Optical fiber sensors with a microhole produced by femtosecond laser drilling.
\\n\\n
Dr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\\n\\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\\n\\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\\n\\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\\n\\nThank you all for being part of the journey. 5,000 times thank you!
\\n\\nNow with 5,000 titles available Open Access, which one will you read next?
\\n\\nRead, share and download for free: https://www.intechopen.com/books
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Preparation of Space Experiments edited by international leading expert Dr. Vladimir Pletser, Director of Space Training Operations at Blue Abyss is the 5,000th Open Access book published by IntechOpen and our milestone publication!
\n\n"This book presents some of the current trends in space microgravity research. The eleven chapters introduce various facets of space research in physical sciences, human physiology and technology developed using the microgravity environment not only to improve our fundamental understanding in these domains but also to adapt this new knowledge for application on earth." says the editor. Listen what else Dr. Pletser has to say...
\n\n\n\nDr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\n\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\n\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\n\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\n\nThank you all for being part of the journey. 5,000 times thank you!
\n\nNow with 5,000 titles available Open Access, which one will you read next?
\n\nRead, share and download for free: https://www.intechopen.com/books
\n\n\n\n
\n'}],latestNews:[{slug:"stanford-university-identifies-top-2-scientists-over-1-000-are-intechopen-authors-and-editors-20210122",title:"Stanford University Identifies Top 2% Scientists, Over 1,000 are IntechOpen Authors and Editors"},{slug:"intechopen-authors-included-in-the-highly-cited-researchers-list-for-2020-20210121",title:"IntechOpen Authors Included in the Highly Cited Researchers List for 2020"},{slug:"intechopen-maintains-position-as-the-world-s-largest-oa-book-publisher-20201218",title:"IntechOpen Maintains Position as the World’s Largest OA Book Publisher"},{slug:"all-intechopen-books-available-on-perlego-20201215",title:"All IntechOpen Books Available on Perlego"},{slug:"oiv-awards-recognizes-intechopen-s-editors-20201127",title:"OIV Awards Recognizes IntechOpen's Editors"},{slug:"intechopen-joins-crossref-s-initiative-for-open-abstracts-i4oa-to-boost-the-discovery-of-research-20201005",title:"IntechOpen joins Crossref's Initiative for Open Abstracts (I4OA) to Boost the Discovery of Research"},{slug:"intechopen-hits-milestone-5-000-open-access-books-published-20200908",title:"IntechOpen hits milestone: 5,000 Open Access books published!"},{slug:"intechopen-books-hosted-on-the-mathworks-book-program-20200819",title:"IntechOpen Books Hosted on the MathWorks Book Program"}]},book:{item:{type:"book",id:"5209",leadTitle:null,fullTitle:"River Basin Management",title:"River Basin Management",subtitle:null,reviewType:"peer-reviewed",abstract:"Many of the challenges facing farmers and human communities from hydrographical basins are not new. But, due to the fact that the nature and extents of the problems vary from one region to another and from one basin to another, the responses are highly diversified. There is no generally valid solution for all the problems. However, in addressing issues on ensuring the prevention or mitigation of the destructive consequences of flood damage or prolonged drought as for the optimal use of water by consumers, sustainable basin land use, biodiversity conservation, and environment protection as well, fourteen specialists and their colleagues present the state of the art in these important matters and new possible solutions to solve, identified from the scientific investigations undertaken.",isbn:"978-953-51-2605-8",printIsbn:"978-953-51-2604-1",pdfIsbn:"978-953-51-6666-5",doi:"10.5772/61557",price:139,priceEur:155,priceUsd:179,slug:"river-basin-management",numberOfPages:314,isOpenForSubmission:!1,isInWos:1,hash:"09b5a27ccab9d67afa66f2f0a14fb1a4",bookSignature:"Daniel Bucur",publishedDate:"August 10th 2016",coverURL:"https://cdn.intechopen.com/books/images_new/5209.jpg",numberOfDownloads:13964,numberOfWosCitations:8,numberOfCrossrefCitations:12,numberOfDimensionsCitations:21,hasAltmetrics:0,numberOfTotalCitations:41,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 20th 2015",dateEndSecondStepPublish:"November 10th 2015",dateEndThirdStepPublish:"February 14th 2016",dateEndFourthStepPublish:"May 14th 2016",dateEndFifthStepPublish:"July 27th 2016",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6,7",editedByType:"Edited by",kuFlag:!1,editors:[{id:"50794",title:"Prof.",name:"Daniel",middleName:"G",surname:"Bucur",slug:"daniel-bucur",fullName:"Daniel Bucur",profilePictureURL:"https://mts.intechopen.com/storage/users/50794/images/system/50794.jfif",biography:"Daniel Bucur is currently a professor of Land Improvement at the Pedotechnics Department, University of Applied Life Sciences and Environment in Iasi, Romania.\r\nHe completed his doctorate at the Technical University of Iasi in 1998.\r\nHis major research areas include water excess removal, irrigation, soil erosion control, climate changes, and sustainable land management. In recent years, he has been in charge of many national and international research projects, including Soil Erosion and Conservation Measures, Effect of Sewage Sludge Application on Quality Indices of Soil Vulnerable to Degradation, Sustainable Development of Soil Resources from the Areas with Drainage Works, and Impact of the Hydro-climatic and Pedo-geomorphological Risks on the Environment in Small Catchment.\r\nHe has published more than 150 papers in reviewed journals, 5 book chapters, and 9 books apart from more than 30 unreviewed papers and reports.",institutionString:"University of Applied Life Sciences and Environment in Iasi",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"5",totalChapterViews:"0",totalEditedBooks:"3",institution:{name:"University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca",institutionURL:null,country:{name:"Romania"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"788",title:"Hydropower Engineering",slug:"hydropower-engineering"}],chapters:[{id:"50930",title:"Landslides Caused by Climate Change and Groundwater Movement in Permafrost Mountain",doi:"10.5772/63068",slug:"landslides-caused-by-climate-change-and-groundwater-movement-in-permafrost-mountain",totalDownloads:1065,totalCrossrefCites:1,totalDimensionsCites:1,signatures:"Wei Shan, Ying Guo, Zhaoguang Hu, Chunjiao Wang and\nChengcheng Zhang",downloadPdfUrl:"/chapter/pdf-download/50930",previewPdfUrl:"/chapter/pdf-preview/50930",authors:[{id:"139091",title:"Dr.",name:"Wei",surname:"Shan",slug:"wei-shan",fullName:"Wei Shan"},{id:"162871",title:"Dr.",name:"Ying",surname:"Guo",slug:"ying-guo",fullName:"Ying Guo"},{id:"184953",title:"Mr.",name:"Zhaoguang",surname:"Hu",slug:"zhaoguang-hu",fullName:"Zhaoguang Hu"}],corrections:null},{id:"51200",title:"River Basin Management in the Past and at Present and its Impact on Extreme Hydrological Events",doi:"10.5772/63398",slug:"river-basin-management-in-the-past-and-at-present-and-its-impact-on-extreme-hydrological-events",totalDownloads:877,totalCrossrefCites:1,totalDimensionsCites:2,signatures:"Jana Moravcová, Václav Bystřický, Jiří Pečenka, Jakub Polenský,\nTomáš Pavlíček, Nikola Nováková and Pavel Ondr",downloadPdfUrl:"/chapter/pdf-download/51200",previewPdfUrl:"/chapter/pdf-preview/51200",authors:[{id:"181059",title:"Ph.D.",name:"Jana",surname:"Moravcová",slug:"jana-moravcova",fullName:"Jana Moravcová"},{id:"181579",title:"Mr.",name:"Pavel",surname:"Ondr",slug:"pavel-ondr",fullName:"Pavel Ondr"},{id:"181580",title:"Mr.",name:"Jiri",surname:"Pecenka",slug:"jiri-pecenka",fullName:"Jiri Pecenka"},{id:"181581",title:"Mr.",name:"Jakub",surname:"Polensky",slug:"jakub-polensky",fullName:"Jakub Polensky"},{id:"181583",title:"Mr.",name:"Vaclav",surname:"Bystricky",slug:"vaclav-bystricky",fullName:"Vaclav Bystricky"},{id:"185438",title:"Mr.",name:"Tomas",surname:"Pavlicek",slug:"tomas-pavlicek",fullName:"Tomas Pavlicek"}],corrections:null},{id:"50558",title:"Streamflow Response to Climate Variability and Land‐Cover Changes in the River Beça Watershed, Northern Portugal",doi:"10.5772/63079",slug:"streamflow-response-to-climate-variability-and-land-cover-changes-in-the-river-be-a-watershed-northe",totalDownloads:971,totalCrossrefCites:2,totalDimensionsCites:2,signatures:"Adélia. 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\r\n\tThe use of dyes and pigments is narrowly associated with everyday life. Since ancient times, people have been using various types of dyes and pigments for both aesthetic and practical applications. Typically, the coloration of various materials e.g. textiles, clay, plastics, etc. has been their main purpose. Yet, the scope of contemporary dyes and pigments has become significantly broader and there is constant interest in new products fulfilling numerous requirements parallel to their ability to act as colorants. This trend has led to the development of functional dyes.
