{"product_id":"the-agt-cytogenetics-laboratory-manual-isbn-9781119061229","title":"The AGT Cytogenetics Laboratory Manual","description":"\u003cp\u003eCytogenetics is the study of chromosome morphology, structure, pathology, function, and behavior. The field has evolved to embrace molecular cytogenetic changes, now termed cytogenomics. \u003cbr\u003eCytogeneticists utilize an assortment of procedures to investigate the full complement of chromosomes and\/or a targeted region within a specific chromosome in metaphase or interphase.  Tools include routine analysis of G-banded chromosomes, specialized stains that address specific chromosomal structures, and molecular probes, such as fluorescence in situ hybridization (FISH) and chromosome microarray analysis, which employ a variety of methods to highlight a region as small as a single, specific genetic sequence under investigation.\u003c\/p\u003e \u003cp\u003e\u003ci\u003eThe AGT Cytogenetics Laboratory Manual, Fourth Edition\u003c\/i\u003e offers a comprehensive description of the diagnostic tests offered by the clinical laboratory and explains the science behind them. One of the most valuable assets is its rich compilation of laboratory-tested protocols currently being used in leading laboratories, along with practical advice for nearly every area of interest to cytogeneticists. In addition to covering essential topics that have been the backbone of cytogenetics for over 60 years, such as the basic components of a cell, use of a microscope, human tissue processing for cytogenetic analysis (prenatal, constitutional, and neoplastic), laboratory safety, and the mechanisms behind chromosome rearrangement and aneuploidy, this edition introduces new and expanded chapters by experts in the field.  Some of these new topics include a unique collection of chromosome heteromorphisms; clinical examples of genomic imprinting; an example-driven overview of chromosomal microarray; mathematics specifically geared for the cytogeneticist; usage of ISCN’s cytogenetic language to describe chromosome changes; tips for laboratory management; examples of laboratory information systems; a collection of internet and library resources; and a special chapter on animal chromosomes for the research and zoo cytogeneticist. The range of topics is thus broad yet comprehensive, offering the student a resource that teaches the procedures performed in the cytogenetics laboratory environment, and the laboratory professional with a peer-reviewed reference that explores the basis of each of these procedures. This makes it a useful resource for researchers, clinicians, and lab professionals, as well as students in a university or medical school setting.\u003c\/p\u003eDie Zyto- oder Zellgenetik untersucht die Morphologie, Struktur, Pathologie, Funktion und das Verhalten von Chromosomen. Entstanden ist dieses Teilgebiet der Genetik, um zytogenetische Veränderungen auf Molekularebene abzubilden. Heute spricht man in diesem Zusammenhang von der Zellgenomik.\u003cbr\u003e Zellgenetiker greifen auf eine ganze Reihe von Verfahren zurück, um Chromosomen und\/oder eine Zielregion eines bestimmten Chromosom in der Meta- oder Interphase zu untersuchen. Zu den Tools gehören Routineanalysen von Chromosomen (G-Banding), spezielle Stains für bestimmte Chromosomenstrukturen und molekulare Sonden, z. B. im Bereich der Fluoreszenz-in-situ-Hybridisierung (FISH) sowie der Chromosomenanalyse auf Microarray-Basis. Zum Einsatz kommen eine Vielzahl von Methoden, um eine zu untersuchenden Region hervorzuheben, der so klein ist, wie eine einzige, spezifische Gensequenz.\u003cbr\u003e \u003cp\u003eContributing authors xxvii\u003c\/p\u003e \u003cp\u003ePreface xxix\u003c\/p\u003e \u003cp\u003eAcknowledgments xxxi\u003c\/p\u003e \u003cp\u003e\u003cb\u003e1 The cell and cell division 1\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eMargaret J. Barch and Helen J. Lawce\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e1.1 The cell 1\u003c\/p\u003e \u003cp\u003e1.2 The cell cycle 14\u003c\/p\u003e \u003cp\u003e1.3 Recombinant DNA techniques 19\u003c\/p\u003e \u003cp\u003e1.4 The human genome 21\u003c\/p\u003e \u003cp\u003eReferences 22\u003c\/p\u003e \u003cp\u003e\u003cb\u003e2 Cytogenetics: an overview 25\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eHelen J. Lawce and Michael G. Brown\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e2.1 Introduction 25\u003c\/p\u003e \u003cp\u003e2.2 History of human cytogenetics 25\u003c\/p\u003e \u003cp\u003e2.3 Cytogenetics methods 29\u003c\/p\u003e \u003cp\u003e2.4 Slide‐making 49\u003c\/p\u003e \u003cp\u003e2.5 Chromosome staining 58\u003c\/p\u003e \u003cp\u003e2.6 Chromosome microscopy\/analysis 59\u003c\/p\u003e \u003cp\u003e2.7 