Radioisotopes in biology: a practical approach
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford [u.a.]
Oxford Univ. Press
2002
|
| Ausgabe: | 2. ed. |
| Schriftenreihe: | The practical approach series
252 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XX, 307 S. Ill., graph. Darst. |
| ISBN: | 0199638276 0199638268 |
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Datensatz im Suchindex
| _version_ | 1804128670515724288 |
|---|---|
| adam_text | Contents
Preface page v
Protocol list xv
Abbreviations xix
1 Introduction 1
Robert]. Slater
1 The decision to use a radioisotope ]
2 Laboratory facilities 1
3 The choice of a labelled compound 1
The choice of a radionuclide 2
Labelling position 2
Specific activity 3
The isotope effect 4
4 Storage and purity 4
5 Experimental design 5
2 Radioisotope use 7
David Prime and Barry Frith
1 Introduction 7
2 Radioactivity 7
Radioactive decay 7
Units of radioactivity 10
Properties of radiations 10
Radiation dose and equivalent dose 11
External radiation doses from beta emitters 11
External radiation doses from X rays and gamma rays 12
Internal radiation doses from radionuclides 13
3 Biological effects of radiation and the basis of legislation 14
Effective dose 14
Tissue level effects 14
Carcinogenic effects 15
Genetic effects 15
4 Biological basis of dose limits 16
Risk rates for biological effects 16
5 Working with unsealed sources 18
Getting started 38
Designation of working areas 19
6 Designation of classified workers 20
7 Controlling the risks to radiation workers 20
The non classified worker 20
The classified worker 21
8 The tracer study 21
Experimental design 21
The purchase of isotopes and labelled compounds 23
Receiving the isotope 23
Isotope manipulations 23
Design of isotope laboratory 24
9 Personal protection in the radioisotope laboratory 24
10 Routine monitoring of area 25
External dose measurement 25
Measurement of air contamination 25
Measurement of surface contamination 26
Estimation of individual dose 26
11 Transport of radioactive material 26
Requirements and controls for transport of excepted packages 27
12 Disposal of radioactive waste 28
Solid waste 29
Liquid low level waste (aqueous and approved liquid scintillation cocktails) 29
References 30
3 Risk assessments for the use of radloisotopes in biology 31
Peter E.BaMrtce
1 Introduction 31
2 The Health and Safety at Work etc. Act 1974 32
3 The Management of Health and Safety at Work Regulations 1999 33
4 The risk assessment process 35
The content of the risk assessment 35
Responsibility for making risk assessments 36
5 Suggested methods for making risk assessments 37
General laboratory hazards 37
Use of generic risk assessments in biological research and teaching 38
6 Hazards and risks to health from the use of radioactive substances and
ionizing radiation 46
7 Special hazard and risk assessment for 125I 48
The hazard 48
Risks to health and radiation protection procedures to be followed 48
Elimination of the hazard 50
8 Special hazard and risk assessment for DNA labelling/sequencing work
with 32P 50
Minimizing uptakes 50
Assessment of risks and control of external radiation from 32P 51
9 Risk assessments for gamma emitting radionuclides 55
10 Risk assessments for environmental protection 57
11 Risks to the health of biological research workers in relation to
other risks 58
References 60
4 Radioisotope detection using X ray film 63
RonALaskey
1 Introduction 63
2 Choosing a class of detection method: autoradiography versus
fluorography or intensifying screens 64
Direct autoradiography gives high resolution but limited sensitivity 64
Fluorography increases sensitivity for weak (J emitters 64
Intensifying screens increase sensitivity for high energy p particles or 7 rays 65
3 Direct autoradiography 66
Procedure for direct autoradiography 66
4 Use of X ray intensifying screens to increase sensitivity for 32P and 7 ray
emitters 67
5 Fluorography of weak p emitters 70
General technical procedures for fluorography of acrylamide gels by
all methods 73
Procedures for fluorography of acrylamide gels 72