\r\n\r\n\tIn recent years, novel dyes and pigments with hi-tech applications have been developed and there is a continuous demand for new products with better properties and/or broader application scope. Of particular interest is the development of dyes and pigments with environment-responsive aptitudes i.e. products that can undergo some structural modification as a result of external stimuli e.g. light, heat, pressure, pH-changes, etc. These stimuli-responsive functional dyes have in turn found application in sensor technologies, optical data storage, molecular switches, etc. Acknowledging these facts, this book aims to cover current state-of-the-art research and development in the remarkably important area of environment-responsive (multi)functional dyes and pigments.
",isbn:"978-1-83968-615-3",printIsbn:"978-1-83968-614-6",pdfIsbn:"978-1-83968-616-0",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"624f533946a159bc8a03f109c2e1dc91",bookSignature:"Dr. Raffaello Papadakis",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10203.jpg",keywords:"Fluorescent Dyes, PH-Sensitive Dyes, Solvatochromism, Solvent Polarity Indicators, Chromic Betaines, Viscosity, Charge-Transfer Complexes, Spectroscopy, Piezochromism, Optoelectronics, Photochromism, Molecular Switches",numberOfDownloads:62,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"August 28th 2020",dateEndSecondStepPublish:"September 25th 2020",dateEndThirdStepPublish:"November 24th 2020",dateEndFourthStepPublish:"February 12th 2021",dateEndFifthStepPublish:"April 13th 2021",remainingDaysToSecondStep:"5 months",secondStepPassed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:"Chemical Engineer with working experience at the Institute of Molecular Sciences, CNRS/Aix-Marseille University in the research group of Dr. Thierry Tron, and at Uppsala University in the research group of Dr. Henrik Ottosson. Currently a Senior Research Scientist at Tdb Labs, Uppsala, Sweden.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"251885",title:"Dr.",name:"Raffaello",middleName:null,surname:"Papadakis",slug:"raffaello-papadakis",fullName:"Raffaello Papadakis",profilePictureURL:"https://mts.intechopen.com/storage/users/251885/images/system/251885.jpg",biography:"Raffaello Papadakis is a Chemical Engineer (MEng 2005) majoring in organic chemical technology and polymer science and technology. He started his PhD in the field of physical organic chemistry in 2006 under the supervision of Prof. (Emer.) Dr. Athanase Tsolomitis (National Technical University of Athens, Greece) and graduated in 2010. During his PhD he concentrated on the synthesis of solvatochromic probes and molecular switches. He later on spent two years in Marseille, France (September 2010–January 2013) working as a postodoctoral researcher at the Institute of Molecular Sciences, CNRS/Aix-Marseille University, in the field of water oxidation catalysts in the research group of Dr. Thierry Tron before moving to to Uppsala, Sweden in 2014. There he joined the group of Dr. Henrik Ottosson (Uppsala University) and he worked as a postdoc researcher and later as a researcher (Forskare) focusing on excited state (anti)aromaticity and graphene photochemistry-related research. His current research interests revolve around physical organic and materials chemistry with an emphasis on the chemistry and photochemistry of graphene and novel covalent organic frameworks as well as polymer chemistry. He is the author and coauthor of 24 scientific research papers, two book chapters and he has more than 35 contributions in international conference proceedings. Furthermore, he is an active referee of scientific peer-reviewed papers of world-class chemistry journals and he has acted as a scientific expert evaluating international research-grant proposals. Currently he works as a Senior Research Scientist at TdB Labs AB (Sweden) and specializes in polysaccharide modifications and derivatization placing particular interest in fluorescent dye polysaccharide-functionalizations.",institutionString:"TdB Labs",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"2",totalChapterViews:"0",totalEditedBooks:"0",institution:null}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"8",title:"Chemistry",slug:"chemistry"}],chapters:[{id:"74730",title:"Treatment of Textile Dyeing Waste Water Using TiO2/Zn Electrode by Spray Pyrolysis in Electrocoagulation Process",slug:"treatment-of-textile-dyeing-waste-water-using-tio2-zn-electrode-by-spray-pyrolysis-in-electrocoagula",totalDownloads:63,totalCrossrefCites:0,authors:[null]}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"247865",firstName:"Jasna",lastName:"Bozic",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/247865/images/7225_n.jpg",email:"jasna.b@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, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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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:"878",title:"Phytochemicals",subtitle:"A Global Perspective of Their Role in Nutrition and Health",isOpenForSubmission:!1,hash:"ec77671f63975ef2d16192897deb6835",slug:"phytochemicals-a-global-perspective-of-their-role-in-nutrition-and-health",bookSignature:"Venketeshwer Rao",coverURL:"https://cdn.intechopen.com/books/images_new/878.jpg",editedByType:"Edited by",editors:[{id:"82663",title:"Dr.",name:"Venketeshwer",surname:"Rao",slug:"venketeshwer-rao",fullName:"Venketeshwer Rao"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"4816",title:"Face Recognition",subtitle:null,isOpenForSubmission:!1,hash:"146063b5359146b7718ea86bad47c8eb",slug:"face_recognition",bookSignature:"Kresimir Delac and Mislav Grgic",coverURL:"https://cdn.intechopen.com/books/images_new/4816.jpg",editedByType:"Edited by",editors:[{id:"528",title:"Dr.",name:"Kresimir",surname:"Delac",slug:"kresimir-delac",fullName:"Kresimir Delac"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3621",title:"Silver Nanoparticles",subtitle:null,isOpenForSubmission:!1,hash:null,slug:"silver-nanoparticles",bookSignature:"David Pozo Perez",coverURL:"https://cdn.intechopen.com/books/images_new/3621.jpg",editedByType:"Edited by",editors:[{id:"6667",title:"Dr.",name:"David",surname:"Pozo",slug:"david-pozo",fullName:"David Pozo"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"52487",title:"Femtosecond Laser-Induced Surface Modification and its Application",doi:"10.5772/65428",slug:"femtosecond-laser-induced-surface-modification-and-its-application",body:'\nUltrashort pulse laser interaction has been attractively applied to the modification of material properties [1–8] in various materials. The ultrashort pulse laser applications to micromachining [9–16] have received much attention. This is mainly because of two reasons that are extremely high-peak power and ultrashort pulse less than material thermal relaxation. In the high-peak laser regime, multiphoton absorption can be easily induced even in transparent materials. Furthermore, the effects of thermal diffusion during material processing can be extremely minimized with ultrashort pulses that produce a thermal non-equilibrium state between electrons and lattice. Owing to such non-thermal and spatially localized effects can facilitate to locate a specified volume ablation without collateral thermal damage, and microdevices can be precisely machined using femtosecond processing to produce high functionality even in a microscale platform such as thin optical fibers [17–25] or analytical microchip for biochemical application. For this reason, such processing for functional microstructures has been previously reported in the past decade.