Laboratory procedure manual 69\u003c\/p\u003e \u003cp\u003eReferences 70\u003c\/p\u003e \u003cp\u003eContributed protocols 75\u003c\/p\u003e \u003cp\u003eProtocol 2.1 Slide‐making 75\u003c\/p\u003e \u003cp\u003eProtocol 2.2 Slide‐making 76\u003c\/p\u003e \u003cp\u003eProtocol 2.3 Making wet slides for chromosome analysis 78\u003c\/p\u003e \u003cp\u003eProtocol 2.4 Slide‐making 82\u003c\/p\u003e \u003cp\u003eProtocol 2.5 Slide preparation 82\u003c\/p\u003e \u003cp\u003eProtocol 2.6 Slide preparation procedure 84\u003c\/p\u003e \u003cp\u003e\u003cb\u003e3 Peripheral blood cytogenetic methods 87\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eHelen J. Lawce and Michael G. Brown\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e3.1 Using peripheral blood for cytogenetic analysis 87\u003c\/p\u003e \u003cp\u003e3.2 Special uses of peripheral blood cultures 88\u003c\/p\u003e \u003cp\u003e3.3 Peripheral blood constituents 89\u003c\/p\u003e \u003cp\u003e3.4 Specimen handling 91\u003c\/p\u003e \u003cp\u003e3.5 Cell culture equipment and supplies 93\u003c\/p\u003e \u003cp\u003e3.6 Harvesting peripheral blood cultures 95\u003c\/p\u003e \u003cp\u003e3.7 Chromosome analysis of peripheral blood 95\u003c\/p\u003e \u003cp\u003e3.8 Storage of fixed specimens 95\u003c\/p\u003e \u003cp\u003eAcknowledgments 95\u003c\/p\u003e \u003cp\u003eReferences 95\u003c\/p\u003e \u003cp\u003eContributed protocols 98\u003c\/p\u003e \u003cp\u003eProtocol 3.1 Blood culture and harvest procedure 98\u003c\/p\u003e \u003cp\u003eProtocol 3.2 High‐resolution peripheral blood method 100\u003c\/p\u003e \u003cp\u003eProtocol 3.3 Constitutional cytogenetic studies on peripheral blood 108\u003c\/p\u003e \u003cp\u003eProtocol 3.4 Blood culture and harvest procedure for microarray confirmation studies 115\u003c\/p\u003e \u003cp\u003e\u003cb\u003e4 General cell culture principles and fibroblast culture 119\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eDebra F. Saxe, Kristin M. May and Jean H. Priest\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e4.1 Definitions of a culture 119\u003c\/p\u003e \u003cp\u003e4.2 Basic considerations in cell culture 121\u003c\/p\u003e \u003cp\u003e4.3 Fibroblast culture 128\u003c\/p\u003e \u003cp\u003e4.4 Lymphoblastoid cell lines 132\u003c\/p\u003e \u003cp\u003eGlossary 132\u003c\/p\u003e \u003cp\u003eReference 133\u003c\/p\u003e \u003cp\u003eAdditional readings 133\u003c\/p\u003e \u003cp\u003eContributed protocols section 134\u003c\/p\u003e \u003cp\u003eProtocol 4.1 Solid tissue collection for establishing cultures 134\u003c\/p\u003e \u003cp\u003eProtocol 4.2 Solid tissue transport and sendout media 135\u003c\/p\u003e \u003cp\u003eProtocol 4.3 Tissue culture reagents 138\u003c\/p\u003e \u003cp\u003eProtocol 4.4 Phosphate buffer solution deficient in Ca2+ and Mg2+ 141\u003c\/p\u003e \u003cp\u003eProtocol 4.5 Solid tissue and fibroblast culture setup 141\u003c\/p\u003e \u003cp\u003eProtocol 4.6 Solid tissue setup and processing 142\u003c\/p\u003e \u003cp\u003eProtocol 4.7 Flask and coverslip setup for POC\/fibroblast cultures 145\u003c\/p\u003e \u003cp\u003eProtocol 4.8 Coverslip setup for solid tissue biopsy specimens 147\u003c\/p\u003e \u003cp\u003eProtocol 4.9 Solid tissue (fibroblast) culturing and harvesting 150\u003c\/p\u003e \u003cp\u003eProtocol 4.10 Fibroblast culture maintenance: media feeding and changing 154\u003c\/p\u003e \u003cp\u003eProtocol 4.11 Routine subculture of fibroblast cultures 155\u003c\/p\u003e \u003cp\u003eProtocol 4.12 Manual harvest for flasks 157\u003c\/p\u003e \u003cp\u003eProtocol 4.13 Treated media for contamination 158\u003c\/p\u003e \u003cp\u003eProtocol 4.14 Fungizone–mycostatin solution for treatment of fungus\/yeast contaminated cultures 158\u003c\/p\u003e \u003cp\u003eProtocol 4.15 Mycoplasma testing 159\u003c\/p\u003e \u003cp\u003eProtocol 4.16 Plating efficiency of serum 160\u003c\/p\u003e \u003cp\u003eProtocol 4.17 Routine replication plating for human diploid cells 160\u003c\/p\u003e \u003cp\u003eProtocol 4.18 Cell counting chamber method 161\u003c\/p\u003e \u003cp\u003eProtocol 4.19 Cell viability by dye exclusion 161\u003c\/p\u003e \u003cp\u003eProtocol 4.20 Mitotic index 161\u003c\/p\u003e \u003cp\u003eProtocol 4.21 Growth rate‐estimation of mean population doubling time during logarithmic growth 162\u003c\/p\u003e \u003cp\u003eProtocol 4.22 Maintenance of fibroblast cultures as non‐mitotic population 163\u003c\/p\u003e \u003cp\u003eProtocol 4.23 Synchronization at S‐phase with BrdU 163\u003c\/p\u003e \u003cp\u003eProtocol 4.24 Making direct FISH preparations from abortus tissue 164\u003c\/p\u003e \u003cp\u003eProtocol 4.25 Cryopreservation 165\u003c\/p\u003e \u003cp\u003eProtocol 4.26 Cryopreservation with