the serum, ovarian follicular cells and follicular fluid of PCOS patients when compared with healthy women. Therefore, miRNAs may play important role in the pathogenesis of PCOS. The aim of this chapter is to summarise the current understanding pertaining to miRNAs and PCOS and to expedite its possible role in the diagnosis and management of this disorder.",book:{id:"11085",title:"Polycystic Ovary Syndrome - Functional Investigation and Clinical Application",coverURL:"https://cdn.intechopen.com/books/images_new/11085.jpg"},signatures:"Rana Alhamdan and Juan Hernandez-Medrano"},{id:"82067",title:"Role of Oxidative Stress and Carnitine in PCOS Patients",slug:"role-of-oxidative-stress-and-carnitine-in-pcos-patients",totalDownloads:5,totalDimensionsCites:0,doi:"10.5772/intechopen.104327",abstract:"Polycystic ovary syndrome (PCOS) is a common female endocrine and reproductive system disorder which is found in 6–10% of the female population. PCOS is considered a multifactorial metabolic disease characterized by several clinical manifestations, such as hyperandrogenism, polycystic ovaries and ovulatory dysfunctions. PCOS patients have an increase in the oxidative stress with generation of excessive amounts of reactive oxygen species (ROS) and reduction of antioxidant capacity. Oxidative stress is defined as the imbalance between the production of free radicals and the ability of the organism to defend itself from their harmful effects damaging the plasma membrane, DNA and other cell organelles, inducing apoptosis. Oxidative stress markers are circulating significantly higher in PCOS patients than in healthy women, so these can be considered as potential inducers of the PCOS pathology. Therefore, the central role of the oxidative stress may be involved in the pathophysiology of various clinical disorders including the PCOS. This chapter reviewed the role of oxidative stress and carnitine in PCOS patients, indicating the beneficial action of the carnitine pool, and L-carnitine contributes to restore the energy balance to the oocyte during folliculogenesis and maturation, which represent an important strategy to improve the intraovarian environment and increase the probability of pregnancy.",book:{id:"11085",title:"Polycystic Ovary Syndrome - Functional Investigation and Clinical Application",coverURL:"https://cdn.intechopen.com/books/images_new/11085.jpg"},signatures:"Bassim Alsadi"},{id:"81225",title:"Special Considerations on Hyperandrogenism and Insulin Resistance in Nonobese Polycystic Ovaries Syndrome",slug:"special-considerations-on-hyperandrogenism-and-insulin-resistance-in-nonobese-polycystic-ovaries-syn",totalDownloads:26,totalDimensionsCites:0,doi:"10.5772/intechopen.103808",abstract:"PCOS is a widespread phenotypically inhomogeneous endocrinopathy with significant health consequences and incompletely elucidated pathogenesis. Though visceral adiposity and insulin resistance (IR) is a well-proved pathogenic set of factors of PCOS, not all women with obesity and IR have PCOS and not all PCOS women are obese and have IR, which is explained by certain genetic backgrounds. The reported prevalence of nonobese PCOS (NonObPCOS) is about 20–30%, but it may be higher because especially in lean women with nonclassical phenotypes PCOS diagnosis is often delayed or unrecognized. Unlike obese PCOS, NonObPCOS management is less clear and is limited to symptomatic treatment. This chapter presents in structured fashion the existing results on the prevalence of NonObPCOS, as well as on special aspects of body composition, IR, and hyperandrogenism pathogenesis, including adrenal contribution in NonObPCOS.",book:{id:"11085",title:"Polycystic Ovary Syndrome - Functional Investigation and Clinical Application",coverURL:"https://cdn.intechopen.com/books/images_new/11085.jpg"},signatures:"Tatyana Tatarchuk, Tetiana Tutchenko and Olga Burka"},{id:"80918",title:"Polycystic Ovary Syndrome: It’s Not Just Infertility",slug:"polycystic-ovary-syndrome-it-s-not-just-infertility",totalDownloads:33,totalDimensionsCites:0,doi:"10.5772/intechopen.101923",abstract:"Polycystic ovary syndrome (PCOS) is a heterogeneous endocrine issue described by unpredictable menses, hyperandrogenism, and polycystic ovaries (PCO). The commonness of PCOS changes relying upon which measures are utilized to conclude yet is just about as high as 15–20% when the European culture for human propagation and embryology/American culture for regenerative medication rules are utilized. Clinical signs incorporated grown-ups incorporate sort 1 diabetes, type 2 diabetes, and gestational diabetes. Insulin opposition influences half 70% of ladies with PCOS prompting a