Techniques in animal cytogenetics:
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English French |
| Veröffentlicht: |
Berlin [u.a.]
Springer [u.a.]
2000
|
| Schriftenreihe: | Principles and practice
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XXVI, 229 S. Ill., graph. Darst. |
| ISBN: | 3540667377 |
Internformat
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| 130 | 0 | |a Techniques de cytogénétique animale | |
| 245 | 1 | 0 | |a Techniques in animal cytogenetics |c Paul Popescu ... (ed.). [Transl. from the French ed. by Ruxandra Popescu] |
| 264 | 1 | |a Berlin [u.a.] |b Springer [u.a.] |c 2000 | |
| 300 | |a XXVI, 229 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 0 | |a Principles and practice | |
| 650 | 7 | |a Cytogénétique - Manuels de laboratoire |2 ram | |
| 650 | 4 | |a Cytogenetic Analysis |v Laboratory Manuals | |
| 650 | 4 | |a Cytogenetics |v Laboratory manuals | |
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Datensatz im Suchindex
| _version_ | 1804127903210799104 |
|---|---|
| adam_text | Titel: Techniques in animal cytogenetics
Autor: Popescu, Paul
Jahr: 2000
Contents
I PREPARATION OF CHROMOSOME SPREADS .... 1
1.1 Cell culture techniques
H. Hayes, B. Dutrillaux ........................... 1
1.1.1 Lymphocyte culture................................. 1
Principle ........................................ 1
Protocol for whole blood .......................... 1
Protocol for lymphocytes isolated by sedimentation .... 2
Protocol for lymphocyte isolation by density
gradient centrifugation .......................... 2
Remarks ......................................... 2
1.1.2 Fibroblast cultures .................................. 3
1.1.2.1 Cultures established from tissue fragments ............. 3
Principle ......................................... 3
Protocol ......................................... 3
Remarks ......................................... 5
1.1.2.2 Initiation of cell cultures from bird embryos ........... 5
Principle ......................................... 5
Protocol ......................................... 5
Remarks ......................................... 5
1.1.2.3 Cell counting ....................................... 5
1.1.2.4 Cell storage......................................... 6
Principle ......................................... 6
Protocol ......................................... 7
1.2 Preparation of chromosomes in prophase
or prometaphase
H. Hayes, B. Dutrillaux ........................... 7
1.2.1 Single or double synchronisation using thymidine ...... 8
Principle ......................................... 8
Protocol for lymphocytes (single synchronisation) .... 8
Protocol for fibroblasts (double synchronisation) ..... 8
Remarks ......................................... 10
1.2.2 Synchronisation by amethopterin ..................... 10
Principle ......................................... 10
Protocol ......................................... 10
Remarks ......................................... 11
1.2.3 Synchronisation by 5-fluorodeoxiuridine
XVIII Contents
P. POPESCU, B. DUTRILLAUX.......................... 11
Principle ......................................... 11
1.2.4 Synchronisation by 5-bromodeoxyuridine or BrdU...... 11
Principle ......................................... 11
1.3 Direct techniques and very short term
development cultures ............................ 12
1.3.1 Study of the bone marrow............................ 12
Protocol ......................................... 12
Remarks ......................................... 12
1.3.2 Study of bird chromosomes from feather pulp.......... 12
Protocol ......................................... 13
Remarks ......................................... 13
1.3.3 Study of fish chromosomes........................... 13
Protocol for culture of blood lymphocytes ........... 14
Protocol for culture of lymphocytes from kidney
or spleen....................................... 14
Protocol for chromosomal banding by BrdU
incorporation in live fish......................... 15
Remarks ......................................... 15
1.3.4 Study of insect chromosomes......................... 16
1.3.4.1 Preparation of metaphase spreads from embryo cells .... 16
Protocol ......................................... 16
1.3.4.2 Preparation of metaphase spreads from gonads ......... 16
Protocol ......................................... 17
1.3.4.3 Chromosome analysis from other tissus ............... 17
1.3.4.4 General comments .................................. 17
1.4 Preparation of chromosome spreads
H. Hayes........................................... 18
Principle ......................................... 18
Protocol ___..................................... 18
Protocol for wet slides ............................. 19
