Analyzing Chromosomes

Header......Page 1
Cover......Page 2
Contents......Page 7
Abbreviations......Page 11
Contributors......Page 13
Introduction......Page 15
2. The cytoplasm......Page 17
3. The nucleus......Page 19
4.1 RNA transcription......Page 21
5. Mutation and DNA repair......Page 23
6. Chromosome organization and structure......Page 24
6.1 Mitosis......Page 27
6.2 Meiosis......Page 29
7.1 Chromosome architecture and transcriptional activity......Page 32
7.2 The difference between the X and Y chromosomes of mammals......Page 33
Further reading......Page 34
2.2 Blood cell culture introduction......Page 35
3.1 Receipt of the sample......Page 36
3.4 Alternative harvest techniques......Page 37
5.3 Setting up of cultures......Page 38
6.2 Introduction to bone marrow, leukemic blood, and lymph node cell culture......Page 39
6.3 Setting up tissues from malignant conditions......Page 40
7.2 Culture of tissues from chornic lymphoid leukemias and lymphomas -- introduction......Page 41
Further Reading......Page 42
Protocol 2.1 - Setting up cell cultures and harvesting......Page 43
Protocol 2.2 - Amniotic fluid culture......Page 45
Protocol 2.3 - Harvesting amniocytes......Page 46
Protocol 2.4 - Thymidine blocking......Page 48
Protocol 2.5 - Addition of BrdU with colcemid......Page 49
Protocol 2.6 - Sorting of the chorionic villus sample......Page 50
Protocol 2.7 - Direct culture of chorionic villus samples......Page 51
Protocol 2.8 - Long-term culture of chorionic villus samples......Page 53
Protocol 2.9 - Setting up bone marrow and leukemic blood......Page 56
Protocol 2.10 - Synchronization of bone marrow cultures using a BrdU/FdU cocktail mixture......Page 58
Protocol 2.11 - Harvest of bone marrow and blood cultures......Page 59
Protocol 2.12 - Culture of samples from CLL blood and marrow......Page 61
Protocol 2.13 - Preparation and culture of samples from lymphomas......Page 62
Protocol 2.14 - Slide-making......Page 63
1. Introduction......Page 65
2. Banding techniques and their applications......Page 66
Acknowledgments......Page 67
References......Page 68
Protocol 3.1 - Solid staining......Page 69
Protocol 3.2 - Preparation of Leishman's stain for G-banding......Page 71
Protocol 3.3 - G-banding......Page 72
Protocol 3.4 - Q-banding......Page 75
Protocol 3.5 - C-banding......Page 77
Protocol 3.6 - R-banding......Page 79
Protocol 3.7 - Nucleolar organizer region (NOR) staining......Page 81
Protocol 3.8 - DAPI/distamycin A staining......Page 83
2. Types of chromosomal changes......Page 85
2.1 Numerical chromosomal changes......Page 87
2.2 Structural chromosomal changes......Page 88
2.3 Uniparental diploidy, disomy and genomic imprinting......Page 89
3. Nomenclature for human chromosomes......Page 94
3.1 Nomenclature symbols......Page 96
3.2 Nomenclature symbols for acquired abnormalities and their respective clones......Page 101
References......Page 105
1. Introduction......Page 107
1.1 Methods of analysis......Page 108
2.1 Chromosome changes occuring from conception to term......Page 112
2.2 Chromosome changes occuring in neonates and young children with congenital malformations and/or developmental delay......Page 113
2.4 Other Chromosomal observations......Page 114
2.6 Problems of interpretation arising in cell culture......Page 116
3.1 Prompt arrival of the sample......Page 119
3.3 The number of metaphase spreads to be analyzed......Page 120
3.4 Cytogenetic analysis experience......Page 121
4. Clonal abnormality......Page 122
References......Page 123
1. Introduction......Page 125
2. Trisomy 21 - Down syndrome......Page 127
3. Trisomy 18 - Edward syndrome......Page 128
5. 45,X - Turner syndrome......Page 129
6.1 Clinical findings......Page 130
8. 47, XYY......Page 131
11. Microdeletion syndromes and chromosomal duplications......Page 132
12. Marker chromosomes......Page 133
13. Ring chromosomes......Page 136
14. Fragile X syndrome......Page 137
References......Page 138
Further Reading......Page 139
1. Introduction......Page 141
2.1 Acute myeloid leukemia......Page 142
2.2 Myelodysplastic syndromes (MDS) and myeloproliferative disorders (MPD)......Page 144
3.2 Acute myeloid leukemia - FAB category M1......Page 146
3.3 Acute myeloid leukemia - FAB category M2......Page 147
3.5 Acute promyelocytic leukemia - FAB category M3 and M3V......Page 148
3.7 Acute myeloid leukemia - FAB categories M4 and M5......Page 149
3.8 Acute myeloid leukemia - FAB category M5a......Page 150
3.9 Acute myeloid leukemia - FAB category M6......Page 151
4. Chromosome changes in myelodysplastic syndromes......Page 152
5.1 Chronic myeloid leukemia......Page 154
5.2 Polycythemia rubra vera......Page 156
6.3 Immunophenotyping......Page 157
6.4 Hematological findings and symptoms of disease......Page 158
6.5 Chromosome abnormalities observed in ALL......Page 159
6.6 Other abnormalities in ALL......Page 164
7.2 Established chromosome abnormalities in chronic lymphoproliferative disorders......Page 165
7.3 The lymphomas......Page 167
7.4 Chromosome abnormalities observed in non-Hodgkin's lymphomas......Page 168
8.1 Chromosome changes in solid tumors......Page 171
9. Summary......Page 172
Further Reading......Page 173
2. 1 Introduction......Page 175
2.3 Important steps in FISH......Page 176
2.4 Reporting of FISH results......Page 181
3. Twenty-four color painting......Page 183
4. Color banding......Page 184
7. Comparative genomic hybridization......Page 185
7.1 Overview of the techniques used in CGH......Page 186
8. The present and future of cytogenetics......Page 187
References......Page 188
Protocol 8.1 - Processing of uncultured blood and bone marrow preparations for interphase FISH......Page 189
Protocol 8.2 - Processing of uncultured amniocytes for interphase FISH......Page 190
Protocol 8.3 - Pre-treatment of slides for uncultured samples (or very cytoplasmic samples)......Page 192
Protocol 8.4 - Standard protocol for in situ hybridization......Page 194
Appendix 1 - Glossary of terms......Page 199
Appendix 2......Page 207
Index......Page 215