\nUsing femtosecond laser micromachining for such optical devices, surface characteristics should be carefully checked because the optical/mechanical properties on the laser-induced surface structures provide completely different response with dependence on micro/nanoscale conditions which is varied with laser irradiation parameters. Such surface responses can be used for controlling phenomena like wettability, reflectivity, and friction as well as biomimetics [26]. This is the reason why nanotechnology has attracted wide interest in many fields.
\nIn this chapter, we present femtosecond laser micromachining to microdevice such as fiber optics with a focus on surface qualities. Section 2 is dedicated to laser-induced periodic surface structures (LIPSSs), which are induced in low- and high-fluence regime. The influences of laser-induced structures for the fabrication of fiber-optic sensors are described in Section 3, with experimental techniques and results in our research group. Conclusions are presented in Section 4.
\nLaser-induced surface structures have been widely investigated in various materials such as metals, ceramics, glass, and polymers. A periodicity of about the wavelength λ of the laser radiation is commonly observed, which are usually called as ripples or laser-induced periodic surface structures (LIPSS). It should be noted that such LIPSS show water-repellent properties [12, 27–29], diffusive reflection [30–34], and friction change [35–38]. The spatial periodicity of low-spatial frequency LIPSS (LSFL) is known to be approximately laser wavelength dependent, and the LSFL is mainly orientated perpendicular to the direction of the laser polarization. LSFL and high-spatial frequency LIPSS (HSFL) generated by femtosecond laser pulses have been recently observed. Using high-energy laser pulses, micrometer-size-rugged surfaces are formed inside the channel structures ablated by femtosecond laser pulses [39–41]. It is also important to understand the formation mechanism of micrometer-scale surfaces for femtosecond laser direct writing technologies in which relatively higher laser energies are used to fabricate a 3D structure such as fluidic channels [3, 42, 43], sampling cells [19, 21, 22], and interferometers [44–46]. Different generating processes will induce different mechanical responses, which will be detailed in the following.
\nIn 1965, LIPSS formation induced by exposing semiconductors to a ruby laser was first observed by [47]. Although LIPSS had been unfavorable as irregular structures inevitably accompanied by laser processing, the development of ultrashort pulse laser allows us to achieve not only stable periodic structures, but also non-conventional advantages based on nanotechnologies mentioned above. Applications of LIPSS for mechanical engineering such as water-repellent property, strain relief [48], and friction control have been reported as well as for biomedical applications such as cell adhesion and its growth [49, 50]. For the case of optics, LIPSS are applied to structural coloring [51], surface-enhanced Raman spectroscopy (SERS) [52], and anti-reflection surface. Mechanism of LIPSS formation has been discussed in many literatures. Sipe et al. established a first principle for LIPSS formation process, by considering a surface-scattered wave from the surface roughness and modeling the effect of the roughness on the electromagnetic field [53–55]. Figure 1 shows a simple model for the formation of LIPSS, showing geometry of a laser beam idealized as a finite plane wave onto a rough surface. While LSFL can be obtained by either a CW (continuous wave) laser or a pulsed laser irradiation, HSFL can only be observed for the case of using pulse laser in the duration of picosecond or femtosecond range. For linearly polarized light at a normal direction to the plane, an LIPSS periodicity becomes much smaller than the laser wavelength and their direction can be parallel [56] or orthogonal [57] to the polarization, depending on the material properties and the irradiation parameters. The nature of HSFL is still under controversially discussion, and many theories have been proposed to explain the formation mechanism: surface-plasmon polaritons [58–60], second harmonic generation [57, 61], self-organization [62–67], waveguides modes [68], parametric process [69, 70], or interference along with a modification of the optical properties during the pulse [56].
\nGeometry of a one-dimensional rough surface of length L. The medium below the rough interface is electrically defined by the permittivity ε and permeability µ. The surface roughness is assumed to be confined in a “selvedge region” in a range between z = 0 and z = 1, where 1 ≪ λ.
Discussion of a contribution of surface plasmon polaritons has been presented by Bonse et al. [71, 72] and Miyaji et al. [58, 59], where the formation of LSFL and HSFL was experimentally observed with the change of optical properties induced by an electron excitation under laser irradiation. Huang et al. showed that self-organized LSFL on dielectric materials was formed via a metallic-like behavior by an interference of initial surface plasmon and laser pulses followed by the grating assisted surface plasmon-laser coupling [67]. They also obtained a good agreement with their experimental results, by taking account into the effect of surface plasmon. Reif et al. reported that HSFL is due to self-organization structure formation during the relaxation of highly non-equilibrium condition after explosive ion emission [63]. Straub et al. demonstrated that generation of a dense electron-hole plasma at a carrier concentration slightly beyond the critical plasma density allows for the excitation of surface plasma waves at the high-spatial frequencies that are required for HSFL formation [60]. Bonse et al. explained HSFL formation based on the interference between the surface-scattered wave of the laser and its second harmonic [57, 71, 73]. Okumuro et al. found that the periodicity of HSFL can be explained by induction of a surface-plasma wave through the parametric process of laser light [69]. They also showed that laser fluence dependence of periodic grating structures was formed on metal surfaces under femtosecond laser pulse irradiation. The results indicate that the formation of periodic grating structures depends not only on target properties but also on the electron density of plasma produced in the femtosecond excitation regime [70].
\nFemtosecond laser machining with high fluence can selectively remove large volumes of material only in the focal area. Thermal effect could be observed in higher fluence more than the threshold value of materials even though heat diffusion outside the focal area is comparatively minimized in contrast to longer pulse lasers. Several literatures have provided evidence for the existence of a molten region surrounding ablation area, even for the ultrashort pulse processing [39–41, 74]. Thermocapillary force and hydrodynamic force will be dominated in the high-fluence regime. Theoretical model for the regime has also been fabricated and seems to be in good agreement with experimental results [41]. Yaker et al. showed reliable evidence regarding the formation mechanism of surface microstructure induced by high-fluence femtosecond pulses. They tried to fabricate a microflow channel for microfluidic devices by scanning femtosecond laser beam on a borosilicate glass. The experiment was performed by mutually overlapping thin rims surrounding the laser-induced smooth crater to produce surface microstructures as shown in Figure 2.
\n\nThe crater surrounded by thin rim is created by resolidification of a molten layer generated during the ablation process. They found that a thin rim is formed around the craters and is created on the undamaged surface. To investigate the mechanism of granulated rim formation by successive laser pulses, they also demonstrated theoretical analysis of the thermal and fluid processes taking into account ablation process in the femtosecond regime. The series of the experiment indicate that a thin molten layer below the surface and the rim is generated by the high pressure resulted from a dense plasma producing a pressure-driven fluid motion of the molten material outwards from the center of laser focal point as shown in Figures 2 and 3. The second laser pulse is diffracted by the rim created by the first laser pulse, resulting in diffraction pattern on the crater area overlapped. The distance between the diffraction rims seems to be approximately equal to the wavelength of the laser light, which means that diffraction by the rim plays an important role for the formation of granulated structures. Flow forces applied to a molten layer are generated from expanding plasma which can be happened by thermocapillary forces and forces driven by plasma expansion above the irradiated surface. There are a number of processes in the regime of femtosecond pulse-induced phenomena, including nonlinear absorption, plasmas, shock propagation, melt propagation, and resolidification. These processes can roughly be broken down into three different time domains as illustrated in Figure 4.
\nA schematic of ablation process with a femtosecond laser pulse in the characteristic time (a)–(c) of excitation and relaxation process.