Protocol for dry slides ............................. 19
Remarks ......................................... 19
1.5 Staining techniques of chromosome spreads
H. Hayes, B. Dutrillaux............................ 20
1.5.1 Classical staining using Giemsa....................... 20
Principle ......................................... 20
Protocol ......................................... 20
Remarks ......................................... 20
1.5.2 Orcein staining ..................................... 20
Protocol 1 ........................................ 21
Protocol 2 ........................................ 21
Remarks ......................................... 21
1.5.3 Acridine orange staining............................. 22
Protocol ......................................... 22
Remarks ......................................... 22
1.5.4 Propidium iodide or DAPI staining................... 24
Contents XIX
II CHROMOSOME BANDING TECHNIQUES...... 25
II. 1 Introduction
H. Hayes........................................... 25
11.1.1 Chromosome organisation ........................... 25
11.1.2 Euchromatin ....................................... 27
Distribution of SINE and LINE sequences along
chromosomes .................................. 28
Differential arrangement of AT alignment in Q/G
and R bands.................................... 28
11.1.2.1 Structural bands .................................. 29
Q bands.......................................... 29
G bands.......................................... 30
R bands.......................................... 30
11.1.2.2 Dynamic bands ................................... 31
11.1.3 Code of chromosome banding techniques.............. 33
II.2 Techniques based on DNA structure
B. Dutrillaux, H. Hayes ........................... 33
11.2.1 QFQ bands using quinacrine mustard ................. 34
Principle ......................................... 34
Protocol ......................................... 34
Remarks ......................................... 34
11.2.2 GTG banding by trypsin ............................. 34
Principle ......................................... 34
Protocol ......................................... 34
Remarks ......................................... 34
11.2.3 GAG banding by denaturation ...................... 36
Principle ......................................... 36
Protocol Acid-Saline-Giemsa ....................... 37
Protocol Alkaline-Saline-Giemsa .................... 37
Remarks ......................................... 37
11.2.4 RHG banding by thermal denaturation ................ 37
Principle ......................................... 37
Protocol ......................................... 38
Remarks ......................................... 38
11.2.5 T bands (terminal) .................................. 39
Protocol ......................................... 39
Remarks ......................................... 41
11.2.6 Bands rich in 5-methylcytocine
C. Bourgeois........................................ 41
11.2.6.1 Introduction........................................ 41
11.2.6.2 Immunofluorescent revelation of 5-mC rich bands ...... 43
Principle ......................................... 43
Protocol ......................................... 44
Protocol of denaturation using hydrochloric acid ..... 44
Protocol of denaturation using ultraviolet lamp
radiation....................................... 44
Remarks ......................................... 45
XX Contents
11.3 Banding techniques based on DNA replication
B. Dutillaux, H. Hayes............................. 46
11.3.1 R or G bands by incorporation of BrdU................ 46
Principle ......................................... 46
11.3.1.1 Immunoflorescent detection of BrdU incorporation
in chromosomes .................................... 48
Principle ......................................... 48
Protocol ......................................... 48
Remarks ......................................... 49
11.3.1.2 FPG staining technique (fluorochrome-photolysis-
Giemsa)............................................ 49
Principle ......................................... 49
Protocol ......................................... 51
Remarks ......................................... 51
11.3.1.3 Propidium iodide staining technique .................. 51
Principle ......................................... 51
Protocol ......................................... 51
Remarks ......................................... 52
11.3.1.4 DAPI staining technique ............................. 53
Principle ......................................... 53
Protocol ......................................... 53
11.3.1.5 Preparation of chromosomes labeled with BrdU during
the second half of the S phase to produce R bands ...... 54
Protocol ......................................... 54