Fluorography of other samples excluding acrylamide gels 76
6 General technical considerations for radioisotope detection by X ray film 79
Choice of film and screens 79
Conditions for exposure and processing of film 80
Pre exposure of film to hypersensitize it for use with intensifying screens or
fluorography 80
Procedures for drying acrylamide or agarose gels 82
7. The underlying principles of radioisotope detection by X ray film 82
Acknowledgements 83
References 83
5 The scintillation counter 85
Charles LDodson
1 Introduction 85
2 Counting efficiency and counting error 85
3 Counting error 86
4 Origin and nature of nuclear emissions 87
5 Interaction of decay species with matter 89
6 The liquid scintillation counter 91
Introduction 91
The scintillation cocktail 92
Beta particle spectrum 94
The liquid scintillation counter 95
7 Advanced topics 107
Multi label counting 107
Colour quench detection and correction 109
D.p.m. correction without quench curves 130
Sample monitoring 111
8 Special counting situations 111
Cerenkov counting 111
Low level counting 113
9 Solid scintillators and programmable coincidence gates 113
10 Sample preparation 114
Cocktail properties 115
Sample types 116
Vials 120
11 Gamma counters 120
Gamma counter operational parameters 121
12 Flow counters 124
13 Multi detector liquid scintillation and gamma counters 125
14 Storage phosphor technology 125
Phosphor screens 126
Scanning unit 126
Phosphor screen erasure 127
Quantitative analysis 128
References 128
Further reading 129
6 In vitro labelling of nucleic acids and proteins i3i
Martin W. Cunningham, A Patel, A C. Simmonds and D. Williams
1 Introduction 131
Choice of radiolabel 131
2 Applications of nucleic acid labelling 133
3 Labelling methods for hybridization probes 134
Random prime labelling 136
Nick translation 138
Transcription labelling 141
End labelling of oligonucleotides 145
Choosing a radioactive labelling system for nucleic acid probes 151
4 Cycle sequencing with 33P labelled terminators 154
5 Applications of labelled proteins 156
6 Labelling of proteins with 125I 156
General considerations 156
Direct labelling 156
Indirect methods 161
Purification of radio iodinated proteins 162
Determination of specific activity 163
7 Labelling of proteins with 3H and 14C 164
Tritium labelling 164
Carbon 14 labelling 165
8 Protein kinase assays 165
9 In vitro translation with radiolabelled amino acids 168
Acknowledgements 169
References 170
7 Subcellular localization of biological molecules m
Richard Cumming, Ian Durrant and Rachel FaUm
1 Introduction 173
Compartmentalization within the cell 371
2 Subcellular localization of proteins 172
Introduction 172
Western blotting 173
Immunoprecipitation 176
Histological techniques 178
3 Subcellular localization of nucleic acids 181
Identification of nucleic acid molecules isolated from subcellular fractions 181
Localization of nucleic acids using ISH 182
Safety considerations when using ISH (32) 185
Preparation of target cells or tissues 186
Preparation ofradiolabelled probe 190
Hybridization 192
Autoradiography 195
Analysis of data 199
Controls required for ISH 199
Simultaneous localization of proteins and nucleic acids 205
4 Conclusion 205
Acknowledgements 206
References 206
8 Radioisotopes and immunoassay 209
Adrian F. Bristow, Andrew]. H. Gearing and Robin Thorpe
1 Introduction 209
2 Immunoassays: theoretical considerations 210
3 Radioimmunoassays 213
Reagents 213
Assay conditions 219
Separation techniques 219
Specific protocols 221
Interpretation of results 223
4 Immunoradiometric assays 226
Choice of solid support 226
Antibodies for IRMAs 226
Radiolabelling antibodies 227
Assay design 227
5 Non radioisotopic immunoassays 229
Enzyme immunoassays 229
Radioisotopes versus non isotopic methods in immunoassay: future trends 230
References 232
9 Pharmacological techniques 233
Isabel J. M. Beresford and Michael]. Allen
1 Introduction 233
2 Radioligand binding assays 233
Introduction 233
Choice of reagents 234
Separation techniques 236
Validation and optimization of assay 240
Data analysis 241
Non radioactive assays 245
3 G protein coupled receptor second messenger assays 246