Nalgene cryogenic container 166\u003c\/p\u003e \u003cp\u003eProtocol 4.27 Lymphoblastoid lines 167\u003c\/p\u003e \u003cp\u003eProtocol 4.28 Freezing tissue cultures (cryopreservation) 171\u003c\/p\u003e \u003cp\u003e\u003cb\u003e5 Prenatal chromosome diagnosis 173\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eKristin M. May, Debra F. Saxe and Jean H. Priest\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e5.1 Introduction 173\u003c\/p\u003e \u003cp\u003e5.2 Amniotic fluid 173\u003c\/p\u003e \u003cp\u003e5.3 Culture of amniotic fluid 175\u003c\/p\u003e \u003cp\u003e5.4 Analysis of amniotic fluid 178\u003c\/p\u003e \u003cp\u003e5.5 Chorionic villus sampling 180\u003c\/p\u003e \u003cp\u003e5.6 Analysis of chorionic villi 184\u003c\/p\u003e \u003cp\u003eReferences 186\u003c\/p\u003e \u003cp\u003eContributed protocols section 188\u003c\/p\u003e \u003cp\u003eProtocol 5.1 Amniotic fluid culture setup and routine maintenance 188\u003c\/p\u003e \u003cp\u003eProtocol 5.2 Coverslip (in situ) harvest procedure for chromosome preparations from amniotic fluid, CVS, or tissues (manual method) 191\u003c\/p\u003e \u003cp\u003eProtocol 5.3 Harvest of flask amniocyte cultures 193\u003c\/p\u003e \u003cp\u003eProtocol 5.4 Amniotic fluid culturing, subculturing, and harvesting (flask method) 195\u003c\/p\u003e \u003cp\u003eProtocol 5.5 Criteria for interpreting mosaic amniotic fluid cultures 198\u003c\/p\u003e \u003cp\u003eProtocol 5.6 Chorionic villi sampling – setup, direct harvest, and culture 199\u003c\/p\u003e \u003cp\u003eProtocol 5.7 Chorionic villus sampling 204\u003c\/p\u003e \u003cp\u003eProtocol 5.8 G‐Banding with Leishman’s stain (GTL) 208\u003c\/p\u003e \u003cp\u003eProtocol 5.9 Cystic hygroma fluid protocol 209\u003c\/p\u003e \u003cp\u003e\u003cb\u003e6 Chromosome stains 213\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eHelen J. Lawce\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e6.1 Introduction 213\u003c\/p\u003e \u003cp\u003e6.2 Chromosome banding methods 220\u003c\/p\u003e \u003cp\u003e6.3 5-bromo-2′-deoxyuridine methodologies 246\u003c\/p\u003e \u003cp\u003e6.4 T‐banding\/CT‐banding 252\u003c\/p\u003e \u003cp\u003e6.5 Antibody banding and restriction endonuclease banding 252\u003c\/p\u003e \u003cp\u003e6.6 Destaining slides 252\u003c\/p\u003e \u003cp\u003e6.7 FISH DAPI bands 252\u003c\/p\u003e \u003cp\u003e6.8 Sequential staining 253\u003c\/p\u003e \u003cp\u003eAcknowledgments 253\u003c\/p\u003e \u003cp\u003eReferences 253\u003c\/p\u003e \u003cp\u003eContributed protocols section 266\u003c\/p\u003e \u003cp\u003eProtocol 6.1 Conventional Giemsa staining (unbanded) 266\u003c\/p\u003e \u003cp\u003eProtocol 6.2 Leishman’s stain 266\u003c\/p\u003e \u003cp\u003eProtocol 6.3 Quinacrine mustard chromosome staining (Q‐bands) 266\u003c\/p\u003e \u003cp\u003eProtocol 6.4 C‐banding 268\u003c\/p\u003e \u003cp\u003eProtocol 6.5 C‐banding 270\u003c\/p\u003e \u003cp\u003eProtocol 6.6 C‐banding 271\u003c\/p\u003e \u003cp\u003eProtocol 6.7 C‐banding of blood slides 272\u003c\/p\u003e \u003cp\u003eProtocol 6.8 Giemsa‐11 staining technique 274\u003c\/p\u003e \u003cp\u003eProtocol 6.9 Distamycin A\/DAPI staining 275\u003c\/p\u003e \u003cp\u003eProtocol 6.10 Chromomycin\/methyl green and chromomycin\/distamycin fluorescent R‐banding method 277\u003c\/p\u003e \u003cp\u003eProtocol 6.11 Bone marrow and cancer blood G‐banding 278\u003c\/p\u003e \u003cp\u003eProtocol 6.12 Trypsin G‐banding 280\u003c\/p\u003e \u003cp\u003eProtocol 6.13 Giemsa‐trypsin banding with Wright stain (GTW) for suspension culture slides and in situ culture coverslips 281\u003c\/p\u003e \u003cp\u003eProtocol 6.14 G‐banding blood lymphocyte slides 284\u003c\/p\u003e \u003cp\u003eProtocol 6.15 Cd staining 285\u003c\/p\u003e \u003cp\u003eProtocol 6.16 CREST\/CENP antibody staining 286\u003c\/p\u003e \u003cp\u003eProtocol 6.17 AgNOR (silver staining) 287\u003c\/p\u003e \u003cp\u003eProtocol 6.18 Sister chromatid exchange blood culture and staining 289\u003c\/p\u003e \u003cp\u003eProtocol 6.19 Sister chromatid exchange fibroblast culture and staining 291\u003c\/p\u003e \u003cp\u003eProtocol 6.20 T‐banding by thermal denaturation 294\u003c\/p\u003e \u003cp\u003eProtocol 6.21 CT‐banding 295\u003c\/p\u003e \u003cp\u003eProtocol 6.22 Lymphocyte culture and staining procedures for late replication analysis 295\u003c\/p\u003e \u003cp\u003eProtocol 6.23 Destaining and sequential staining of slides 298\u003c\/p\u003e \u003cp\u003eProtocol 6.24 Restaining permanently mounted slides 299\u003c\/p\u003e \u003cp\u003e\u003cb\u003e7 Human chromosomes: identification and variations 301\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eHelen J. Lawce and Luke Boyd\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e7.1 