Femtosecond laser-induced crater rim and its diffraction pattern generated by overlapping two pulses. (a), (c) One laser pulse. (b), (d) Two overlapping pulses. (SEM pictures adapted with permission from Ref. [41], © IOP Publishing. Reproduced by permission of IOP Publishing. All rights reserved).
Characteristic time of excitation and relaxation process.
The physics and discussions mentioned in this section are quite important for the case of using femtosecond laser microstructuring into microdevices such as a thin optical fiber to be highly functionalized. The following sections show the functionalization of optical fibers using femtosecond laser drilling, the operation of which requires a large number of pulses, and high fluence much larger than a material threshold to fabricate 3D microstructures. The surface-granulated structures found in the above approach are also discussed with recent experimental results in our research groups.
\nIn the past three decades, fiber-optic sensor technology has been developed for different applications. The progresses in research have developed the fiber-optic sensors for practical applications. For instance, distributed fiber-optic sensors have been installed in dams and bridges to monitor the performance of these facilities. With the rapid growth of optical networks, the cost of fiber-optic sensors has significantly decreased as key components in fiber-optic sensors (such as optical fiber, light sources, and photo detectors) are commercially available in optical communications industries. Therefore, it can be anticipated that fiber-optic sensors will be used in more extensive applications in sensing technology. Recent interest in the technologies has not only been focused on the development of novel ideas but also on technologies for integrating high-performance devices of various functionalities onto a single optical fiber, which are sometimes referred to as “lab-on-fiber” technologies. The authors have previously reported a new approach to the development of a fiber-optic spectrometer by embedding a microhole to be a spectroscopic sample cell in multimode optical fiber, using a near-ultraviolet (NUV) femtosecond laser [22].
\nOptical fiber sensors machined by femtosecond laser drilling are summarized in Table 1. The fiber-optic sensor with a microhole working as a sensing area in single-mode optical fibers [19, 20] was demonstrated to monitor the refractive index (RI) of liquid, in which the microhole structures were fabricated by femtosecond laser irradiation of near infrared (NIR) at 800 nm with a 120-fs duration pulse at 1 kHz repetition rate during 15 s and pulse energy of 11 µJ. The sensor performances seem to be limited for introducing liquid into the microhole when using high viscous liquids because the shapes of microholes are sharply tapered over the entire hole due to objective lens used to focus laser beam and its fabrication procedure of one side laser irradiation. The other experiment [21] was conducted by using 1030-nm NIR femtosecond laser with a 280-fs pulse of pulse energy of 1–5 µJ and 100-kHz high repetition rate during 5 min, and immersion oil is also required to reduce the laser-induced thermal energy. An optical fiber was immersed in water to avoid the thermal influence, where helical drilling method was also employed in the fabrication procedure in order to reduce tapering the microhole. However, the tapering in the lateral direction of optical axis still remains at the opposite side of cladding because laser beam is focused by passing through the cylindrical body of the optical fiber processed. For the case of femtosecond laser deep drilling to dielectrics, laser energy can be nonlinearly transferred to electrons via multiphoton absorption, where the order of N-photon ionization process is determined by photon energy. This fact indicates that ablation threshold becomes lower with decreasing wavelength [75, 76] because efficient energy transfer to electrons can be easily achieved via a shorter-wavelength femtosecond laser via efficient multiphoton absorption with reducing extra energy diffusion by collision cascade [77, 78]. Consequently, it can be supposed that a shorter-wavelength laser with a higher photon energy is comparatively favorable for microstructuring in microscale platforms such as thin optical fibers because the shorter-wavelength beam can reduce thermal debris inevitably generated through resolidification of a molten material. Such debris disturbs growth of microhole by absorbing/scattering subsequent laser pulses and redeposition of debris on the hole surface, resulting in poor surface quality of the inner wall. Actually, the experiment conducted on the fabrication of microhole with 1030-nm femtosecond laser in air atmosphere was tested experimentally and failed [21]. Moreover, it was found that the surface microstructures on the inner wall are remarkably different between using NIR and NUV femtosecond laser drilling. Further advantage of use of 400-nm laser deep drilling is found in the fact that Rayleigh range can be longer. In contrast to the hole drilling using NIR wavelength (e.g. 800 nm [19, 20], 1030 nm [21]) which were inevitably accompanied with debris and longer processing period more than 10 s or 5 min, efficient deep hole drilling in glass optical fiber was successfully demonstrated with reducing debris in a very short irradiation time of approximately 1.0 s to fabricate a fiber-optic inline spectrometers, using a 400-nm, 1-kHz, and 350-fs laser irradiation.
\nLaser and its irradiation condition | \nCell | \n|||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
References | \nPulse width (fs) | \nWavelength (nm) | \nRepet-ition (kHz) | \nPulse energy | \nIrradi- ation time | \nFabric- ation process | \nLens NA | \nNumber of hole | \nShape (taper angle) | \nHole diameter (µm) | \nRoughness | \nVolume (single cell) (pL) | \n
[19] | \n120 | \n800 | \n1 | \n11 µJ | \n5 s | \nSingle direction | \n0.25 | \n1 | \nDead-end (7°–10°) | \n6 | \n– | \n1.2 | \n
10 s | \n8 | \n6.3 | \n||||||||||
15 s | \n11 | \n12 | \n||||||||||
[20] | \n– | \n– | \n1 | \nThrough hole (4°–18°) | \n15 | \n>3 µm | \n60–80 | \n|||||
[21] | \n280 | \n1030 | \n100 | \n1–5 µJ | \n5 min | \nHelical drilling | \n0.68 | \nSM: 1 MM: 3 | \nThrough hole (1°–4°) | \n20 | \n300 nm (no evidence) | \n40 | \n
[22] | \n350 | \n400 | \n1 | \n15 µJ | \n1.2 s | \nTwo direction | \n0.40 | \nMM: 1, 10 | \nThrough hole (3.7°) | \n10 | \n<500 nm | \n20 | \n
Optical fiber sensors with a microhole produced by femtosecond laser drilling.
The following sections show results obtained in our recent study regarding the fabrication of optical fiber sensors using femtosecond laser with high-fluence regimes. As shown in Figures 5 and 6, the microholes are intentionally designed and machined to easily guide a liquid sample into microhole working as a sensing area without immersion liquids during fabrication process. The sensor part has microhole array penetrating through the whole fiber core by drilling from both sides of the fiber cladding in order to avoid tapering the opening apertures inlet/outlet of through hole.
\n\nThe microholes were fabricated with optical train, as illustrated in Figure 5. A multimode optical fiber (MMF, core diameter 62.5 µm) was irradiated with femtosecond pulses from two directions (not simultaneously) to avoid tapering of the inlet/outlet of the cell. The fundamental of a 1 kHz Ti-sapphire laser (IFRIT Cyber Laser Inc., 1 mJ/pulse, λ = 800 nm, 210 fs pulse duration) was converted to the second harmonic (240 µJ/pulse, 350 fs pulse duration) in a wavelength converter [second harmonic generation (SHG) unit, manufactured by Cyber Laser Inc.]. The second harmonic pulses were introduced to an objective lens through an optical train to guide and focus the beam at the target optical fiber. The collimating optics reduce the beam diameter from 6.0 to 2.8 mm so that a longer Rayleigh distance compared with the initial beam can be obtained using a negative and positive lens array. This prevents plasma generation in the air between the lens combination, which is important because the laser beam could be defocused and the energy could be lost by plasma creation during collimation and thereby affected by diffraction before the final focusing. An optical fiber irradiated was mounted on a three-dimensional motor-controlled translation stage equipped with a rotary mechanism for rotation of the fiber. The laser focal point was carefully adjusted by moving the translation stage in the X, Y, Z, and θx directions. The focal point, defined as the center of the Rayleigh distance, was set to be 5 μm beyond the cladding surface of the inlet.
\nThe cross-sectional view in vicinity of the focal point and two different types of collimations.
