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Radioisotopes in biology : a practical approach / edited by R. J. Slater. — 2nd ed. — Oxford : Oxford University Press, c2002.—(58.10532/R129/2nd ed.) |
Contents
Contents
1. Introduction 1
1 The decision to use a radioisotope 1
2 Laboratory facilities 1
3 The choice of a labelled compound 1
4 Storage and purity 4
5 Experimental design 5
2. Radioisotope use 7
1 Introduction 7
2 Radioactivity 7
3 Biological effects of radiation and the basis of legislation 14
4 Biological basis of dose limits 16
5 Working with unsealed sources 18
6 Designation of'classified workers' 20
7 Controlling the risks to radiation workers 20
8 The tracer study 21
9 Personal protection in the radioisotope laboratory 24
10 Routine monitoring of area 25
11 Transport of radioactive material 26
12 Disposal of radioactive waste 28
3. Risk assessments for the use of radioisotopes in biology 31
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
5 Suggested methods for making risk assessments 37
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
8 Special hazard and risk assessment for DNA labelling/sequencing work with 32p 50
9 Risk assessments for gamma-emitting radionuclides 55
10 Risk assessments for environmental protection 5711 Risks to the health of biological research workers in relation to other risks 58
4. Radioisotope detection usingX-ray film 63
1 Introduction 63
2 Choosing a class of detection method: autoradiography versus fluorography or intensifying screens 64
3 Direct autoradiography 66
4 Use of X-ray intensifying screens to increase sensitivity for 32p and y-ray emitters 67
5 Fluorography of weak [3 emitters 70
6 General technical considerations for radioisotope detection by X-ray film Choice of film and screens 79
7 The underlying principles of radioisotope detection by X-ray film 82
5. The scintillation counter 85
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
7 Advanced topics 107
8 Special counting situations 111
9 Solid scintillators and programmable coincidence gates 113
10 Sample preparation 114
11 Gamma counters 120
12 Flow counters 124
13 Multi-detector liquid scintillation and gamma counters 125
14 Storage phosphor technology 125
6. In vitro labelling of nucleic acids and proteins 131
1 Introduction 131
2 Applications of nucleic acid labelling 133
3 Labelling methods for hybridization probes 134
4 Cycle sequencing with 3ap-labelled terminators 154
5 Applications of labelled proteins 156
6 Labelling of proteins with 125I 156
7 Labelling of proteins with 3H and 14C 164
8 Protein kinase assays 165
9 In vitro translation with radiolabelled amino acids 168
7. Subcellular localization of biological molecules 171
1 Introduction 171
2 Subcellular localization of proteins 172
3 Subcellular localization of nucleic acids 181
4 Conclusion 205
8. Radioisotopes and immunoassay 209
1 Introduction 209
2 Immunoassays: theoretical considerations 210
3 Radioimmunoassays 213
4 Immunoradiometric assays 226
5 Non-radioisotopic immunoassays 229
9. Pharmacological techniques 233
1 Introduction 233
2 Radioligand binding assays 233
3 G-protein-coupled receptor second messenger assays 246
4 Enzyme assays 256
5 Ion channel assays 261
6 Some commercially available curve-fitting programs 263
10. Summary of legislation in radiological protection in the UK 265
1 Introduction 265
2 The Radioactive Substances Act (1993) 265
3 The Ionising Radiations Regulations (1999) 266
11. Federal regulations on use of radionuclides in the USA 271
1 Introduction 271
2 Licensing requirements 273
3 General domestic license 273
4 Specificlicenses of broad scope 274
5 Medicaluses 275
6 Exemptions 276
7 Radiation protection--Nuclear Regulatory Commission 276
8 Other sections of Title 10， Code of Federal Regulations 280
9 Radiation protection--Department of Labor 280
10 Transportation 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
Appendices 289
A1 Properties and radiation protection data ofisotopes commonly used in the biological sciences 290
A2 Types of monitor 291
A3 Curie-becquerel conversion chart 292
A4 Calculation of amount of radioactivity remaining (RR) after a given half-life (NL) has passed 294
A5 Units commonly used to describe radioactivity 295
A6 Licensing and advisory authorities 296
A7 List of suppliers 297
Index 3O3