Remarks ......................................... 54
11.3.1.6 Preparation of chromosomes labeled with BrdU during
the first half of the S phase to produce G bands ......... 55
Protocol ......................................... 55
Remarks ......................................... 55
11.3.2 Sister chromatid exchanges (SCE) ..................... 55
Principle ......................................... 55
Protocol ......................................... 55
Remarks ......................................... 56
11.3.3 Asymmetrical incorporation of BrdU.................. 56
11.4 Techniques of chromosome differentiation
based on DNA base composition
B. Dutrillaux..................................... 57
II.4.1 Treatment by 5-azacytidine or 5-azadeoxycytidine....... 57
Principle ......................................... 57
Protocol ......................................... 57
Remarks ......................................... 57
11.5 Heterochromatin staining ....................... 58
H.5.1 CBG bands ......................................... 58
Principle ......................................... 58
Protocol ......................................... 58
Remarks ......................................... 58
Contents XXI
11.5.2 CT bands........................................... 61
Principle ......................................... 61
Protocol ......................................... 61
11.5.3 Gil bands.......................................... 61
Principle ......................................... 61
Protocol ......................................... 61
Remarks ......................................... 62
11.5.4 DA-DAPI staining ................................... 62
Principle ......................................... 62
Protocol ......................................... 64
Remarks ......................................... 64
11.6 Staining of nucleolar organiser regions NOR
H. Hayes, B. Dutrillaux............................ 65
Protocol ......................................... 65
Remarks ......................................... 65
11.7 Techniques of sequential banding
B. Dutrillaux..................................... 65
Principle ......................................... 65
Protocol of sequential Q, R, and C banding .......... 68
Protocol of sequential Q and NOR banding, or R
and NOR banding .............................. 68
III IN SITU HYBRIDISATION TECHNIQUES
H. Hayes, B. Dutrillaux............................ 69
111.1 Introduction ..................................... 69
111.1.1 In situ hybridisation using radioactive probes .......... 69
III. 1.2 In situ hybridisation using non radioactive probes ...... 70
HI. 1.3 Identification of hybridised chromosomes ............. 71
111.2 Methods.......................................... 72
IH.2.1 Preparation of chromosome spreads................... 72
111.2.2 DNA probe labelling ................................ 73
IH.2.2.1 Non radioactive labelling of long DNA probes ( lkb) ___ 73
Principle ......................................... 73
Protocol for nick translation labelling ............... 73
HI.2.2.2 Non radioactive labelling of short DNA probes
(00.25-1.5kb) ....................................... 74
Principle ......................................... 74
Protocol ......................................... 74
Remarks ......................................... 74
III.2.2.3 Radioactive labelling ................................ 75
Protocol for the use of the nick translation method ___ 75
IH.2.3 Pretreatment of the chromosome preparations using
ribonuclease A...................................... 75
Principle ......................................... 75
Protocol ......................................... 75
XXII Contents
111.2.4 Chromosomal DNA denaturation ..................... 75
Principle ......................................... 75
Protocol ......................................... 76
111.2.5 Probe preparation, labelling and denaturation.......... 76
111.2.6 In situ hybridisation................................. 76
111.2.7 Posthybridisation washes ............................ 76
111.2.7.1 Radioactive probes .................................. 76
111.2.7.2 Non radioactive probes .............................. 76
111.2.8 Hybridisation signal detection and banding............ 77
111.2.8.1 Radioactive probes: autoradiographic detection ......... 77
Principle ......................................... 77
Protocol ......................................... 78
111.2.8.2 Non radioactives probes: immunoreaction detection..... 79
Protocol ......................................... 79
IH.2.9 Microscopy and photography......................... 79
III.2.9.1 Radioactive probes .................................. 79
HI.2.9.2 Non radioactives probes.............................. 79
III.3 Remarks on other applications of in situ