Understanding the basics 301\u003c\/p\u003e \u003cp\u003e7.2 Description of human chromosome shapes 302\u003c\/p\u003e \u003cp\u003e7.3 Determination of G‐banded chromosome resolution 355\u003c\/p\u003e \u003cp\u003e\u003cb\u003e8 ISCN: the universal language of cytogenetics 359\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eMarilyn S. Arsham and Lisa G. Shaffer\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e8.1 Introduction 359\u003c\/p\u003e \u003cp\u003e8.2 Language 359\u003c\/p\u003e \u003cp\u003e8.3 Karyotype 364\u003c\/p\u003e \u003cp\u003e8.4 Numerical events 378\u003c\/p\u003e \u003cp\u003e8.5 Structural events 380\u003c\/p\u003e \u003cp\u003e8.6 Derivative chromosomes (der) 394\u003c\/p\u003e \u003cp\u003e8.7 Symbols of uncertainty 397\u003c\/p\u003e \u003cp\u003e8.8 Random versus reportable 403\u003c\/p\u003e \u003cp\u003e8.9 Multiple cell lines and clones\u003c\/p\u003e \u003cp\u003e8.10 Fluorescence in situ hybridization 408\u003c\/p\u003e \u003cp\u003e8.11 Microarray (arr) and region‐specific assay (rsa) 420\u003c\/p\u003e \u003cp\u003e8.12 Conclusion 422\u003c\/p\u003e \u003cp\u003eAcknowledgments 422\u003c\/p\u003e \u003cp\u003eAddendum for ISCN 2016 updates 426\u003c\/p\u003e \u003cp\u003eReferences 426\u003c\/p\u003e \u003cp\u003e\u003cb\u003e9 Constitutional chromosome abnormalities 429\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eKathleen Kaiser\u003c\/i\u003e\u003ci\u003e‐Rogers\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e9.1 Numerical abnormalities 429\u003c\/p\u003e \u003cp\u003e9.2 Structural rearrangements 444\u003c\/p\u003e \u003cp\u003eReferences 472\u003c\/p\u003e \u003cp\u003e\u003cb\u003e10 Genomic imprinting 481\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eR. Ellen Magenis\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e10.1 Introduction 481\u003c\/p\u003e \u003cp\u003e10.2 Human genomic disease and imprinting 488\u003c\/p\u003e \u003cp\u003e10.3 Germ cell tumors – UPD and imprinting 493\u003c\/p\u003e \u003cp\u003eGlossary 494\u003c\/p\u003e \u003cp\u003eReferences 496\u003c\/p\u003e \u003cp\u003e\u003cb\u003e11 Cytogenetic analysis of hematologic malignant diseases 499\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eNyla A. Heerema\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e11.1 Introduction 499\u003c\/p\u003e \u003cp\u003e11.2 Myeloid leukemias 508\u003c\/p\u003e \u003cp\u003e11.3 Myelodysplastic syndromes 514\u003c\/p\u003e \u003cp\u003e11.4 Myeloproliferative neoplasms 515\u003c\/p\u003e \u003cp\u003e11.5 B‐ and T‐cell lymphoid neoplasms 517\u003c\/p\u003e \u003cp\u003e11.6 Lymphomas 522\u003c\/p\u003e \u003cp\u003e11.7 Laboratory practices 525\u003c\/p\u003e \u003cp\u003eAcknowledgments 533\u003c\/p\u003e \u003cp\u003eGlossary of hematopoietic malignancies 533\u003c\/p\u003e \u003cp\u003eReferences 535\u003c\/p\u003e \u003cp\u003eContributed protocols section 553\u003c\/p\u003e \u003cp\u003eProtocol 11.1 Cancer cytogenetics procedure 553\u003c\/p\u003e \u003cp\u003eProtocol 11.2 Bone marrow\/leukemic peripheral blood setup and harvest procedure 558\u003c\/p\u003e \u003cp\u003eProtocol 11.3 Bone marrow and leukemic blood culture and harvest procedure using DSP30 CPG oligonucleotide\/interleukin‐2 for B‐cell mitogenic stimulation 560\u003c\/p\u003e \u003cp\u003eProtocol 11.4 Culture of CpG‐stimulated peripheral blood and bone marrow in chroniclymphocytic leukemia 562\u003c\/p\u003e \u003cp\u003eProtocol 11.5 Plasma cell separation and harvest procedure for FISH analysis 567\u003c\/p\u003e \u003cp\u003eProtocol 11.6 Plasma cell separation and harvest procedure for FISH 569\u003c\/p\u003e \u003cp\u003eProtocol 11.7 Bone marrow GTG‐banding 571\u003c\/p\u003e \u003cp\u003eProtocol 11.8 GTW banding procedure (G‐bands by trypsin using Wright stain) 573\u003c\/p\u003e \u003cp\u003e\u003cb\u003e12 Cytogenetic methods and findings in human solid tumors 577\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eMarilu Nelson\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e12.1 Introduction 577\u003c\/p\u003e \u003cp\u003e12.2 Processing tumor specimens 579\u003c\/p\u003e \u003cp\u003e12.3 Recurrent cytogenetic abnormalities 592\u003c\/p\u003e \u003cp\u003e12.4 Molecular genetic and cytogenetic techniques 608\u003c\/p\u003e \u003cp\u003e12.5 Conclusion 612\u003c\/p\u003e \u003cp\u003eGlossary 612\u003c\/p\u003e \u003cp\u003eReferences 613\u003c\/p\u003e \u003cp\u003eContributed protocol section 631\u003c\/p\u003e \u003cp\u003eProtocol 12.1 Solid tumor cell culture and harvest 631\u003c\/p\u003e \u003cp\u003eProtocol 12.2 Solid tumor cell culture and harvest 637\u003c\/p\u003e \u003cp\u003eProtocol 12.3 Solid tumor culture 643\u003c\/p\u003e \u003cp\u003eProtocol 