Figure 6(a) and (b) indicates the schematic images of side and cross-sectional views of the through holes embedded in optical fiber lines, respectively. Based on the microscopic image showing the fabricated through hole (c), the hole diameters were found to be approximately 18 and 10 µm at the opening aperture and waist, respectively, the volume of which was calculated to 19.8 pL by assuming that its shape could be approximated by two truncated cones with taking into account the taper angle of the microhole. Optical micrographs of the through hole of the side and top view are presented in Figure 6(c) and (d), respectively, showing that the microholes are connected together so as to produce a through hole. Taking a look at the inside of the through hole by using scanning electron microscope (SEM) as shown in Figure 6(e), a morphology modification was formed to be granulated structure, which have been reported so far in the previous works as mentioned above in the femtosecond regime [41]. The sample of Figure 6(e) was prepared in such a way that the inner surface of a part of hole was exposed by intentionally cleaving the fabricated fiber along an off-axis plane of the through hole to clearly see the rugged surface with SEM. The rough surface could be generated with folded debris or microrims surrounding laser-induced craters by successive laser pulses. Importantly, the size of the rugged particle is found to be approximately a few hundred nanometers. Such rough surface gives more diffusive reflection on the hole boundary, hence higher optical scattering, especially for shorter-wavelength light wave. Additionally, it should also be noted that such particle-like structure will be more water repellent to a liquid by which water inside becomes easier to flow. As shown in Table 1, the surface roughness should be depended on the fabrication method. The shorter wavelength laser irradiation can reduce the increasing thermal debris and its redeposition on the inner surface, the fact of which can be seen from the comparison between Refs. [20] and [22].
\nThe schematic drawings of microholes in a fiber-optic line monitored from side view (a) and the cross section (b). The photographs of (c) and (d) give examples which show the shape of the through hole and the hole opening, respectively. (e) SEM picture of a part of hole. Adapted with permission from Ref. [22].
To evaluate the intrusion/discharge velocity of liquids sucked into/drained out from a through hole, the real-time response of optical intensity change was measured in such a way that the sensing part with a single through hole is alternatively immersed in water and ethanol as indicated in Figure 7. Sample liquids were immediately sucked into microholes by capillary driving force and as was firstly confirmed by monitoring the optical intensity of transmitting light and staining the microhole with a color dye simultaneously. As soon as the sensing part was lifted from a liquid pool, the liquid held in the microhole seems to be immediately drained out in a few seconds because of the hydrophobic-repellent structure.
\n\nFigure 8 shows transmitting spectra to investigate sensing response for RI changes in a wavelength between 400 and 1000 nm. In this measurement, two types of sensor samples which had (a) a single cell and (b) 10 cells were prepared to compare the spectroscopic measurement between them. Taking a look at Figure 8(b), remarkable decreases in transmission can be confirmed in a shorter wavelength range (λ < 600 nm), because light transmitted through the fiber core was scattered at the hole surfaces by the fact that the wavelength of transmitted light becomes approximately equal to the scale of surface roughness. By reducing or controlling the surface roughness on the inner wall, sensing performances of fiber optics with an inline sample cell in terms of sensitivity, reproducibility, response time, and recovery time as well as versatility for sensing target could be improved for practical use. High-quality surface finish therefore is required for the improvement of the sensing performances even in high-fluence regimes. The roughness with a periodicity of <100 nm could be useful for reducing the scattering in a shorter wavelength range, by roughly estimating the reflectivity on the inner wall based on Mie-scattering effect.
\n(a) Experimental setup to measure intrusion/discharge velocity of liquids. (b) The real-time response of optical intensity change measured by alternatively immersing in water and ethanol. (c) and (d) show one of a response waveform using ethanol and water, respectively.
According to the experimental data [19–22] (summarized in Table 1), the roughness might be reduced by immersing an optical fiber in water or immersion oils even though the fabrication period becomes unfavorably much longer. Instead of using liquid immersion processing, we have been trying to reduce the roughness using a NUV 400-nm femtosecond laser so as to achieve a high-quality inner surface of microhole in a high-throughput fashion. Figure 9 shows SEM pictures of the inner surface of a dead-end microhole fabricated in a flat silica glass plate. The experiment was performed to make a smooth surface on the inner wall of the hole with varying laser irradiation parameters. As can be seen from Figure 9(a), wave-like nanostructures with a spatial periodicity of 100–200 nm are observed on the bottom of the hole. Figure 9(b) and (c), respectively, shows the inner surface of the hole at different depths near and far from the hole inlet. Surface roughness is still observed at the periphery of the inlet of Figure 9(b) because the surface roughness could be worse as the number of irradiated pulses becomes greater. On the other hand, the case of the inner surface Figure 8(c) seems to be much smoother than the surface (b). To figure out the formation mechanism of the granulated surface in a high-energy regime, further experiment was carried out on this point.
\nTransmission spectra for the case of sensor samples (a) single cell and (b) 10 cells when immersion in three types of liquids. Adapted with permission from Ref. [22].
The surface morphology of shallow craters was generated by single and multiple pulses with an emphasis on rims surrounding the craters. In this experiment, shallow craters were formed on a fused silica plate by overlapping focused laser pulses on the same spot as shown in Figure 10. The shape of the crater created in this experiment is depended on incident laser beam, the pattern of which is deformed through a second harmonic generator (SHG unit manufactured by Cyber Laser Inc.). A crater surrounded by a single rim was observed by SEM analysis of Figure 11(b). The average thickness of the first rim is approximately 500 nm. Figure 11(c) shows an overlapped crater created by two pulses. Molten materials are driven away from the crater to almost 10 µm distance. The second image (Figure 11(d)) shows that a new rim is formed inside the first one by overlapping a second pulse. The distance between the two rims is approximately equal to the wavelength of the irradiated laser beam. We also found a belt-shape structure on a part of the rim. The composition and formation mechanism still need to be explored. For the case of Figure 11(f): overlapping three pulses, it was interestingly found that the height of third rim is comparatively low simultaneously accompanied with nanofibers [79–81] with a diameter ranging from a few to few tens of nanometers. It should be noted that, instead of the growth of the rim, nanofibers were grown by only a few pulses. In order to obtain optimum surface conditions for the fiber-optic inline spectrometer, further investigations are in progress to improve the surface finish.
\nSEM images of a microhole generated on a flat silica glass plate, by successive laser pulses of the energy of 20 µJ and using focusing optics (inserted table (b) in Figure 5). (a) The whole fabricated microhole at 5000× magnification and higher resolution SEM pictures at hole bottom (b), the side wall near the hole inlet (c), and the side wall near the bottom (d).
The schematic image of crater rims produced by overlapping focused laser pulses on the same spot. (a) One laser pulse. (b) Two overlapping pulses.
SEM images of shallow craters with thin rim generated on a flat silica glass plate with varying the number of pulses. The whole crater at 5000× magnification (a, c, e) and higher resolution (b, d, f).
In this chapter, the authors presented an overview of laser-induced periodic surface structures (LIPSS) produced by ultrashort laser pulses. A part of its historical background was provided; the formation mechanism of LIPSS in femtosecond pulse regimes was described. Experimental results obtained by our studies were also presented as a practical use of femtosecond micromachining even though a part of the result is still under investigation.
\nSection 4 shows the functionalization of optical fibers using femtosecond laser drilling. High-fluence femtosecond laser microstructuring was described with recent experimental data in our research group. The investigation not only to explain the formation mechanism of LIPSS but also to have a microscale perspective should be important for material removal because a high-throughput fabrication of 3D microstructures to microdevices requires a large number of pulses and high fluence. We therefore expect that the obtained results can contribute to practical aspects.
\nThis work has been partially supported by JSPS KAKENHI Grant Number 15K04735.