hybridisation...................................... 80
IV METHODS OF GERM CELLS STUDY ............ 85
IV. 1 Meiosis in male
P. Popescu.......................................... 85
IV.1.1 Classical method.................................... 85
Protocol ......................................... 85
Remarks ......................................... 86
I V.I.2 Synaptonemal complex method
Y. Rumpler, O. Gabriel-Robez...................... 86
Principle ......................................... 88
Protocol ......................................... 89
Remarks ......................................... 92
IV.2 Meiosis in the mammalian female
J.F. Ectors, L. Koulisher............................ 94
Principle ......................................... 94
Protocol ......................................... 95
IV.3 Study of the spermatozoa by interspecific in
vitro fertilisation (insemination)
P. Popescu.......................................... 97
Principle ......................................... 97
Protocol ......................................... 97
Protocol of capacitation in the cold state ............. 99
Protocol of capacitation in the warm state ........... 99
Remarks ......................................... 102
Contents XXIII
V THE LAMPBRUSH CHROMOSOMES OF
AMPHIBIANS
F. Simon, N. Vichniakova, C. payne, J.C. Lacroix...... 103
V.I Introduction: The basis of lampbrush
chromosomes mapping .......................... 103
V.I.I Organisation of the lampbrush chromosomes........... 103
V. 1.2 Morphologic mapping of the lampbrush
chromosomes....................................... 107
V.I.2.1 Morphological variations of genetic origin ............. 107
V. 1.2.2 Morphological variations of physiological origin ........ 109
V.1.3 Immunomorphological mapping...................... 109
V.1.4 Maps of the lampbrush chromosomes
of Pleurodeles ...................................... 112
V.l.4.1 The conditions of map development................... 112
V.l.4.2 The intraspecific maps ............................... 112
V.2 Technique for the preparation of lampbrush
chromosomes for light microscopy.............. 114
V.2.1 Preparation of oocytes............................... 114
Protocol ......................................... 114
V.2.2 Chromosome preparation for morphological
studies............................................. 115
Protocol ......................................... 115
V.2.3 Chromosome immunolabelling ....................... 118
Protocol ......................................... 118
V.3 Preparation of lampbrush chromosomes
for electron microscopy.......................... 120
V.3.1 Ultrastructural studies............................... 120
Protocol ......................................... 121
V.3.2 High resolution immunolabelling...................... 121
Principle ......................................... 121
V.3.2.1 Pre-embedding immunolabelling...................... 123
Protocol ......................................... 123
V.3.2.2 Post-embedding immunolabelling..................... 124
Protocol ......................................... 124
V.4 Preparation of mitotic chromosomes............ 124
Protocol ......................................... 125
V.5 Analysis of lampbrush chromosomes ........... 125
V.5.1 Morphological markers .............................. 128
V.5.1.1 Axial structures ..................................... 128
V.5.1.2 Loops.............................................. 129
V.5.2 Immunolabelling.................................... 130
V.5.3 Mapping parameters................................. 131
V.5.3.1 Chromosome classification .......................... 131
V.5.2.2 Chromosome orientation ............................ 131
V.5.3.3 Marker localisation ................................. 131
XXIV Contents
V.5.3.4 Correspondences between the lampbrush chromosome
maps............................................... 131
V.6 The importance of the lampbrush
chromosomes ................................... 133
V.6.1 Detection and analysis of chromosomal
rearrangements..................................... 133
V.6.2 Study of populations and evolution of chromosomes .... 133
V.6.3 Study of the transcriptional physiology in situ.......... 136
VI TECHNIQUES FOR THE SIUDY OF
DROSOPHILA CHROMOSOMES
F. Lemeunier, S. Aulard............................ 137
VI. 1 Mitotic chromosomes............................ 137
VI.l.l Introduction........................................ 137
VI.1.2 Chromosome spreads preparation..................... 138
Principle ......................................... 138
Protocol ......................................... 138
VI. 1.3 Staining and banding of mitotic chromosomes:
remarks............................................ 139
VI. 1.4 In situ hybridisation................................. 140