12.4 Solid tumor harvest: monolayer and flask methods 644\u003c\/p\u003e \u003cp\u003eProtocol 12.5 Solid tumor culturing and harvesting 646\u003c\/p\u003e \u003cp\u003e\u003cb\u003e13 Chromosome instability syndromes 653\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eYassmine Akkari\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e13.1 Introduction 653\u003c\/p\u003e \u003cp\u003e13.2 Fanconi anemia 656\u003c\/p\u003e \u003cp\u003e13.3 Bloom syndrome 658\u003c\/p\u003e \u003cp\u003e13.4 Ataxia–telangiectasia 658\u003c\/p\u003e \u003cp\u003e13.5 Nijmegen breakage syndrome 659\u003c\/p\u003e \u003cp\u003e13.6 Immunodeficiency, centromeric instability, and facial anomalies syndrome 660\u003c\/p\u003e \u003cp\u003e13.7 Roberts syndrome 661\u003c\/p\u003e \u003cp\u003e13.8 Werner syndrome 661\u003c\/p\u003e \u003cp\u003e13.9 Rothmund–Thomson syndrome 662\u003c\/p\u003e \u003cp\u003e13.10 Proficiency testing 662\u003c\/p\u003e \u003cp\u003eGlossary 662\u003c\/p\u003e \u003cp\u003eReferences 667\u003c\/p\u003e \u003cp\u003eContributed protocol section 671\u003c\/p\u003e \u003cp\u003eProtocol 13.1 Fanconi anemia chromosome breakage procedure for whole blood 671\u003c\/p\u003e \u003cp\u003eProtocol 13.2 Supplemental procedure; Ficoll separation of whole blood 675\u003c\/p\u003e \u003cp\u003eProtocol 13.3 Fanconi anemia fibroblast set up, culture, subculture, and harvest procedure 676\u003c\/p\u003e \u003cp\u003eProtocol 13.4 Fanconi anemia chromosome breakage analysis policy 681\u003c\/p\u003e \u003cp\u003eProtocol 13.5 Table for breakage studies result interpretation 682\u003c\/p\u003e \u003cp\u003eProtocol 13.6 Fanconi anemia 684\u003c\/p\u003e \u003cp\u003e\u003cb\u003e14 Microscopy and imaging 687\u003cbr\u003e\u003c\/b\u003eMargaret J. Barch and Helen J. Lawce\u003c\/p\u003e \u003cp\u003e14.1 The standard microscope 687\u003c\/p\u003e \u003cp\u003e14.2 Brightfield microscopy 695\u003c\/p\u003e \u003cp\u003e14.3 Fluorescence microscopy 697\u003c\/p\u003e \u003cp\u003e14.4 Specialized microscopy 699\u003c\/p\u003e \u003cp\u003e14.5 Capturing the microscopic image 701\u003c\/p\u003e \u003cp\u003eReferences 703\u003c\/p\u003e \u003cp\u003e\u003cb\u003e15 Computer imaging 705\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eChristine E. Haessig\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e15.1 Introduction 705\u003c\/p\u003e \u003cp\u003e15.2 Techniques to improve karyogram image quality 705\u003c\/p\u003e \u003cp\u003e15.3 Metaphase preparation 706\u003c\/p\u003e \u003cp\u003e15.4 Microscopy 706\u003c\/p\u003e \u003cp\u003e15.5 Image capture 707\u003c\/p\u003e \u003cp\u003e15.6 Enhancement 710\u003c\/p\u003e \u003cp\u003e15.7 Advanced contrast 710\u003c\/p\u003e \u003cp\u003e15.8 Macro programming 712\u003c\/p\u003e \u003cp\u003e15.9 FISH imaging 713\u003c\/p\u003e \u003cp\u003e15.10 Printing 715\u003c\/p\u003e \u003cp\u003e15.11 Quality control 715\u003c\/p\u003e \u003cp\u003e15.12 Archiving 715\u003c\/p\u003e \u003cp\u003eAcknowledgments 715\u003c\/p\u003e \u003cp\u003eReferences 715\u003c\/p\u003e \u003cp\u003e\u003cb\u003e16 Fluorescence in situ hybridization (FISH) 717\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eHelen J. Lawce and Jeffrey S. Sanford\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e16.1 Introduction 717\u003c\/p\u003e \u003cp\u003e16.2 Clinical applications of FISH probes 722\u003c\/p\u003e \u003cp\u003e16.3 Deletion\/duplication probes for constitutional abnormalities 730\u003c\/p\u003e \u003cp\u003e16.4 Hematology\/oncology and solid tumor probes 734\u003c\/p\u003e \u003cp\u003e16.5 Sources and characteristics of probes available to the clinical cytogenetics laboratory 736\u003c\/p\u003e \u003cp\u003e16.6 Special uses of probes 738\u003c\/p\u003e \u003cp\u003e16.7 Important FISH probe adjuvants 739\u003c\/p\u003e \u003cp\u003e16.8 Principles of FISH 741\u003c\/p\u003e \u003cp\u003e16.9 FISH methods – an overview 744\u003c\/p\u003e \u003cp\u003e16.10 FISH analysis and reporting 757\u003c\/p\u003e \u003cp\u003e16.11 FISH probe testing and validation 765\u003c\/p\u003e \u003cp\u003e16.12 FISH for special investigation 768\u003c\/p\u003e \u003cp\u003e16.13 Preimplantation genetic FISH 771\u003c\/p\u003e \u003cp\u003e16.14 Other applications 776\u003c\/p\u003e \u003cp\u003e16.15 Variants in FISH signal patterns 777\u003c\/p\u003e \u003cp\u003e16.16 Conclusion 777\u003c\/p\u003e \u003cp\u003eAcknowledgments 778\u003c\/p\u003e \u003cp\u003eGlossary 778\u003c\/p\u003e \u003cp\u003eReferences 780\u003c\/p\u003e \u003cp\u003eContributed protocols 790\u003c\/p\u003e \u003cp\u003eProtocol 16.1 FISH (fluorescence in situ hybridization) methods 790\u003c\/p\u003e \u003cp\u003eProtocol 16.2 LSI, CEP, and paint probe protocol 