\nWilson Disease (WD) is an autosomal recessive genetic metabolic disease of copper metabolism. Its incidence is vary in different geographic areas with an average incidence of 1 in 30,000 individuals worldwide. Recent studies suggest a considerably higher prevalence of 1:1500–1:3000 for WD. It is caused by mutations in the ATP7B gene encoding a copper transporting P-type ATPase required for copper excretion into the bile [1, 2]. WD is first described by the American neurologist Samuel Alexander Kinnier Wilson in 1912. There are earlier case reports mostly by neurologist in mid 1800s [3]. Kayser and Fleischer mentioned the pigmented corneal rings, in 1902 and 1903 respectively In 1911, Wilson presented his monograph describing the “progressive lenticular degeneration”. Bramwell, in 1916, was the first to realize the importance of liver pathology in WD. In 1948, Cumings described the copper abnormalities in WD and in 1952, Scheinberg and Gitlin showed that the ceruloplasmin levels were low in most of WD patients. In 1956, Walshe introduced the penicillamine as a chelating agent, the first effective treatment for the condition [3, 4]. This discovery of successful chelation therapy makes WD one of the most satisfying genetic diseases to be diagnosed and treated.
Originally WD was described as a neurodegenerative disease associated with cirrhosis of the liver. Later, WD was observed in children and adolescents with acute or chronic liver disease without any neurologic symptoms [5]. Now, WD is considered a multi-systemic disorder, in which hepatic, neurological and psychiatric symptoms are often associated with renal, endocrine, osteoarticular, corneal and myocardial disturbances, all related to abnormal copper metabolism ending with systemic accumulation of the copper [6, 7].
Ultrastructural findings of WD have also been studied. The mitochondrial changes are the most distinctive and pathogenetically significant and include heterogeneity of size and shape, increased matrix density, separation of inner from outer membranes, enlarged intercristal spaces and various types of inclusions. Importantly, ultrastructural mitochondrial changes in WD cannot be considered pathognomonic; although exceedingly rare with cholestatic liver disease, such changes are found with mtDNA depletion disorders [4, 8].
WD has considerable variation in clinical presentations, the most common ones being liver disease and neuropsychiatric disturbances [9]. There is considerable phenotypic variation in WD: Some patients present with hepatic disease during the first decade of life, some with neurological degeneration in adolescence or adult life, with or without overt liver disease. In a study by Ferenci et al., the severity of liver disease did not show correlation with the mutation status. Rather, they reported that the prevalence of cirrhosis increased with age in pediatric patients. They found that hepatic disease was more common among females, whereas neurological presentation occurred more frequently in males [10]. The wide range of disease patterns cannot be explained just by different mutations. Environmental, epigenetic, and other genetic factors are also contribute pathogenesis of WD [6, 10].
Classically low serum copper and low ceruloplasmin levels with high urinary copper content make a triad which is usually associated with WD diagnosis. But this triad may be absent or incomplete in 3% of genetically confirmed WD cases [7].
Early diagnosis of WD is important. But it is also important to make the diagnosis of WD prior to transplantation. Because organ transplant networks make special provision for acute liver failure (ALF) due to WD when considering the urgency of transplantation and the terminology relating to acute presentations of liver disease become relevant when listing a case of WD [11].
A large variability in the age of onset and in the clinical presentation of WD exists. Hepatic manifestations of WD at presentation can be extremely variable, and range from asymptomatic hepatomegaly, isolated splenomegaly, persistent or intermittent elevation of serum aminotransferases, jaundice, fatty liver or pseudo-autoimmune hepatitis, acute hepatitis, compensated or decompensated cirrhosis to acute liver failure (ALF). The varied clinical manifestations of WD due to pathological copper accumulation in different organs, even in the early course of the disease, often pose a diagnostic challenge [7].
The main therapeutic strategy is using chelating agents, particularly D-penicillamine. Liver transplantation (LT) is reserved for patients unresponsive to medical therapy or with fulminant hepatic failure. LT for neurological complications is highly controversial and generally cannot be recommended [8].
Copper is an essential element for cellular function. Dietary copper is absorbed in the stomach and duodenum and reaches the liver by the portal vein [1]. Intestinal uptake is regulated by the Menkes ATPase (ATP7A). The ATP7A gene is expressed in most tissues except the liver. Menkes disease, an X-linked copper deficiency disorder, results from mutations in this gene. The abnormal gene in Wilson disease is ATP7B (the Wilson ATPase) which shows 56% homology to ATP7A [8]. It is expressed mainly in the liver but its expression is not restricted to liver cells. This data suggests that ATP7B dysfunction might be responsible for the systemic disturbances of copper trafficking in the whole human body [1, 6]. The hepatic protein ATP7B encodes a copper-transporting P-type ATPase, transporting copper into the secretory pathway for incorporation into apoceruloplasmin, forming ceruloplasmin. ATP7B moves copper into the trans-Golgi network, where ceruloplasmin peptide acquires its complement of copper, assumes its folded state and is then released into the circulation [12]. Excess is excreted eventually into the bile. Without the normal complement of copper, the peptide folds differently and has a decreased circulating half-life, leading to a low level of serum ceruloplasmin. Biliary excretion of copper is necessary for its homeostasis. When ATPB7 is defective, excess copper accumulates in the hepatocytes. Eventually the excess copper exceeds the storage capacity causing hepatocellular injury and release of copper into the circulation. Most WD patients have a low level of circulating ceruloplasmin which is a direct result of defective copper handling in hepatocytes as a result of mutation of the ATP7B gene. Free copper is extremely toxic and can produce irreversible cellular damage. The functional consequences of pathogenic ATP7B mutation are increased intracellular copper levels. This produces oxidative stress and free radical formation as well as mitochondrial dysfunction, which results in cell death in the liver, brain, kidneys, heart, eyes, and joints. As this disease damages multiple systems at one time, it poses a diagnostic challenge [2]. Over 600 gene alteration in ATP7B were recognized [6, 12]. The most common ones are single-nucleotide missense and nonsense mutations, chased by insertions/deletions, and, rarely, splice site mutations. H1069Q is the most common mutation around the world, seen in most of the WD carriers in Europe and USA, with some absence for this mutation in some countries [6]. Correlation of phenotype with specific mutations (genotype) is difficult in Wilson disease because the vast majority of affected individuals are compound heterozygotes, possessing one copy each of two different mutations. Differences in clinical features of various mutations between siblings and even identical twins suggests that other genes or environmental factors are important [6, 8]. In a study by Ferenci et al., it was suggested that the HSD17B13:TA allele may modulate the phenotype and outcome of WD by reducing the transition from copper induced hemolysis to fulminant WD. Furthermore, it is associated with milder histological changes [10]. When testing for mutations of the WD gene ATP7B becomes inexpensive and rapid, genetic testing may become the starting point for diagnostic investigation [1].
WD has a myriad of clinical presentations, hepatic, neurological, ophthalmic and psychiatric, that mimic other conditions. WD may present at any age. Although most patients present between ages 5 and 35, the age range is much wider. There are cases reported as early as 9 months and as late as the eighth decade [1, 2, 13]. So far, the oldest patient in English literature is a 77- year-old Turkish woman [14].
Approximately one half of the patients with WD present with liver disease. In the majority of cases, WD manifests its presence during childhood or teenage years in the form of liver symptoms [7]. Hepatic symptoms and presentations of WD are very variable from asymptomatic cases to cases with overt cirrhosis or with ALF. Liver disease may mimic all forms of common liver conditions. All children with an apparent diagnosis of autoimmune hepatitis should also be investigated for WD, and adults with a presumptive diagnosis of autoimmune hepatitis failing to respond rapidly and appropriately to corticosteroid therapy must be carefully evaluated for WD. In terms of the rate of progression of the disease, cirrhosis is usually diagnosed in the second decade of life, although some individuals do not develop cirrhosis, even after the fourth decade of life [15]. Hepatic manifestations usually present earlier than neurological symptoms by 5 years. The most common hepatic signs and symptoms are jaundice, hepatomegaly and abdominal pain [1]. In a subset of patients focal liver lesions may show up, showing with a wide run of imaging highlights. The lion’s share of focal liver lesions in patients with WD are benign nodules, but there are reports that have depicted malignant liver tumors or dysplastic nodules in these patients. Although rare in WD compared to other liver diseases, hepatocellular carcinoma occurs in patients of all ages. Cholangiocarcinoma may also occur in WD [8].