VI.2 Polythene chromosomes ......................... 140
VI.2.1 Introduction........................................ 140
VI.2.2 Structure........................................... 141
VI.2.3 Reference maps ..................................... 142
VI.2.4 Chromosome preparation: Classical and molecular
cytogenetic technique ............................... 145
VI.2.4.1 Breeding conditions of Drosophila .................... 145
VI.2.4.2 Chromosome preparation: classical analysis ............ 145
Protocol ........................................... 145
VI.2.4.3 Chromosome preparation: in situ hybridisation ........ 146
Principle .......................................... 146
Protocol .......................................... 146
VI.2.5 In situ hybridisation.................................. 147
Protocol ........................................... 147
VI.2.6 Chromosome observation ............................ 149
VII TECHNIQUES FOR THE STUEY OF
INTERPHASE NUCLEUS ....................... 151
VII. 1 Sex chromatin examination
P. Popescu .......................................... 151
Principle .......................................... 151
Protocol .......................................... 151
Remarks .......................................... 151
VII.2 Released chromatin (Chromatin halo) ........... 152
Protocol ........................................... 152
Contents XXV
VII.3 In situ hybridisation of interphasic nuclei
C. Bourgeois........................................ 153
Principle .......................................... 153
Protocol .......................................... 153
Protocol for paraffin sections........................ 153
Protocol for nucleus and spread cells or slides
with cell layers................................... 154
Remarks .......................................... 155
VIII APPLICATION OF FLOW CYTOMETRY
AND SLIT-SCAN FLUOROMETRY IN
ANALYSIS AND SORTING OF MAMMALIAN
CHROMOSOMES
M. Haussmann, C. Cremer........................... 157
VIII.1 Introduction ..................................... 157
VIII.2 Principles of the flow cytometry ................. 158
VIII.2.1 Standard flow cytometry.............................. 158
VIII.2.2 Slit scan fluorometry.................................. 159
VIII.2.3 Flow sorting ......................................... 161
VIII.2.4 Computing........................................... 161
VIII.3 Methods of chromosome preparation and
staining......................................... 161
VIII.3.1 Modified hexanediole method ......................... 162
References ......................................... 162
Protocol ........................................... 162
Remarks........................................... 163
VIII.3.2 TAcCaM - method.................................... 163
References ......................................... 163
Protocol ........................................... 163
Remarks .......................................... 163
VIII.3.3 Methanol - acetic acid method......................... 164
References ......................................... 164
Protocol ........................................... 164
Remarks........................................... 164
VIII.3.4 Tris/MgCl2/Triton X10000 method ..................... 164
References ......................................... 164
Protocol ........................................... 164
VIII.3.5 Polyamine method.................................... 164
References ......................................... 164
Protocol ........................................... 164
VIII.3.6 Modified polyamine method .......................... 165
References ......................................... 165
Protocol ........................................... 165
VIII.3.7 HEPES/MgSO4 method................................ 165
References ......................................... 165
Protocol ........................................... 165
XXVI Contents
VHI.3.8 Modified HEPES method.............................. 166
References......................................... 166
Protocol ........................................... 166
Remarks........................................... 166
VIII.3.9 Fluorescein labelling (FITC) by in situ hybridisation
suspension........................................... 166
References ......................................... 166
Protocol ........................................... 166
Remarks........................................... 167
VIII.3.10 Dyes and lasers...................................... 167
VIII.4 Measurements and evaluation in flow
cytometry....................................... 167
VIII.4.1 Flow karyotypes ..................................... 167
VIII.4.2 Data evaluation of flow karyotypes..................... 171
VIII.4.3 Slit scan measurements ............................... 172