796\u003c\/p\u003e \u003cp\u003eProtocol 16.3 FISH protocol for multiprobe® FISH panels 799\u003c\/p\u003e \u003cp\u003eProtocol 16.4 Slide pretreatment with pepsin for FISH 800\u003c\/p\u003e \u003cp\u003eProtocol 16.5 Interphase FISH for amniotic fluid specimen aneuploidy 801\u003c\/p\u003e \u003cp\u003eProtocol 16.6 FISH on direct preparations from abortus tissue 803\u003c\/p\u003e \u003cp\u003eProtocol 16.7 FISH on cultured non‐mitotic abortus tissue 804\u003c\/p\u003e \u003cp\u003eProtocol 16.8 FISH on smears 806\u003c\/p\u003e \u003cp\u003eProtocol 16.9 FISH on very small samples 808\u003c\/p\u003e \u003cp\u003eProtocol 16.10 Paraffin‐embedded tissue FISH method 810\u003c\/p\u003e \u003cp\u003eProtocol 16.11 VP2000 automated slide processor method for FFPE FISH 811\u003c\/p\u003e \u003cp\u003eProtocol 16.12 Plasma cell targeted FISH 814\u003c\/p\u003e \u003cp\u003eProtocol 16.13 Plasma cell separation for interphase FISH using easy SEP magnet method 815\u003c\/p\u003e \u003cp\u003eProtocol 16.14 Preimplantation genetic testing (PGD) for aneuploidy 818\u003c\/p\u003e \u003cp\u003eProtocol 16.15 Preimplantation genetic testing (PGD) FISH for translocations 821\u003c\/p\u003e \u003cp\u003eProtocol 16.16 Post‐FISH BrdU antibody detection 823\u003c\/p\u003e \u003cp\u003eProtocol 16.17 Same‐day HER2 IQ‐FISH pharmDx™ for breast tissue 824\u003c\/p\u003e \u003cp\u003e\u003cb\u003e17 Multicolor FISH (SKY and M\u003c\/b\u003e\u003cb\u003e‐FISH) and CGH 833\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eTurid Knutsen\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e17.1 Introduction 833\u003c\/p\u003e \u003cp\u003e17.2 Multicolor FISH (SKY\/M‐FISH) 834\u003c\/p\u003e \u003cp\u003e17.3 Comparative genomic hybridization 849\u003c\/p\u003e \u003cp\u003e17.4 Conclusion 859\u003c\/p\u003e \u003cp\u003eAcknowledgments 859\u003c\/p\u003e \u003cp\u003eReferences 859\u003c\/p\u003e \u003cp\u003eContributed protocols section 864\u003c\/p\u003e \u003cp\u003eProtocol 17.1  Spectral karyotyping (SKY) 864\u003c\/p\u003e \u003cp\u003eProtocol 17.2  Spectral karyotyping (SKY) 877\u003c\/p\u003e \u003cp\u003eProtocol 17.3  DNA spectral karyotyping 878\u003c\/p\u003e \u003cp\u003eProtocol 17.4 Multicolor‐FISH method (M‐FISH) I 881\u003c\/p\u003e \u003cp\u003eProtocol 17.5 Multicolor FISH (M‐FISH) or 24‐color FISH II 884\u003c\/p\u003e \u003cp\u003eProtocol 17.6 Multicolor FISH (M‐FISH) III 888\u003c\/p\u003e \u003cp\u003eProtocol 17.7 Comparative genomic hybridization I 891\u003c\/p\u003e \u003cp\u003eProtocol 17.8 Comparative genomic hybridization II 898\u003c\/p\u003e \u003cp\u003e\u003cb\u003e18 Genomic microarray technologies for the cytogenetics laboratory 903\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eBhavana J. Davé and Warren G. Sanger\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e18.1 Introduction 903\u003c\/p\u003e \u003cp\u003e18.2 Applications 907\u003c\/p\u003e \u003cp\u003e18.3 Genomic microarray in a cytogenetics laboratory 913\u003c\/p\u003e \u003cp\u003e18.4 Conclusion 922\u003c\/p\u003e \u003cp\u003eAcknowledgment 922\u003c\/p\u003e \u003cp\u003eAuthors’ note 923\u003c\/p\u003e \u003cp\u003eReferences 923\u003c\/p\u003e \u003cp\u003e\u003cb\u003e19 Mathematics for the cytogenetic technologist 937\u003cbr\u003e\u003c\/b\u003e\u003ci\u003ePatricia K. Dowling\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e19.1 General concepts 937\u003c\/p\u003e \u003cp\u003e19.2 Solutions 942\u003c\/p\u003e \u003cp\u003e19.3 Statistical tools 956\u003c\/p\u003e \u003cp\u003e19.4 Using a hemacytometer 968\u003c\/p\u003e \u003cp\u003e19.5 Quantification and purity determination of DNA using spectroscopy 973\u003c\/p\u003e \u003cp\u003eReference 974\u003c\/p\u003e \u003cp\u003eAdditional readings 974\u003c\/p\u003e \u003cp\u003e\u003cb\u003e20 Selected topics on safety, equipment maintenance, and compliance for the cytogenetics laboratory 975\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eHelen Jenks and Janet Krueger\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e20.1 Introduction 975\u003c\/p\u003e \u003cp\u003e20.2 Biological hazard safety 975\u003c\/p\u003e \u003cp\u003e20.3 Chemical safety 980\u003c\/p\u003e \u003cp\u003e20.4 Fire safety 986\u003c\/p\u003e \u003cp\u003e20.5 Electrical safety 987\u003c\/p\u003e \u003cp\u003e20.6 Disaster plan 988\u003c\/p\u003e \u003cp\u003e20.7 Equipment operation, maintenance, and safety 988\u003c\/p\u003e \u003cp\u003e20.8 Ergonomics 996\u003c\/p\u003e \u003cp\u003e20.9 Regulatory considerations 998\u003c\/p\u003e \u003cp\u003eAcknowledgments 1001\u003c\/p\u003e \u003cp\u003eReferences 1001\u003c\/p\u003e \u003cp\u003eContributed protocols section 1003\u003c\/p\u003e \u003cp\u003eProtocol 20.1 Autoclave