few comorbidities including metabolic condition, hypertension, dyslipidemia, glucose narrow-mindedness, and diabetes. Studies show that ladies with PCOS are bound to have expanded coronary corridor calcium scores and expanded carotid intima-media thickness. Psychological wellness problems including despondency, uneasiness, bipolar turmoil, and voraciously consuming food issues additionally happen all the more habitually in ladies with PCOS. Weight reduction works on feminine abnormalities, indications of androgen abundance, and barrenness the board of clinical appearances of PCOS incorporates oral contraceptives for feminine inconsistencies and hirsutism. Spironolactone and finasteride are utilized to treat indications of androgen overabundance.",book:{id:"11085",title:"Polycystic Ovary Syndrome - Functional Investigation and Clinical Application",coverURL:"https://cdn.intechopen.com/books/images_new/11085.jpg"},signatures:"Naheed Akhter, Sadia Sana, Naila Iftikhar, Muhammad Adnan Ahsan, Abu Huraira and Zafaar Siddique"},{id:"80385",title:"Thyroid Dysfunction: In Connection with PCOS",slug:"thyroid-dysfunction-in-connection-with-pcos",totalDownloads:43,totalDimensionsCites:0,doi:"10.5772/intechopen.102492",abstract:"As the prevalence of endocrine dysfunction is increasing and is associated with many complications including polycystic ovary syndrome (PCOS) which, itself is a risk factor of thyroid dysfunction. Although the causality of this association is uncertain, the two conditions share a bidirectional relationship. Both syndromes share certain common characteristics, risk factors and pathophysiological abnormalities, which can be managed by lifestyle changes as well as pharmacological treatment. Polycystic appearing ovaries are a clinical feature of hypothyroidism as well as hyperthyroidism in a few case studies. Adiposity, evidence of deranged autoimmunity, increased insulin resistance and disturbed leptin levels are present in both the disease states, seeming to play a complex role in connecting these two disorders. Major endocrine pathways including hypothalamic-pituitary-thyroid axis (HPTA) and HP-gonadal axis are involved in parallel relationship of PCOS and thyroid dysfunction. This chapter helps to explore all the dimensions of the relationship between PCOS and thyroid dysfunction.",book:{id:"11085",title:"Polycystic Ovary Syndrome - Functional Investigation and Clinical Application",coverURL:"https://cdn.intechopen.com/books/images_new/11085.jpg"},signatures:"Mariya Anwaar and Qaiser Jabeen"},{id:"80033",title:"Novel Methods in the Diagnosis of PCOS: The Role of 3D Ultrasonographic Modalities",slug:"novel-methods-in-the-diagnosis-of-pcos-the-role-of-3d-ultrasonographic-modalities",totalDownloads:62,totalDimensionsCites:1,doi:"10.5772/intechopen.101995",abstract:"Polycystic ovary syndrome (PCOS) is a common and complicated endocrine disorder, with its diagnosis based on clinical, laboratory and imaging criteria. The latter is usually assessed via two-dimensional ultrasound; however, the advent of three-dimensional ultrasound, along with three-dimensional power Doppler (3D-PD) could offer more accurate diagnoses and further our understanding of PCOS pathophysiology. Three-dimensional ultrasound (3D-US) has already been used successfully in many fields of gynecology. It offers improved image quality with stored data that can be processed either manually or automatically to assess many parameters useful in PCOS assessment, such as ovarian volume, number of follicles and vascular indices. The examination requires minimal time as data is assessed in post-processing, thus being more tolerable for the patient. 3D-US parameters are generally increased in PCOS patients when compared to controls and 2D measurements, with studies showing improved diagnostic performance, though that remains inconclusive. 3D transrectal ultrasound is more accurate in the diagnosis of virgin PCOS patients than the modalities currently available in that subgroup. Overall, though with some limitations, 3D-US is a promising diagnostic method in the assessment of PCOS which, regardless of diagnostic accuracy, can undoubtedly offer many practical advantages, more objective and reliable measurements, potentially improving PCOS diagnosis standardization.",book:{id:"11085",title:"Polycystic Ovary Syndrome - Functional Investigation and Clinical Application",coverURL:"https://cdn.intechopen.com/books/images_new/11085.jpg"},signatures:"Apostolos Ziogas, Emmanouil Xydias and Elias Tsakos"}],onlineFirstChaptersTotal:9},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:0,limit:8,total:null},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:90,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:104,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:32,numberOfPublishedChapters:320,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:12,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:141,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:133,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:113,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:107,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:5,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:17,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}},{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. 