Introduction 246
Measurement of [35S]GTP yS binding 247
Measurement of 3 ,5 cyclic adenosine monophosphate (cAMP) 250
Measurement of inositol triphosphate 253
Non radioactive assays 255
4 Enzyme assays 256
Introduction 256
Measurement of nitric oxide synthase activity using [14C]arginine 256
Homogeneous assays 261
5 Ion channel assays 261
Introduction 261
Measurement of Na+ channel function by [14C]guanidine flux 261
6 Some commercially available curve fitting programs 263
Acknowledgements 263
References 263
10 Summary of legislation in radiological protection in the UK 265
David Prime and Barry Frith
1 Introduction 265
2 The Radioactive Substances Act (1993) 265
Acquisition 265
Storage 265
Radioactive waste 265
3 The Ionising Radiations Regulations (1999) 266
Interpretation and general 266
General principles and procedures 266
Arrangements for the management of radiation protection 268
Designated areas 268
Classification and monitoring of persons 268
Arrangements for the control of radioactive substances, articles and
equipment 269
Duties of employees and miscellaneous 269
11 Federal regulations on use of radionuclides in the USA 271
Eileen D. Hotte and Neal Nelson
1 Introduction 271
2 Licensing requirements 273
3 General domestic license 273
4 Specific licenses of broad scope 274
5 Medical uses 275
6 Exemptions 276
7 Radiation protection—Nuclear Regulatory Commission 276
Immediate telephone report 279
Twenty four hour telephone report 279
Telephone report within 30 days 280
30 day written report 280
8 Other sections of Title 10, Code of Federal Regulations 280
9 Radiation protection—Department of Labor 280
10 Transportation 281
Department of Transportation 281
United States Postal Service 281
11 Environment—Environmental Protection Agency 282
12 Human uses—Department of Health and Human Services 282
13 Historical perspective and future developments 283
14 Conclusion 287
References 287
Appendices 289
Al Properties and radiation protection data of isotopes commonly used in the
biological sciences 290
A2 Types of monitor 291
A3 Curie becquerel conversion chart 293
A4 Calculation of amount of radioactivity remaining (RR) after a given half life
(NJ has passed 294
A5 Units commonly used to describe radioactivity 295
A6 Licensing and advisory authorities 296
A7 List of suppliers 297
Index 303
Protocol list
Suggested methods for making risk assessments
Example of a risk assessment form for a research project 42
An example of a Special Risk Assessment form 44
Hazards and risks to health from the use of radioactive substances and ionizing
radiation
Example of a form of controlling the ordering of radioisotopes, and the risk assessment
for their use 47
Use of X ray intensifying screens to increase sensitivity for 32P and ? ray emitters
Procedure for indirect autoradiography using intensifying screen 69
Fluorography of weak p emitters
Technical procedures for fluorography of acrylamide gels 71
Procedure for impregnating acrylamide gels with PPO using dimethyl sulphoxide 74
Procedure for impregnating acrylamide gels with PPO using acetic acid 75
Procedure for impregnating acrylamide gels with sodium salicylate 75
Fluorography of agarose gels 76
Fluorography of paper chromatograms 77
Fluorography of thin layer chromatograms 78
Fluorography of nitrocellulose 78
The liquid scintillation counter
Preparation of quench standards 106
D.p.m. correction without quench curves
Efficiency extrapolation calibration and sample counting 110
Sample preparation
Preparation of general purpose organic cocktail 116
Preparation of tissue samples 117
Liquid scintillation counting of plasma or serum 118
Gamma counters
Setting counting windows 122
Determination of a gamma counter s resolution 123
Storage phosphor technology
General procedure for obtaining phosphor image 127
Labelling methods for hybridization probes
DE81 chromatography 135
Random prime labelling 137
Nick translation 139
Transcription labelling 142
3 End labelling 146
End repair using Klenow polymerase 148
5 End labelling ISO
Cycle sequencing with 33P labelled terminators