sterilization, liquid nitrogen, pro‐par 1003\u003c\/p\u003e \u003cp\u003eProtocol 20.2 Dishwashing procedure 1003\u003c\/p\u003e \u003cp\u003eProtocol 20.3 Eppendorf pipette calibration 1004\u003c\/p\u003e \u003cp\u003eProtocol 20.4 NIST thermometer calibration 1006\u003c\/p\u003e \u003cp\u003eProtocol 20.5 Thermometer calibration 1008\u003c\/p\u003e \u003cp\u003eProtocol 20.6 Timer calibration 1008\u003c\/p\u003e \u003cp\u003e\u003cb\u003e21 A system approach to quality 1011\u003cbr\u003e\u003c\/b\u003e\u003ci\u003ePeggy J. Stupca and Sheryl A. Tran\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e21.1 Quality system 1011\u003c\/p\u003e \u003cp\u003e21.2 Process management 1013\u003c\/p\u003e \u003cp\u003e21.3 Documents and records 1015\u003c\/p\u003e \u003cp\u003e21.4 Assessments 1018\u003c\/p\u003e \u003cp\u003e21.5 Continual improvement 1022\u003c\/p\u003e \u003cp\u003e21.6 Summary 1023\u003c\/p\u003e \u003cp\u003eReferences 1023\u003c\/p\u003e \u003cp\u003eContributed protocols section 1025\u003c\/p\u003e \u003cp\u003eProtocol 21.1 Quality control overview document 1025\u003c\/p\u003e \u003cp\u003eProtocol 21.2 Monitoring specimen quality from off‐hill sites 1030\u003c\/p\u003e \u003cp\u003e\u003cb\u003e22 Laboratory management 1031\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eMervat S. Ayad and Adam Sbeiti\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e22.1 Introduction 1031\u003c\/p\u003e \u003cp\u003e22.2 Management concepts and functions 1032\u003c\/p\u003e \u003cp\u003e22.3 Personnel management 1033\u003c\/p\u003e \u003cp\u003e22.4 Quality management and control 1036\u003c\/p\u003e \u003cp\u003e22.5 Budget development and monitoring 1039\u003c\/p\u003e \u003cp\u003e22.6 Conclusion 1043\u003c\/p\u003e \u003cp\u003eReferences 1043\u003c\/p\u003e \u003cp\u003eSuggested reading 1043\u003c\/p\u003e \u003cp\u003e\u003cb\u003e23 Laboratory information system 1045\u003cbr\u003e\u003c\/b\u003e\u003ci\u003ePeining Li and Richard Van Rheeden\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e23.1 Historical perspective 1045\u003c\/p\u003e \u003cp\u003e23.2 General description of LIS 1045\u003c\/p\u003e \u003cp\u003e23.3 LIS in cytogenetics laboratories 1048\u003c\/p\u003e \u003cp\u003e23.4 Trends for the future LIS 1051\u003c\/p\u003e \u003cp\u003eAcknowledgments 1052\u003c\/p\u003e \u003cp\u003eReferences 1052\u003c\/p\u003e \u003cp\u003e\u003cb\u003e24 Animal cytogenetics 1055\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eMarlys L. Houck, Teri L. Lear and Suellen J. Charter\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e24.1 Introduction 1055\u003c\/p\u003e \u003cp\u003e24.2 Domestic animal fertility 1056\u003c\/p\u003e \u003cp\u003e24.3 Captive management 1057\u003c\/p\u003e \u003cp\u003e24.4 Wildlife conservation 1059\u003c\/p\u003e \u003cp\u003e24.5 General sample collection considerations 1060\u003c\/p\u003e \u003cp\u003e24.6 Fibroblast cell culture 1062\u003c\/p\u003e \u003cp\u003e24.7 Peripheral blood culture 1063\u003c\/p\u003e \u003cp\u003e24.8 Chromosome analysis 1064\u003c\/p\u003e \u003cp\u003e24.9 Molecular and comparative cytogenetics 1070\u003c\/p\u003e \u003cp\u003eAcknowledgments 1071\u003c\/p\u003e \u003cp\u003eGlossary 1072\u003c\/p\u003e \u003cp\u003eReferences 1072\u003c\/p\u003e \u003cp\u003eContributed protocol section 1078\u003c\/p\u003e \u003cp\u003eProtocol 24.1 Blood feather collection 1078\u003c\/p\u003e \u003cp\u003eProtocol 24.2 Avian lymphocyte culture (for large birds) 1078\u003c\/p\u003e \u003cp\u003eProtocol 24.3 Lymphocyte culture using whole blood 1084\u003c\/p\u003e \u003cp\u003eProtocol 24.4 Lymphocyte culture using autologous plasma\/buffy coat (AP\/BC) 1085\u003c\/p\u003e \u003cp\u003eProtocol 24.5 Horse lymphocyte culture method 1087\u003c\/p\u003e \u003cp\u003eProtocol 24.6 Rhino blood culture 1089\u003c\/p\u003e \u003cp\u003eProtocol 24.7 Organ tissue collection protocol from carcass 1090\u003c\/p\u003e \u003cp\u003eProtocol 24.8 Skin biopsy procedure 1090\u003c\/p\u003e \u003cp\u003eProtocol 24.9 Placenta biopsy procedure 1091\u003c\/p\u003e \u003cp\u003eProtocol 24.10 Freezing of fibroblast cell cultures 1092\u003c\/p\u003e \u003cp\u003eProtocol 24.11 Freezing tissue biopsy samples for later initiation of cell culture (tissue piecing) 1094\u003c\/p\u003e \u003cp\u003eProtocol 24.12 Preparation of primary cultures from feather pulp 1095\u003c\/p\u003e \u003cp\u003eProtocol 24.13 Preparation of primary cultures from solid tissue (explants) 1096\u003c\/p\u003e \u003cp\u003eProtocol 24.14 Preparation of primary cultures using enzyme digestion 1097\u003c\/p\u003e \u003cp\u003eProtocol 24.15 Harvesting of fibroblast cell cultures 1098\u003c\/p\u003e \u003cp\u003eProtocol 24.16 Preparation