Flow Cytometry: A Practical Approach

Capa
M. G. Ormerod
OUP Oxford, 18 de mai. de 2000 - 276 páginas
Flow Cytometry 3/e is intended as a handbook for every laboratory that has a bench-top flow cytometer or a fluorescence activated cell sorter. It is an introduction and guide to those new to the field and a first point of reference for experienced practitioners who want to investigate a new technique. The chapter on immunophenotyping - the most important clinical application of flow cytometry - has been strengthened by the addition of a chapter on quality control in the clinical laboratory. The utility of the book in a clinical laboratory has been further enhanced by the addition of a chapter covering ten other clinical applications. Flow cytometry has found increasing application in the field of apoptosis research. A new chapter has been added to cover this important topic. Every flow cytometry laboratory can't afford not to have a copy on the shelf as a first point of reference. The book is not fully comprehensive but it does aim to cover over 90% of the applications of flow cytometry in mammalian biology.
 

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Conteúdo

Introduction to the principles of flow cytometry
1
2 Techniques for sample preparation
2
3 Fluidics
3
4 Detection and measurement
7
5 Cell sorting
21
References
22
Fluorescence and fluorochromes
23
3 Fluorochromes used to label covalently other probes
25
References
132
Measurement of cytoplasmic and nuclear antigens
133
2 Methods of cell permeabilization
134
3 Methods of cell staining and analysis
142
4 Examples of intracellular antigens that can be measured by flow cytometry
146
References
155
Analysis of DNAmeasurement of cell kinetics by the bromodeoxyuridineantibromodeoxyuridine method
159
2 Basic cellkinetic concepts
160

4 Fluorochromes used to label directly cell components
28
References
33
Preparing suspensions of single cells
35
3 Leucocytes from other tissue
38
4 Other types of cell
39
5 Cultured cells
44
References
45
Flow sorting
47
3 Parameters which affect sorting
50
4 Practical considerations
52
5 Specialized features
57
6 Other flowsorting systems
58
Further reading
59
Immunofluorescence of surface markers
61
2 Instrument standardization
63
3 Testing sensitivity and resolution
68
4 Immunofluorescence
70
6 Titration of antibodies
72
9 Constructing an immunophenotyping panel
73
10 Isotype controls
80
11 DNA contentsurface marker analysis
81
References
82
Analysis of DNAgeneral methods
83
3 General comments
85
4 Experimental methods
89
5 Deconvolution of the DNA histogram
93
Acknowledgement
96
Further clinical applications
99
3 The enumeration of CD34+ cells in peripheral blood stemcell harvests and bonemarrow harvests
101
4 Simultaneous detection of granulocyte and lymphocytereactive antibodies
104
5 Detection of plateletreactive antibodies
107
6 Application of crossmatching by flow cytometry to transplantation
109
7 Monitoring antithymocyte globulin ATGantilymphocyte globulin ALG therapy in transplantation
111
8 Measurement of cellmediated cytotoxicity by flow cytometry
114
9 Monitoring cell activation in response to antigens or mitogens
116
10 Measuring intracellular cytokines by flow cytometry
117
References
122
Quality assurance in the clinical laboratory
125
3 Internal qualitycontrol procedures
127
4 External quality assurance
130
5 Conclusions
131
3 Background to the BrdUrd technique
162
4 Methods of BrdUrd incorporation
164
5 Tissue preparation
166
6 Staining procedures
167
7 Flow cytometry
172
9 Conclusions
176
Acknowledgements
177
Analysis of cell proliferation using the bromodeoxyuridineHoechstethidium bromide method
179
3 Cell staining and flow cytometry
180
4 Synchronized cells
182
5 Asynchronous cells
183
6 Troubleshooting
186
Acknowledgements
188
Chromosome analysis and sorting by flow cytometry
189
2 Preparation of chromosome suspensions
192
3 Instrumentation
196
4 Flow sorting chromosomes for library construction
197
5 Generation of chromosome paints
198
References
201
Intracellular ionized calcium magnesium membrane potential and pH
203
2 Magnesium
222
4 Measurement of cytoplasmic pH
227
Acknowledgement
232
Flow cytometry in the study of apoptosis
235
3 Measurement of DNA degradation
236
4 Mitochondrial membrane potential
240
5 Measurements of changes in the plasma membrane
243
6 Other measurements
245
7 Quantification of apoptotic cells
246
Acknowledgement
247
Further applications to cell biology
249
3 Monitoring electropermeabilization of cells
251
4 Measurement of oxidative burst
253
5 Characterizing multidrug resistance MDR in cancer cells
254
Safety procedures
259
3 Chemicals
260
Flow cytometers and software
261
2 Software for data manipulation
263
List of suppliers
265
Index
273
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