Cycle sequencing with 33P labelled terminators 155
Labelling of proteins with 125I
Radio iodination using chloramine T 157
Direct radio iodination using Iodo Gen™ 158
Radio iodination using Iodo Beads™ 159
Radio iodination using lactoperoxidase 160
Conjugation with (125I) Bolton Hunter reagent 162
Labelling of proteins with 3H and C
3H LabellingwithN succinimidyl(2,3 3H)propionate 164
14C Labelling by reductive methylation 165
Protein kinase assays
Filter method for kinase assay 166
In vitro translation with radiolabelled amino acids
In vitro translation 169
Subcellular localization of biological molecules
General scheme for separation of subcellular fractions from brain using density
gradient centrifugation 172
Subcellular localization of proteins
Protein separation using polyacrylamide gels 173
Electroblotting 174
Procedure for staining membrane replicas 176
Protocol for immunoprecipitation 177
Light microscopic radioimmunocytochemistry using biotinylated protein A and
(strept)avidin [3H]biotin complex 180
Subcellular localization of nucleic acids
Preparation of frozen tissue blocks for in situ hybridization 187
Preparation of cultured cells for in situ hybridization 188
Procedure for siliconizing glass cover slips 189
Alkaline hydrolysis of RNA probes 191
Pre treatment of tissues before hybridization 193
Procedure for pre hybridization and hybridization 194
Procedure for stringency washes 195
Procedure for aliquoting new emulsion 196
Procedure for dipping slides in nuclear track emulsion 196
Procedure for developing autoradiographs 197
A Procedure for counterstaining with Pyronin Y 198
Denaturing gel electrophoresis of RNA 201
Procedure for de ionizing formamide 202
Capillary transfer of RNA to a membrane support 202
Fixing RNA to nylon membrane 203
Hybridization of northern blot 204
Radioimmunoassays
Determination of an antibody dilution curve for anti interleukin 1 216
Sample pre treatment for the removal of insulin like growth factor 1 (IGF 1) binding
proteins 218
Extraction of growth hormone releasing factor from serum 218
Radioimmunoassay for IGF 1 220
Radioimmunoassay for growth hormone releasing factor (GRF) 221
Preparation of cellulose linked secondary antibody 221
Microtitre plate IRMA 222
Immunoradiometric assays
Bead IRMA 228
Radloligand binding assays
Preparation of crude brain membranes 234
Preparation of membranes of Chinese hamster ovary (CHO) cells 235
Measurement of the binding of 3H labelled substance P in membranes prepared from
rat cerebral cortex 237
G protein coupled receptor second messenger assays
Measurement of [35S]GTPS binding in membranes prepared from Chinese hamster
ovary (CHO) cells stably expressing human melatonin mt, receptors 247
Preparationof CHO cells stably transfected with human p adrenoceptor and extraction
of cAMP for measurement of cAMP by 125I Scinitillation Proximity Assay 250
[i25j] cAMP SPA of cAMP from samples generated in protocol 5 251
Measurement of total inositol phosphate accumulation in CHO cells transfected with
human oxytocin receptor 253
Enzyme Assays
Extraction of nitric oxide synthase from tissue or cell extracts 257
Measurement of nitric oxide synthase activity using [ 14C ] arginine 258
Ion channel assays
Measurement of [14C]guanidine flux into CHO cells stably expressing recombinant
Na+ channels 262
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| geographic | Großbritannien USA |
| geographic_facet | Großbritannien USA |
| id | DE-604.BV013838002 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T18:52:56Z |
| institution | BVB |
| isbn | 0199638276 0199638268 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009464991 |
| oclc_num | 47521283 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-634 |
| owner_facet | DE-355 DE-BY-UBR DE-634 |
| physical | XX, 307 S. Ill., graph. Darst. |
| publishDate | 2002 |
| publishDateSearch | 2002 |
| publishDateSort | 2002 |
| publisher | Oxford Univ. Press |
| record_format | marc |
| series | The practical approach series |
| series2 | The practical approach series |