of competitor DNA for FISH hybridization 1099\u003c\/p\u003e \u003cp\u003eProtocol 24.17 In situ hybridization of BAC clones labeled with spectrum fluorochromes:\u003c\/p\u003e \u003cp\u003e probe and slide preparation 1100\u003c\/p\u003e \u003cp\u003eProtocol 24.18 Labeling DNA with spectrum fluorochromes 1102\u003c\/p\u003e \u003cp\u003e\u003cb\u003e25 Online genetic resources and references 1103\u003cbr\u003e\u003c\/b\u003e\u003ci\u003eWahab A. Khan\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e25.1 Introduction 1103\u003c\/p\u003e \u003cp\u003e25.2 Resource information 1103\u003c\/p\u003e \u003cp\u003eIndex 1113\u003c\/p\u003e \u003cp\u003e\u003cb\u003eAbout the Editors\u003c\/b\u003e\u003cbr\u003e\u003cb\u003eMarilyn S. Arsham,\u003c\/b\u003e (retired) Cytogenetic Technologist II, Western Connecticut Health Network, Danbury Hospital campus, Danbury, Connecticut, USA.\u003c\/p\u003e \u003cp\u003e\u003cb\u003eMargaret J. Barch,\u003c\/b\u003e (formerly) Frank F Yen Cytogenetics Laboratory, Weisskopf Child Evaluation Center, University of Louisville, USA.\u003c\/p\u003e \u003cp\u003e\u003cb\u003eHelen J. Lawce,\u003c\/b\u003e Clinical Cytogenetics, Oregon Health \u0026amp; Science University Knight Diagnostics Laboratory, USA.\u003c\/p\u003e \u003cp\u003eCytogenetics is a segment of the evolving cytogenomic branch of genetics that explores the genetic makeup of a cell, in particular chromosomes both in metaphase and in interphase cycles, as well as at the molecular level. It utilizes an assortment of procedures to illuminate both the full complement of chromosomes and a targeted region within a specific chromosome. These investigative tools include routine analysis of G-banded chromosomes, specialized stains that address specific chromosomal structures, and molecular probes, such as fluorescence in situ hybridization and chromosome microarray analysis, which employ a variety of methods to highlight a region as small as a single, specific genetic sequence under investigation.\u003c\/p\u003e  \u003cp\u003eThe AGT Cytogenetics Laboratory Manual, Fourth Edition offers a comprehensive treatment of the diagnostic tests offered by this clinical laboratory environment and explains the scientific theories behind those procedures. One of the most valuable assets is its rich compilation of laboratory-tested protocols currently being used in leading laboratories, along with practical advice for nearly every area of interest to cytogeneticists. In addition to covering essential topics that have been the backbone of cytogenetics for over 60 years, such as the basic components of a cell, use of a microscope, human tissue processing for cytogenetic analysis (prenatal, constitutional, and neoplastic), laboratory safety, and the mechanisms behind chromosome rearrangement and aneuploidy, this edition introduces new and expanded chapters that reflect the intuitive experience of its authors. Some of these new topics include a unique collection of chromosome heteromorphisms; clinical examples of genomic imprinting; an example-driven overview of chromosomal microarray; mathematics specifically geared for the cytogeneticist; usage of ISCN's cytogenetic language; tips for laboratory management; examples of laboratory information systems; a collection of Internet and library resources; and a special chapter on animal chromosomes for the research and zoo cytogeneticist. The range of topics is thus broad yet comprehensive, offering the student with a resource that teaches the procedures performed in the cytogenetics laboratory environment, and the laboratory professional with a peer-reviewed reference that explores the multiple facets behind each of these procedures. Topic coverage is rich in detail, yet presented with the student in mind, making it a useful resource for researchers, clinicians, and laboratory professionals, as well as students in a university or medical school that offers introductory or laboratory-related courses exploring cytogenetics' role within medical genetics.  \u003c\/p\u003e\u003cp\u003eReviews for the third edition:\u003cbr\u003e \"A collection of protocols and explanations for cytogenetic techniques from the common to the peculiar.\"  \u003c\/p\u003e\u003cp\u003e\"An excellent book. Looking forward to an updated version.\"\u003c\/p\u003e","brand":"Wiley-Blackwell","offers":[{"title":"Default Title","offer_id":47990154232037,"sku":"NP9781119061229","price":251.95,"currency_code":"USD","in_stock":false}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1842\/7735\/files\/9781119061229.jpg?v=1761786709","url":"https:\/\/k12savings.com\/products\/the-agt-cytogenetics-laboratory-manual-isbn-9781119061229","provider":"K12savings","version":"1.0","type":"link"}