Neurologic manifestations include tremor, gait disturbances, choreiform movements, Parkinsonism or akinetic rigid syndrome i.e., partial parkinsonism, dysarthria, pseudobulbar palsy, rigid dystonia, seizures, migraine headaches, and insomnia. In WD cohorts, neurological presentation is associated with a significantly longer time from onset of symptoms to diagnosis than hepatic presentation, ranging from 2.5 to 6 years. In large case series, mean age at onset of neurologic problems extends from 15 to 21 a long time of age, a decade after onset of liver disease, but a number of patients have been analyzed with a starting neurologic onset earlier than age 10 [7]. Psychiatric manifestations encompass depression, neuroses, personality changes, psychosis and poor performance at school. It was reported that 30—40% of patients have psychiatric symptoms at diagnosis and 20% had seen a psychiatrist prior to their WD diagnosis [12]. WD should be ruled out in any teenager with unexplained cognitive, psychiatric, or movement disorder [13]. Neuropsychiatric signs are the predominant presentation in adults but also may be present in up to 50% of teenagers. WD should also be included in the differential diagnosis work-up of unclear neuropsychiatric syndromes in patients after age 60 years [9].
Ocular findings include the Kayser–Fleischer (KF) ring, due to copper accumulation in Desçemet’s membrane, and sunflower cataracts, due to copper accumulation in the lens. They are diagnosed by slit lamp examination. In known cases of hepatic WD, the rings are present in just over half of patients. KF rings are usually absent in children with liver disease. KF rings are rarely observed in other conditions such as in patients with chronic cholestatic diseases, monoclonal gammopathies, multiple myeloma, arci senilis, and pulmonary carcinoma and are thus not specific for WD [1, 7]. Of note, KF rings are not so easy to diagnose without experience, some authors suggest that anterior segment Scheimpflug imaging (Pentacam, Oculus) could be more helpful to diagnose or confirm KF rings by ophthalmologists with little experience in patients with WD. In general it is said that when neurological symptoms are present, KF rings is present in almost all WD patients at disease diagnosis [7]. But there are reports of cases with neurological involvement without KF rings [8].
Other presentations and clinical findings are intermittent bouts of jaundice caused by haemolysis, gynaecomastia, amenorrhoea, repeated spontaneous abortion, cardiac complications including ECG abnormalities, ventricular fibrillation, cardiomyopathy, orthostatic hypotension, urolithiasis, renal tubular disease, hypoparathyroidism, pancreatitis and rhabdomyolysis [8].
Elucidation of some straightforward biochemical tests have been appeared to be both touchy and decently particular for WD. Two such records incorporate a proportion of alanine aminotransferase (ALT) by aspartate aminotransferase (AST), and a proportion of alkaline phosphatase (ALP) by total bilirubin (TB). An ALT/AST proportion of more than 2.2 contains a sensitivity of 94% and a specificity of 86%; the ALP/TB proportion of less than 4 encompasses a sensitivity of 94% and a specificity of 96% [4].
In Wilson disease, the 24-hour urine copper excretion is usually >100 μg (1.6 μmol) and almost always exceeds 40 μg (0.6 μmol). When penicillamine 500 mg is administered by mouth at the beginning and 12 h later during a 24-hour urine collection, copper excretion >25 μmoles (1587 μg) per 24 h is taken as diagnostic. This test has been validated only in children, and its sensitivity is not as great as originally thought [8].
Ceruloplasmin is the major carrier for copper in the blood. Testing for serum ceruloplasmin is often done when searching for the cause of unexplained liver disease. There are physiologic variations in the serum level of ceruloplasmin. It is very low in early infancy to the age of 6 months, peak at higher than adult levels in early childhood, and then decrease to the normal adult range [1]. A serum ceruloplasmin level < 200 mg/L (<20 mg/dL) has been considered consistent with WD, and diagnostic if associated with KF rings. Except WD, conditions such as marked renal or enteric protein loss, severe end stage liver disease of any etiology, neurologic diseases copper deficiency, and Menkes disease can show low ceruloplasmin levels [1, 7, 13].
Total serum copper (which incorporates non–ceruloplasmin bound copper or “free copper” and copper joined in ceruloplasmin) is ordinarily diminished in extent to the diminished serum ceruloplasmin. However, in patients with WD with extreme liver damage, serum copper may be inside the ordinary extend or uniquely hoisted within the setting of ALF due to the discharge of copper from liver tissue stores and the increase in free copper in the blood [13]. A novel approach is the direct specification of labile copper (non-Cp-bound copper), called interchangeable copper (CuEXC). It permits to calculate the “relative replaceable copper” (REC) which alludes to the proportion of CuEXC to total copper. REC was assessed as a convenient diagnostic appliance for WD with a high sensitivity and specifity allows the calculation of relative interchangeable copper (REC) that compares to the proportion between CuEXC and total serum copper. It is represented that REC is a great diagnostic biomarker with a specifity and specificity near to 100% for the determination of WD when its value is >18.5%. It allows a separation of Wilsonian liver disease from other types of liver disorders such as autoimmune, infectious. Moreover, REC can make a great aid to family screening, because it is possible to make a distinction between WD patients and heterozygous carriers or healthy subjects. The CuEXC value at diagnosis indicates of extrahepatic involvement and its seriousness [7]. But further studies are needed to evaluate its diagnostic accuracy in children with liver disease [13].
The urine copper shows to the sum of non-ceruloplasmin bound copper within the circulation. Urinary copper concentration is measured per 24 h since there’s noteworthy changeability within the copper substance of spot urine collections for them to be utilized. The customary level taken as demonstrative of WD is >100 μg/24 h (>1.6 μmol/24 h) in symptomatic patients [1]. In asymptomatic children or children with mild liver disease, urinary copper values are often normal [13]. However, high urinary copper values may be seen in other sorts of liver disorders (e.g., autoimmune hepatitis, unremitting active liver disease, or cholestasis and in specific during acute liver failure of any etiology). Heterozygotes may too have borderline levels [7].
The diagnosis is not fundamentally straightforward indeed even when the disease is effectively being considered. In a patient within the age-range 5–50 years who has liver disease or characteristic neurological symptoms, finding serum caeruloplasmin underneath 5 mg/dL is profoundly compatible with WD; association too a Kayser–Fleischer (KF) ring affirms the diagnosis. In nearly one-third of patients, serum caeruloplasmin can be within normal limits. As a sole, serum caeruloplasmin is not an adequate diagnostic test for WD. KF rings are diagnostic, but they can also be seen in patients who have persistent cholestasis of other etiology. Lack of KF rings happens in around 50% of adult patients with liver disease and hence does not run the show out WD. KF rings may not be determined even when there’s neurological involvement.
Liver biopsy is typically performed when clinical and laboratory findings are not diagnostic or for evaluation of unexplained liver disease or abnormal liver tests. Another aim is to determine the degree of hepatic inflammation and for hepatic copper quantitation [1]. The spectrum of hepatic pathological changes occurring in WD is very broad, ranging from elementary changes typical of a toxic pathology, to inflammatory changes typical of viral or autoimmune etiology [6]. The main features are microvesicular and macrovesicular steatosis, glycogenated hepatocyte nuclei, inflammation, and variable hepatocellular anisonucleosis [16, 17].