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",coverUrl:"https://cdn.intechopen.com/series/covers/22.jpg",latestPublicationDate:"June 27th, 2022",hasOnlineFirst:!0,numberOfPublishedBooks:1,editor:{id:"356540",title:"Prof.",name:"Taufiq",middleName:null,surname:"Choudhry",slug:"taufiq-choudhry",fullName:"Taufiq Choudhry",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000036X2hvQAC/Profile_Picture_2022-03-14T08:58:03.jpg",biography:"Prof. Choudhry holds a BSc degree in Economics from the University of Iowa, as well as a Masters and Ph.D. in Applied Economics from Clemson University, USA. In January 2006, he became a Professor of Finance at the University of Southampton Business School. He was previously a Professor of Finance at the University of Bradford Management School. He has over 80 articles published in international finance and economics journals. His research interests and specialties include financial econometrics, financial economics, international economics and finance, housing markets, financial markets, among others.",institutionString:null,institution:{name:"University of Southampton",institutionURL:null,country:{name:"United Kingdom"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:3,paginationItems:[{id:"86",title:"Business and Management",coverUrl:"https://cdn.intechopen.com/series_topics/covers/86.jpg",isOpenForSubmission:!0,editor:{id:"128342",title:"Prof.",name:"Vito",middleName:null,surname:"Bobek",slug:"vito-bobek",fullName:"Vito Bobek",profilePictureURL:"https://mts.intechopen.com/storage/users/128342/images/system/128342.jpg",biography:"Dr. Vito Bobek works as an international management professor at the University of Applied Sciences FH Joanneum, Graz, Austria. He has published more than 400 works in his academic career and visited twenty-two universities worldwide as a visiting professor. Dr. Bobek is a member of the editorial boards of six international journals and a member of the Strategic Council of the Minister of Foreign Affairs of the Republic of Slovenia. He has a long history in academia, consulting, and entrepreneurship. His own consulting firm, Palemid, has managed twenty significant projects, such as Cooperation Program Interreg V-A (Slovenia-Austria) and Capacity Building for the Serbian Chamber of Enforcement Agents. He has also participated in many international projects in Italy, Germany, Great Britain, the United States, Spain, Turkey, France, Romania, Croatia, Montenegro, Malaysia, and China. Dr. Bobek is also a co-founder of the Academy of Regional Management in Slovenia.",institutionString:"Universities of Applied Sciences FH Joanneum, Austria",institution:null},editorTwo:{id:"293992",title:"Dr.",name:"Tatjana",middleName:null,surname:"Horvat",slug:"tatjana-horvat",fullName:"Tatjana Horvat",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002hXb0hQAC/Profile_Picture_1642419002203",biography:"Tatjana Horvat works as a professor for accountant and auditing at the University of Primorska, Slovenia. She is a Certified State Internal Auditor (licensed by Ministry of Finance RS) and Certified Internal Auditor for Business Sector and Certified accountant (licensed by Slovenian Institute of Auditors). At the Ministry of Justice of Slovenia, she is a member of examination boards for court expert candidates and judicial appraisers in the following areas: economy/finance, valuation of companies, banking, and forensic investigation of economic operations/accounting. At the leading business newspaper Finance in Slovenia (Swedish ownership), she is the editor and head of the area for business, finance, tax-related articles, and educational programs.",institutionString:null,institution:{name:"University of Primorska",institutionURL:null,country:{name:"Slovenia"}}},editorThree:null},{id:"87",title:"Economics",coverUrl:"https://cdn.intechopen.com/series_topics/covers/87.jpg",isOpenForSubmission:!0,editor:{id:"327730",title:"Prof.",name:"Jaime",middleName:null,surname:"Ortiz",slug:"jaime-ortiz",fullName:"Jaime