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Colorectal cancer is the third most common cancer worldwide, and in many parts of the western world, it is the second leading cause of cancer-related deaths. This book covers colon cancer metastasis from the most fundamental aspects to clinical practice. Major topics include physiopathology, genetic and epigenetic controls, cancer initiating cells, epithelial-mesenchymal transition, growth factors and signalling, cell adhesion, natures of liver metastasis, angiogenesis and lymphangiogenesis, inflammatory response, prognostic markers, sentinel node and staging, and finally diagnosis and treatment. Each chapter has been contributed by leaders in the field. A key feature is that it connects with a large readership including students, fundamentalists and clinicians. Another specific feature of the book is that the chapters are written in a didactic and illustrative fashion. These characteristics coupled with the choice of the topics and authors, makes this book a reference in the field. It represents an essential acquisition for medical libraries, clinicians as well as medical and graduate students.

Molecular biology --- Immunology. Immunopathology --- Oncology. Neoplasms --- Pathology --- monoklonale antilichamen --- polyklonale antilichamen --- immunologie --- pathologie --- oncologie --- moleculaire biologie

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It is an honor and a pleasure to present the proceedings of the 24th European Conference on Object-Oriented Programming (ECOOP 2010) held in Maribor, Slovenia. As always, the conference catered to a broad audience of members of industry and academia and consequently o?ered a mix of results in both applied andtheoreticalresearchinobjectorientation. Inall,24technicalpapers,12wo- shops,6tutorials,adoctoralsymposiumandaspecialsessiononempiricalme- ods were organized. Additionally, a seriesof seven summer school sessionsran in parallel with the technical paper sessions. A junior and a senior Dahl-Nygaard award were selected by AITO leading to two keynote talks, complemented by an invited keynote on the subject of empirical methods as applied to software practice. The banquet speech was given by Oscar Nierstrasz. The ECOOP P- gram Committee also selected a best paper award. These proceedings consist of 24 papers and one extended abstract selected from 108 submissions. Each paper received3 and as many as 6 reviewsfrom a ProgramCommittee consisting of 30 internationally reputed researchers. A long and intensive virtual discussion via CyberChairPROled to an author response period and to another week of online deliberation. This wasroundedo?by a ProgramCommittee meeting late Feb- aryinBrussels. Itwasagainshownthatnothingmatchesaphysicalmeeting,and that it in several instances is even indispensable, in spite of the huge e?ort that is required to organize it. The Program Committee discussions followed Oscar Nierstrasz' Champion pattern: for a paper to be accepted at least one manifest proponent is required. Program Committee members were allowed to submit a paper, but these were subjected to a higher level of scrutiny.

Logic --- Immunology. Immunopathology --- Computer science --- Programming --- Computer architecture. Operating systems --- Computer. Automation --- monoklonale antilichamen --- polyklonale antilichamen --- immunologie --- computers --- computerbesturingssystemen --- programmeren (informatica) --- programmeertalen --- software engineering --- computernetwerken

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Biological techniques --- Histology. Cytology --- Immunology. Immunopathology --- Semiology. Diagnosis. Symptomatology --- Human medicine --- Biotechnology --- monoklonale antilichamen --- polyklonale antilichamen --- immunologie --- klinische laboratoria --- biologische technieken --- medische laboratoriumtechnologie --- biotechnologie --- cytologie --- histologie