The manifestations of liver involvement have a varied spectrum depending on the stage of the disease. In the earlier steps, hepatocyte injury may at first manifest as simple steatosis (Figure 1) with frequent association of glycogenated nuclei. Steatosis, Mallory-Denk bodies (MBDs), lipogranulomas and glycogenated nuclei have been represented as characteristic morphologic findings in liver biopsies with WD. This picture frequently imitates alcoholic and non-alcoholic fatty liver disease [6]. The distinction from nonalcoholic steatohepatitis (NASH) depends upon the demonstration of accumulated copper in the hepatocytes by histochemical stains. Lipofuscin accumulates in periportal areas, and some of the granules are large, irregular in shape and vacuolated. The intermediate stage of the disease shows histological features similar to those of chronic hepatitis of any etiology including viral or autoimmune hepatitis, with the arrival of the portal and periportal inflammation composed of lymphocytes and plasma cells, which results in the destruction of the limiting plate, and parenchymal necrosis followed by bridging fibrosis [4]. Because of low-titer autoantibodies (mainly antinuclear antibodies) are commonly found in patients with WD, differential diagnosis with autoimmune hepatitis (AIH) can be more complicated. Also, cases of WD and concomitant AIH have been reported [13]. More than 50% of cases may show the presence of intra-cytoplasmic eosinophilic MBDs (Figure 2). The literature suggests that steatosis, glycogenated nuclei and MBDs in periportal hepatocytes are features that may be used to distinguish the chronic hepatitis of WD from other more common etiologies [1]. In the cirrhotic stage which is usually macronodular but can be mixed or even micronodular, the histologic features are non-specific, and usually little or no inflammation is present. Some cases may show mild steatosis or features of steatohepatitis. Clusters of large hepatocytes with a granular eosinophilic cytoplasm (oncocytic or oxyphil cells), resulting from an increased number of mitochondria, are often seen but this is not specific for WD [8]. The distribution of copper is quite variable, with some of the cirrhotic nodules containing a lot and others containing little or none. Defining widespread copper deposits by histochemistry can help for the diagnosis. It should be noted that the distribution of copper is variable: some nodules with prominent staining, others with minimal or none (Figure 3). This could generate false negative impression in biopsy specimens, and it has been suggested that two liver cores may be needed for copper detection and diagnosis. Cases which present with ALF or fulminant hepatitis, the histology includes portal and parenchymal inflammatory infiltrate, associated with hepatocyte injury, swelling and necrosis. There may be massive or submassive necrosis. Copper can be demonstrated in hepatocytes and when there has been significant necrosis, in Kupffer cells and portal macrophages [1, 8]. In contrast, copper is rarely demonstrable in Kupffer cells or portal macrophages in the cirrhotic stage [8].
Steatosis and anisonucleosis in a hepatectomy specimen (H&E).
Mallory-Denk bodies in a hepatectomy specimen (H&E).
Heterogenous copper accumulation in a hepatectomy specimen (Rhodanine).
Excess copper storage in the hepatocytes is a relevant sign of WD, and determination of hepatic copper content in the liver biopsy, is important in the diagnosis of WD. This may be accomplished by utilizing special histochemical stains for copper which are rhodanine, rubeanic acid and Timm’s silver stains, and for copper related protein of which are orcein, aldehyde fuchsin and Victoria blue. None of these stains is fully sensitive nor specific. Orcein reveals the accumulation of metallothioneins, the proteins involved in excess copper sequestration. Positive staining appears as large irregular granules dark-brown in color. In the Timm’s stained slides, if there is mild accumulation copper shows up small black or greenish-black granules in the intracytoplasmic perinuclear area or canalicular side of hepatocytes, and when there is heavy accumulation, the whole cytoplasm of the hepatocyte stuffed with coarse granules. With rhodanine stain copper accumulation appears as small red granules [1, 6, 8]. Out of granular staining, diffuse cytoplasmic staining pattern (Figure 4) can be seen with copper stains, which is frequently reported in WD [18].
Diffuse cytoplasmic staining pattern (Timm).
The most effective method is vary in different reports. In our study with transplant hepatectomies, we found that positivity rates of Timm, rhodanine and orcein are 85%, 82%, and 48% respectively (Figure 5A-C). We thought that pannodular (prominent diffuse staining of nodule), staining is a powerfull indicator of WD. In this context, we suggested that pannodular staining is a more convincing staining pattern for the histopathologic diagnosis of WD and against other diseases with copper accumulation [16]. In our routine practice we do Timm’s stain for every liver biopsy and hepatectomy. Next to evaluating copper accumulation for diagnosis WD disease, it can help to define late stage fibrosis [18]. It should be keep in mind, copper accumulation can be seen in other diseases such as cholestatic liver diseases, alcoholic liver disease and idiopathic copper toxicosis [6]. In chronic cholestasis and non WD cirrhosis, copper staining is usually limited to periseptal areas with a patchy/focal distribution (Figure 6). It is suggested that that in the absence of advanced fibrosis (or WD), a positive rhodanine stain for copper argues strongly in favor of chronic biliary diseases and against other liver diseases [19]. Of note, marked hepatic copper overload mimicking WD has been described in children with MDR3 deficiency [8]. It is important to remember that negative staining for both copper and copper-associated protein does not exclude the diagnosis of WD.
Different sensitivities of copper stains in the same case (A. Timm, B. Rhodanine, C. Orcein).
Periseptal copper accumulation in a non WD cirrhosis (Timm).
In equivocal cases, measurement of liver copper content is recommended as the next step for diagnosis of WD. A 5-fold increase of hepatic copper concentration is considered as diagnostic for diagnosis of hepatic WD [5]. In a more strict definition, a copper content >250 μg/g dry weight (normal value <50 mg/g dry weight) in adult patients without cholestasis is accepted as diagnostic for WD. Probably depending on sampling error due to nonhomogeneous copper distribution in the liver, lower values are reported in up to 20% of patients with WD. The exactness of liver copper estimation is moved forward with an optimal measured biopsy sample (ideally >1 cm long, min. 0.5 cm) that ought to be put on a little piece of paper for drying, and in a dry plastic copper-free holder for atomic absorption analysis on fresh tissue [13]. Hepatic copper levels in advanced stage chronic biliary diseases in adults and children often exceed 250 mg/100 g of dried liver, sometimes reaching levels higher than those observed in WD [19]. In spite of the fact that utilize of dried tissue has been proposed for tissue copper quantitation, the utilize of formalin-fixed, paraffin-embedded (FFPE) tissue is said fair as valuable. Utilizing FFPE tissue specimens evacuates the specialized troubles related to dried unfixed tissue, as well as gives the same tissue for histopathological and quantitative assessments. Since copper accumulation may well be non homogenious indeed even in most progressed cases of WD, the availability of light microscopy on the same tissue being evaluated for copper may well be a really valuable tool in mostly tending to this potential examining inclination tissue quantitation of copper is subject to [17]. Although liver copper content is a useful parameter, but a value below 250 μg/g does not exclude WD. Diagnosis requires the combination of a variety of clinical and biochemical tests [5].
WD is a curable disease, but early diagnosis is essential to stop the progression to cirrhosis or worsening of the neurological and psychiatric conditions. As a treatable disease, WD should be detected by any health professionals at any care level. If WD is not recognized and adequately treated, the progression of liver disease to cirrhosis and liver failure can be rapid or irreversible brain damage can occur. Unfortunately, even though of all advances, the diagnosis of WD shows up frequently compelling, due to the variability of its clinical manifestation and to the complexity of the microscopic findings within the liver biopsy. Liver histopathology, in reality, does not show a unique morphology, but it may appear in different patterns. From a pathologist’s perspective, when evaluating the liver biopsies, WD should be included in the differential diagnosis especially in pediatric age and also cryptogenic adult cases.
The author would like to thank to Dr. Emine Samdanci as a team member of hepatopathology and to Dr. Seyma Eren and Dr. Meryem Uludag for photography.
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