Ortiz",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00002zaOKZQA2/Profile_Picture_1642145584421",biography:"Dr. Jaime Ortiz holds degrees from Chile, the Netherlands, and the United States. He has held tenured faculty, distinguished professorship, and executive leadership appointments in several universities around the world. Dr. Ortiz has previously worked for international organizations and non-government entities in economic and business matters, and he has university-wide globalization engagement in more than thirty-six countries. He has advised, among others, the United Nations Development Program, Inter-American Development Bank, Organization of American States, Pre-investment Organization of Latin America and the Caribbean, Technical Cooperation of the Suisse Government, and the World Bank. Dr. Ortiz is the author, co-author, or editor of books, book chapters, textbooks, research monographs and technical reports, and refereed journal articles. He is listed in Who’s Who in the World, Who’s Who in America, Who’s Who in Finance and Business, Who’s Who in Business Higher Education, Who’s Who in American Education, and Who’s Who Directory of Economists. Dr. Ortiz has been a Fulbright Scholar and an MSI Leadership Fellow with the W.K. Kellogg Foundation. His teaching interests revolve around global economies and markets while his research focuses on topics related to development and growth, global business decisions, and the economics of technical innovation.",institutionString:null,institution:{name:"University of Houston",institutionURL:null,country:{name:"United States of America"}}},editorTwo:null,editorThree:null},{id:"88",title:"Marketing",coverUrl:"https://cdn.intechopen.com/series_topics/covers/88.jpg",isOpenForSubmission:!0,editor:{id:"203609",title:"Associate Prof.",name:"Hanna",middleName:null,surname:"Gorska-Warsewicz",slug:"hanna-gorska-warsewicz",fullName:"Hanna Gorska-Warsewicz",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bSD9pQAG/Profile_Picture_2022-06-14T11:58:32.jpeg",biography:"Hanna Górska-Warsewicz, Ph.D. is Associate Professor at Warsaw University of Life Sciences and Head of Department of Food Market and Consumption Research. She specializes in the subject of brands, brand equity, and brand management in production, service, and trade enterprises. She combines this subject with marketing and marketing management in both theoretical and practical aspects. Prof. Hanna Górska-Warsewicz also analyzes brands in the context of trademarks, legal regulations and the protection of intangible. She is an author or co-author of over 200 publications in this field, including 8 books. 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Empirical Evidence from an Emerging Economy",doi:"10.5772/intechopen.105589",signatures:"Bishnu Kumar Adhikary and Ranjan Kumar Mitra",slug:"does-board-structure-matter-in-csr-spending-of-commercial-banks-empirical-evidence-from-an-emerging-",totalDownloads:9,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Corporate Social Responsibility",coverURL:"https://cdn.intechopen.com/books/images_new/11602.jpg",subseries:{id:"86",title:"Business and Management"}}},{id:"82395",title:"Toward a Better Understanding of Green Human Resource Management’s Impact on Green Competitive Advantage: A Conceptual Model",doi:"10.5772/intechopen.105528",signatures:"Hosna Hossari and Kaoutar Elfahli",slug:"toward-a-better-understanding-of-green-human-resource-management-s-impact-on-green-competitive-advan",totalDownloads:13,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Corporate Social Responsibility",coverURL:"https://cdn.intechopen.com/books/images_new/11602.jpg",subseries:{id:"86",title:"Business and Management"}}},{id:"82269",title:"CSR Reporting and Blockchain Technology",doi:"10.5772/intechopen.105512",signatures:"Pattarake Sarajoti, Pattanaporn Chatjuthamard, Suwongrat Papangkorn and Piyachart Phiromswad",slug:"csr-reporting-and-blockchain-technology",totalDownloads:9,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Corporate Social Responsibility",coverURL:"https://cdn.intechopen.com/books/images_new/11602.jpg",subseries:{id:"86",title:"Business and Management"}}}]},overviewPagePublishedBooks:{paginationCount:1,paginationItems:[{type:"book",id:"11392",title:"Leadership in a Changing World",subtitle:"A Multidimensional Perspective",coverURL:"https://cdn.intechopen.com/books/images_new/11392.jpg",slug:"leadership-in-a-changing-world-a-multidimensional-perspective",publishedDate:"May 11th 2022",editedByType:"Edited by",bookSignature:"Muhammad Mohiuddin, Bilal Khalid, Md. 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