ISBN: 3-540-64602-7
TITLE: Diagnostic Cytogenetics
AUTHOR: Wegner, Rolf-Dieter (Ed.)
TOC:

Part I Classical Cytogenetics 
Chapter 1 
Tissue Culture 
FRIEDEL WENZEL 3 
Subprotocol 1: Sampling, Transport, Storage 20 
Subprotocol 2: Cryopreservation 26 
Subprotocol 3: Bacterial and/or fungal contamination 33 
Subprotocol 4: Mycoplasma Contamination 37 
Subprotocol 5: Elimination of Mycoplasmas from Cell Cultures 45 
Chapter 2 
Chromosome Staining 
ANGELIKA KHLER 52 
Subprotocol 1: Giemsa Staining 55 
Subprotocol 2: GTG-Banding 56 
Subprotocol 3: QFQ-Banding 59 
Subprotocol 4: C-Banding 60 
Subprotocol 5: NOR-Staining 62 
Subprotocol 6: DA-DAPI Staining 64 
Subprotocol 7: Replication Pattern (by BrdU-Incorporation) 65 
Subprotocol 8: Sister Chromatid Differentiation 
(by BrdU Incorporation) 68 
Subprotocol 9: Sex Chromatin Staining - X Chromatin (Barr Body) 70 
Subprotocol 10: Sex Chromatin Staining - Y-Chromatin (Y Body) 72 
Chapter 3 
Karyotyping and Data Interpretation 
KARSTEN HELD 75 
Chapter 4 
Documentation 
PETER MINY AND ROLF-DIETER WEGNER 96 
Part II Postnatal Diagnosis 
Chapter 5 
Peripheral Blood 
IRIS BARTELS 115 
Chapter 6 
Establishment of Permanent Growing Lymphoblastoid Cell Lines 
HEIDEMARIE NEITZEL 121 
Subprotocol 1: Transformation of B Lymphocytes 
with B95-8 EB Virus 123 
Subprotocol 2: Ficoll Separation of Unfractionated Mononuclear 
Leukocytes Obtained from Whole Blood 125 
Subprotocol 3: Establishment of Cultures 126 
Subprotocol 4: Freezing of Lymphoblastoid Cells 127 
Subprotocol 5: Thawing of Lymphoblastoid Cells 128 
Subprotocol 6: Chromosome Preparations 
from Lymphoblastoid Cells 129 
Chapter 7 
Solid Tissues 
REGINE SCHUBERT AND GESA SCHWANITZ 132 
Chapter 8 
Cells from Urine Sample 
HANNELORE KRNER, HENRIKE DIA AND CHRISTIANE BOMMER 142 
Chapter 9 
Classical and Molecular Cytogenetics of Tumor Cells 
BRIGITTE SCHLEGELBERGER, SIMONE METZKE, SVETLANA HARDER, 
REINA ZHLKE-JENISCH,YANMING ZHANG, REINER SIEBERT 151 
Subprotocol 1: Chromosome Analysis 152 
Subprotocol 2: Fluorescence in Situ Hybridization 168 
Chapter 10 
Cytogenetics of Meiotic Cells 
R. JOHANNISSON 186 
Subprotocol 1: Air-Drying Method 188 
Subprotocol 2: Surface Spreading Method Using Light Microscopy 190 
Subprotocol 3: Surface Spreading Using Electron Microscopy 199 
Subprotocol 4: Ejaculate 207 
Part III Prenatal Diagnosis 
Chapter 11 
Prenatal Diagnosis - An Introduction 
ROLF-DIETER WEGNER 213 
Chapter 12 
Amniotic Fluid Cell Analysis 
INGO KENNERKNECHT, MAHMOUD DJALALI, GOTTHOLD BARBI, 
WALTER JUST AND WALTHER VOGEL 217 
Chapter 13 
Chorionic Villi Sampling 
ROLF-DIETER WEGNER AND HOLGER TOENNIES 231 
Subprotocol 1: Setting-up a Short Term Culture 235 
Subprotocol 2: Chromosome Preparation from Short 
Term Culture 238 
Subprotocol 3: Setting-up a Long Term Culture 
by Physical Maceration 239 
Subprotocol 4: Setting-up a Long Term Culture 
by Enzymatic Dissociation 241 
Subprotocol 5: Cell Harvest and Chromosome Preparation 242 
Part IV Special Applications 
Chapter 14 
Diagnosis of Chromosomal Instability Syndromes 
ROLF-DIETER WEGNER AND MARKUS STUMM 251 
Chapter 15 
Flow Cytometric Testing for Syndromes with Chromosomal 
Instability, Aplastic Anemia and Related Hematological Disorders 
DETLEV SCHINDLER AND HOLGER HOEHN 269 
Chapter 16 
Cell Fusion 
MARKUS STUMM AND ROLF-DIETER WEGNER 282 
Chapter 17 
Origin of Trisomies 
GESA SCHWANITZ AND THOMAS EGGERMANN 291 
Part V Molecular Cytogenetics 
Chapter 18 
Fluorescence in Situ Hybridization 
GESA SCHWANITZ AND REGINE SCHUBERT 305 
Subprotocol 1: Preparation of Slides 310 
Subprotocol 2: Labelling of DNA Probes 313 
Subprotocol 3: Denaturation and Hybridization 317 
Subprotocol 4: Detection of Biotin Labelled Probes 319 
Subprotocol 5: Amplification 322 
Subprotocol 6: Simultaneous Two Colour Detection 323 
Subprotocol 7: In Situ Hybridization with Unique Sequence Probes 324 
Subprotocol 8: Preparation of Interphase Nuclei 326 
Chapter 19 
Fluorescence in Situ Hybridization (FISH) 
Analysis in Human Sperm Cells 
EVELYN KO AND RENE MARTIN 335 
Chapter 20 
Microdissection and Reverse Chromosome Painting 
GABRIELE SENGER, JRG WEIMER, UWE CLAUSSEN, 
AND ILSE CHUDOBA 356 
Chapter 21 
Comparative genomic hybridisation (CGH) 
TRAUDL HENN AND OSKAR A. HAAS 376 
Part VI Techniques in Development 
Chapter 22 
Fetal Cells in Maternal Blood 
DOROTHEE GNSHIRT, HENK S.P. GARRITSEN, 
WOLFGANG HOLZGREVE 401 
Chapter 23 
Spectral Karyotyping in Clinical and Tumor Cytogenetics 
EVELIN SCHRCK, YUVAL GARINI, MICHAEL KHLER 
AND THOMAS RIED 416 
Chapter 24 
Chromosome Analysis by Multiplex-FISH (M-FISH) 
MICHAEL R. SPEICHER 439 
Subject Index 457 
END