| spelling | Radioisotopes in biology a practical approach ed. by R. J. Slater 2. ed. Oxford [u.a.] Oxford Univ. Press 2002 XX, 307 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series 252 Biologia (estudos específicos) larpcal Imunoensaio larpcal Traçadores radioativos larpcal Autoradiography methods Great Britain Autoradiography methods United States Radioactive tracers in biology Laboratory manuals Radioisotopes Great Britain Radioisotopes United States Safety legislation & jurisprudence Great Britain Safety legislation & jurisprudence United States Scintillation Counting methods Great Britain Scintillation Counting methods United States Isotopenmarkierung (DE-588)4176811-5 gnd rswk-swf Tracer (DE-588)4185795-1 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Radionuklid (DE-588)4133541-7 gnd rswk-swf Biologie (DE-588)4006851-1 gnd rswk-swf Großbritannien USA Radionuklid (DE-588)4133541-7 s Biologie (DE-588)4006851-1 s Methode (DE-588)4038971-6 s 1\p DE-604 Isotopenmarkierung (DE-588)4176811-5 s 2\p DE-604 Tracer (DE-588)4185795-1 s 3\p DE-604 Slater, Robert J. Sonstige oth The practical approach series 252 (DE-604)BV007921321 252 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009464991&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 2\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 3\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Radioisotopes in biology a practical approach The practical approach series Biologia (estudos específicos) larpcal Imunoensaio larpcal Traçadores radioativos larpcal Autoradiography methods Great Britain Autoradiography methods United States Radioactive tracers in biology Laboratory manuals Radioisotopes Great Britain Radioisotopes United States Safety legislation & jurisprudence Great Britain Safety legislation & jurisprudence United States Scintillation Counting methods Great Britain Scintillation Counting methods United States Isotopenmarkierung (DE-588)4176811-5 gnd Tracer (DE-588)4185795-1 gnd Methode (DE-588)4038971-6 gnd Radionuklid (DE-588)4133541-7 gnd Biologie (DE-588)4006851-1 gnd |
| subject_GND | (DE-588)4176811-5 (DE-588)4185795-1 (DE-588)4038971-6 (DE-588)4133541-7 (DE-588)4006851-1 |
| title | Radioisotopes in biology a practical approach |
| title_auth | Radioisotopes in biology a practical approach |
| title_exact_search | Radioisotopes in biology a practical approach |
| title_full | Radioisotopes in biology a practical approach ed. by R. J. Slater |
| title_fullStr | Radioisotopes in biology a practical approach ed. by R. J. Slater |
| title_full_unstemmed | Radioisotopes in biology a practical approach ed. by R. J. Slater |
| title_short | Radioisotopes in biology |
| title_sort | radioisotopes in biology a practical approach |
| title_sub | a practical approach |
| topic | Biologia (estudos específicos) larpcal Imunoensaio larpcal Traçadores radioativos larpcal Autoradiography methods Great Britain Autoradiography methods United States Radioactive tracers in biology Laboratory manuals Radioisotopes Great Britain Radioisotopes United States Safety legislation & jurisprudence Great Britain Safety legislation & jurisprudence United States Scintillation Counting methods Great Britain Scintillation Counting methods United States Isotopenmarkierung (DE-588)4176811-5 gnd Tracer (DE-588)4185795-1 gnd Methode (DE-588)4038971-6 gnd Radionuklid (DE-588)4133541-7 gnd Biologie (DE-588)4006851-1 gnd |
| topic_facet | Biologia (estudos específicos) Imunoensaio Traçadores radioativos Autoradiography methods Great Britain Autoradiography methods United States Radioactive tracers in biology Laboratory manuals Radioisotopes Great Britain Radioisotopes United States Safety legislation & jurisprudence Great Britain Safety legislation & jurisprudence United States Scintillation Counting methods Great Britain Scintillation Counting methods United States Isotopenmarkierung Tracer Methode Radionuklid Biologie Großbritannien USA |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009464991&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV007921321 |
| work_keys_str_mv | AT slaterrobertj radioisotopesinbiologyapracticalapproach |