VIII.4.4 Perspectives.......................................... 175
VIII.4.5 Identification of the sorted chromosomes by GTG
banding
P. Popescu .......................................... 176
Principle .......................................... 176
Protocol ........................................... 177
Remarks .......................................... 177
VIII.5 In situ hybridisation to chromosomes
in suspension
D. Celeda ........................................... 178
Principle .......................................... 178
Protocol .......................................... 180
Remarks .......................................... 186
Appendix...................................................... 187
S Solutions for chromosome staining and banding
techniques ............................................... 187
M Miscellaneous ............................................ 189
L Solutions for lampbrush chromosomes ..................... 192
F Solutions for flow and slit scan cytometry................... 193
H Solutions for in situ hybridisation.......................... 196
CM Culture media............................................ 197
SS Stock solutions ........................................... 199
B Buffer solutions........................................... 201
V Solutions for in vivo treatments............................ 203
Glossary ...................................................... 205
References..................................................... 209
Index ......................................................... 225
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| id | DE-604.BV013202437 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T18:40:44Z |
| institution | BVB |
| institution_GND | (DE-588)35926-9 |
| isbn | 3540667377 |
| language | English French |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-008995331 |
| oclc_num | 43684521 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-M49 DE-BY-TUM DE-20 |
| owner_facet | DE-355 DE-BY-UBR DE-M49 DE-BY-TUM DE-20 |
| physical | XXVI, 229 S. Ill., graph. Darst. |
| publishDate | 2000 |
| publishDateSearch | 2000 |
| publishDateSort | 2000 |
| publisher | Springer [u.a.] |
| record_format | marc |
| series2 | Principles and practice |
| spelling | Techniques de cytogénétique animale Techniques in animal cytogenetics Paul Popescu ... (ed.). [Transl. from the French ed. by Ruxandra Popescu] Berlin [u.a.] Springer [u.a.] 2000 XXVI, 229 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Principles and practice Cytogénétique - Manuels de laboratoire ram Cytogenetic Analysis Laboratory Manuals Cytogenetics Laboratory manuals Tierzelle (DE-588)4185509-7 gnd rswk-swf Cytogenetik (DE-588)4070176-1 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Tierzelle (DE-588)4185509-7 s Cytogenetik (DE-588)4070176-1 s Methode (DE-588)4038971-6 s DE-604 Popescu, Paul 1936- Sonstige (DE-588)121763005 oth Institut national de la recherche agronomique (Paris) Sonstige (DE-588)35926-9 oth HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008995331&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Techniques in animal cytogenetics Cytogénétique - Manuels de laboratoire ram Cytogenetic Analysis Laboratory Manuals Cytogenetics Laboratory manuals Tierzelle (DE-588)4185509-7 gnd Cytogenetik (DE-588)4070176-1 gnd Methode (DE-588)4038971-6 gnd |
| subject_GND | (DE-588)4185509-7 (DE-588)4070176-1 (DE-588)4038971-6 |
| title | Techniques in animal cytogenetics |
| title_alt | Techniques de cytogénétique animale |
| title_auth | Techniques in animal cytogenetics |
| title_exact_search | Techniques in animal cytogenetics |
| title_full | Techniques in animal cytogenetics Paul Popescu ... (ed.). [Transl. from the French ed. by Ruxandra Popescu] |
| title_fullStr | Techniques in animal cytogenetics Paul Popescu ... (ed.). [Transl. from the French ed. by Ruxandra Popescu] |
| title_full_unstemmed | Techniques in animal cytogenetics Paul Popescu ... (ed.). [Transl. from the French ed. by Ruxandra Popescu] |
| title_short | Techniques in animal cytogenetics |
| title_sort | techniques in animal cytogenetics |
| topic | Cytogénétique - Manuels de laboratoire ram Cytogenetic Analysis Laboratory Manuals Cytogenetics Laboratory manuals Tierzelle (DE-588)4185509-7 gnd Cytogenetik (DE-588)4070176-1 gnd Methode (DE-588)4038971-6 gnd |
| topic_facet | Cytogénétique - Manuels de laboratoire Cytogenetic Analysis Laboratory Manuals Cytogenetics Laboratory manuals Tierzelle Cytogenetik Methode |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008995331&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | UT techniquesdecytogenetiqueanimale AT popescupaul techniquesinanimalcytogenetics AT institutnationaldelarechercheagronomiqueparis techniquesinanimalcytogenetics |