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Combiningtwodifferentscienti?cdisciplines-morphologyandimmunochemistry- immunohistochemistryhasdevelopedasanimportantinstrumentinresearchand clinicalpathology. A basicunderstandingofunderlying principlesandpotential problemsisunavoidableifyouwanttobesuccessfulinyouruseofimmunohis- chemistry,aswellasingettingyourpaperspublishedandyourresearchgrants funded. Whilemanyexcellenttextsandmonographsexistwhichcovervariousaspects ofimmunohistochemistry,thelackofaconcisecomprehensiveguidetousing thesemethodswasamajormotivationforwritingthisbook. Ourintentionwasto createaneasy-to-readandfocusedresourcebasedonstate-of-the-artinformation forabroadaudiencerangingfromstudentsandtechnicalassistantstoexperienced researchers. Thishandbookhasaconciseformat,withprotocolsandinstructionsfor methodsimmediatelyfollowingtheshortintroductorytheoreticalmaterialineach chapter. BeingconsciousofthegrowingroleofInternetasaninformationsource, wehavefounditreasonableinmanycasestosubstitutecitingbooksandjournal publications with corresponding Internet websites. Where possible, commercial sourcesofreagents,kits,andequipmentarelistedthroughoutthetextinsteadof inaseparateindex. Thougheachchapterissmallandintroductory,thishandbook itself is self-suf?cient and provides a comprehensive look at the principles of immunohistochemistry. For readers wanting further depth of knowledge, each chapterisbackedupbyashortlistofcarefullyselectedoriginalarticles. Duringthelastdecade,pioneeringeffortsofhistochemistshaveledtoan- menseimprovementinthereagentsandprotocols. Theresearcherisurgedalwaysto determinethereasonforeverymethodandstepbeforedoingit. Thishandbookis intendedtohelpreaderstoavoidtroublesinthechoiceofanadequatemethod,which happenswhenusingstandardtextbooks. Forthishandbook,wecarefullyselected establishedmethodsandeasy-to-adoptprotocols,payingattentiontomoderndev- opmentsinimmunohistochemistry,suchasantigenretrieval,signalampli?cation, the use of epitope tags in immunohistochemistry, multiple immunolabeling or diagnosticimmunohistochemistry. Eachofthemethodsdescribedinthishandbook v vi Preface was provedby the authors; many of these methods are routinely used in daily practiceintheirinstitute. Allthepracticalmethodsadvocatedareclearlydescribed, withaccompanyingtables,andtheresultsobtainableareillustratedwithcolour micrographs. Acknowledgements We thank Vera Samoilova forthe perfect technical assistance and other colleaguesfromtheMu¨nsterUniversityClinicforsharingprobesandreagents. IgorB. BuchwalowandWernerBo¨cker Mu¨nster Contents 1 AntibodiesforImmunohistochemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 1. 1 StructureofAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1. 2 PolyclonalAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1. 3 MouseMonoclonalAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1. 4 RabbitMonoclonalAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1. 5 ProteinAandProteinGinImmunohistochemistry . . . . . . . . . . . . . . . . . . . 7 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 2 AntibodyLabelingandtheChoiceoftheLabel . . . . . . . . . . . . . . . . . . . . . . . . 9 2. 1 CovalentLabelingofAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 2. 2 Non-CovalentLabelingofPrimaryAntibodieswith LabeledFabFragments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10 2. 3 EnzymeLabelsforLightMicroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 2. 4 FluorophoreLabelsforFluorescenceMicroscopy . . . . . . . . . . . . . . . . . . . 15 2. 5 ColloidalGoldLabelsforElectronMicroscopy . . . . . . . . . . . . . . . . . . . . . 16 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 3 ProbesProcessinginImmunohistochemistry . . . . . . . . . . . . . . . . . . . . . . . . . . 19 3. 1 FixationinImmunohistochemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 3. 1. 1 FixationinAlcoholsandAcetone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 3. 1. 2 FixationinFormaldehyde . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 3. 1. 3 EffectofFormaldehydeFixationon Antigen-AntibodyBinding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 3. 2 Paraf?nSectionsforImmunohistochemicalAnalysis . . . . . . . . . . . . . . . 22 3. 2. 1 EmbeddingandCutting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 3. 2. 2 MountingParaf?nSectionsontoSlides . . . . . . . . . . . . . . . . . . . . . . . 23 3. 3 CryosectionsforImmunohistochemicalAnalysis . . . . . . . . . . . . . . . . . . . . 24 3. 4 BuffersforWashingandAntibodyDilution . . . . . . . . . . . . . . . . . . . . . . . . . 25 3. 5 MountingFollowingImmunohistochemicalStaining . . . . . . . . . . . . . . . . 27 3. 6 StorageFollowingImmunohistochemicalStaining . . . . . . . . . . . . . . . . . . 28 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 vii viii Contents 4 WorkingwithAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 4. 1 DirectImmunostainingMethod . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 4. 2 IndirectImmunostainingMethod . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 4. 3 TheChoiceofAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 4. 3. 1 TheChoiceofPrimaryAntibodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35 4. 3. 2 TheChoiceofSecondaryAntibodies . . . . . . . . . . . . . . . . . . . . . . . . .

Biological techniques --- Histology. Cytology --- Immunology. Immunopathology --- Semiology. Diagnosis. Symptomatology --- Human medicine --- Biotechnology --- monoklonale antilichamen --- polyklonale antilichamen --- immunologie --- klinische laboratoria --- biologische technieken --- medische laboratoriumtechnologie --- biotechnologie --- cytologie --- histologie --- Immuncytochemie. --- Immunohistochemistry --- Immunohistochemistry.

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The fourth edition of The Immunoassay Handbook provides an excellent, thoroughly updated guide to the science, technology and applications of ELISA and other immunoassays, including a wealth of practical advice. It encompasses a wide range of methods and gives an insight into the latest developments and applications in clinical and veterinary practice and in pharmaceutical and life science research. Highly illustrated and clearly written, this award-winning reference work provides an excellent guide to this fast-growing field. Revised and extensively updated, with over 30% new

General biochemistry --- Immunology. Immunopathology --- monoklonale antilichamen --- polyklonale antilichamen --- klinische chemie --- laboratoriumdiagnostiek --- laboratoriumpraxis --- medische laboratoriumtechnologie --- proteomics --- biochemie --- zwangerschap --- immunochemie --- Immunoassay --- methods --- standards --- 57.083.3 --- 57.083.3 Immunological methods and techniques --- Immunological methods and techniques --- Antigens --- Immunoglobulins --- Immunology --- Immunodiagnosis --- Immunoassay - Handbooks, manuals, etc. --- Immunoassay - methods --- Immunoassay - standards --- methods. --- standards.