Whitaker Calcium in Living Cells

2. Auflage 2010
ISBN: 978-0-08-095122-5
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: 6 - ePub Watermark

E-Book, Englisch, Band Volume 99, 328 Seiten

Reihe: Methods in Cell Biology

Every cell of the body is dependent on calcium to function. Calcium is found in teeth and bones, and calcium signalling is necessary for the movement of muscles and for the action of the heart and the intestines as well as blood coagulation. Calcium in Living Cells will update classic techniques in detecting microscopic levels of calcium ions (Ca2+) in living cells, as well as address new techniques in the field of calcium detection and calcium signaling. Such detection and measurement of intracellular calcium is important to researchers studying the heart, musculoskeletal, gastrointestinal, and immune systems, whose findings will aid in the advancement of drug and genomic therapies to treat heart, gastrointestinal, autoimmune, and infectious diseases. - Gives researchers much needed information on how to study calcium in live cells, which is becoming increasingly important in heart, musculoskeletal, and immune system research - Provides an overview of the latest methods--fluorescence resonance energy transfer (FRET), for example-- that are new to the field - Allows understanding of how calcium plays a role in intracellular function at the cellular level, which has proved important in Alzheimer's research, heart disease, and areas of musculoskeletal research - Updated chapters reflect advancements in the classic techniques used'preparing calcium buffers, vibrating the Ca2+ Electrode and confocal imaging

Autoren/Hrsg.

Whitaker, Michael

Inhaltsverzeichnis

1;Front Cover;1
2;Methods in Cell Biology;4
3;Copyright Page;5
4;Contents;6
5;Contributors;10
6;Preface;12
7;Chapter 1: A Practical Guide to the Preparation of Ca2+ Buffers;14
7.1;I. Introduction;15
7.2;II. Rationale;16
7.3;III. Methods;19
7.4;IV. Materials;25
7.5;V. Discussion and Summary;36
7.6;Acknowledgments;37
7.7;References;37
8;Chapter 2: Photorelease Techniques for Raising or Lowering Intracellular Ca2+;40
8.1;I. Introduction;41
8.2;II. Nitr Compounds;42
8.3;III. DM-Nitrophen;48
8.4;IV. Diazo Compounds;52
8.5;V. Introduction into Cells;55
8.6;VI. Light Sources;56
8.7;VII. Calibration;58
8.8;VIII. Purity and Toxicity;61
8.9;IX. Biological Applications;62
8.10;X. Conclusions;71
8.11;Acknowledgments;72
8.12;References;72
9;Chapter 3: Making and Using Calcium-Selective Mini- and Microelectrodes;80
9.1;I. Introduction;81
9.2;II. Rationale;83
9.3;III. Methods;84
9.4;IV. Discussion;99
9.5;Acknowledgments;100
9.6;References;100
10;Chapter 4: Construction, Theory, and Practical Considerations for using Self-referencing of Ca2+-Selective Microelectrodes forMonitoring Extracellular Ca2+ Gradients;104
10.1;I. Introduction;105
10.2;II. CaSM Construction;106
10.3;III. Properties of CaSMs;107
10.4;IV. Self-referencing of CaSMs;111
10.5;V. Ca2+ Flux Measurements ;120
10.6;Acknowledgments;121
10.7;References;122
11;Chapter 5: Practical Aspects of Measuring Intracellular Calcium Signals with Fluorescent Indicators;126
11.1;I. Introduction;127
11.2;II. Fluorescent Ca2+ Indicators;127
11.3;III. Loading Indicators into Cells;133
11.4;IV. Manipulation of [Ca2+];139
11.5;V. Conversion of Indicator Fluorescence Signal into Values of [Ca2+];145
11.6;VI. Reporting Indicator Fluorescence Intensity Changes without Calibration;152
11.7;VII. Measuring [Ca2+] in Mitochondria;154
11.8;VIII. Concluding Remarks;159
11.9;Appendix 1. The fraction of a polybasic acid that exists in a particular state of protonation;160
11.10;Appendix 2. Deriving an expression for the amount of fluorescence emitted by a solution of fluorescent indicator;162
11.11;References;164
12;Chapter 6: Genetically Encoded Probes for Measurement of Intracellular Calcium;166
12.1;I. Introduction;167
12.2;II. Genetically Encoded Sensors;168
12.3;III. Applications of Genetically Encoded Sensors;180
12.4;IV. Use of Genetically Encoded Calcium Sensors;189
12.5;V. Conclusions;190
12.6;Acknowledgments;190
12.7;References;190
13;Chapter 7: Patch Clamp Methods for Studying Calcium Channels;196
13.1;I. Introduction;197
13.2;II. Rationale;199
13.3;III. Methods;200
13.4;IV. Materials;208
13.5;V. Discussion;208
13.6;VI. Summary;209
13.7;Acknowledgments;209
13.8;References;209
14;Chapter 8: Nuclear Patch-Clamp Recording from Inositol 1,4,5-Trisphosphate Receptors;212
14.1;I. Introduction;213
14.2;II. Nuclear Patch-Clamp Recording;214
14.3;III. Choice of Cells for Analyses of IP3R;218
14.4;IV. Methods;219
14.5;V. Concluding Remarks;231
14.6;Acknowledgments;232
14.7;References;232
15;Chapter 9: Confocal and Multiphoton Imaging of Intracellular Ca2+;238
15.1;I. Why Study Ca2+ Signaling with Confocal and Multiphoton Microscopy;239
15.2;II. Confocal Microscopy;240
15.3;III. Limitations in Speed of Confocal Imaging;243
15.4;IV. Laser Scanning Confocal Microscopy;243
15.5;V. Total Internal Reflection Fluorescence Microscopy;246
15.6;VI. Förster Resonance Energy Transfer Microscopy;248
15.7;VII. Parallel Scanning Confocal Systems;251
15.8;VIII. Spinning Disk Confocal Microscopy;251
15.9;IX. Programmable Matrix Microscopy;252
15.10;X. Advantages and Disadvantages of Confocal Microscopy;254
15.11;XI. Multiphoton Excitation Laser Scanning Microscopy;255
15.12;XII. Ca2+ Indicators for Use in Confocal and Multiphoton Microscopy;258
15.13;XIII. Use of Dyes for Single-Photon Confocal Microscopy;261
15.14;XIV. Use of Dyes for 2P Excitation Microscopy;261
15.15;XV. Is It Worth Converting the Intracellular Fluorescence Signal to [Ca2+]?;263
15.16;XVI. Calibration of Single Wavelength Dyes;264
15.17;XVII. Estimation of Fmax Values;265
15.18;XVIII. Estimation of Fmin or the Dynamic Range of the Dye;266
15.19;XIX. Consequence of Errors in Estimation of Intrinsic and Dye Fluorescence;266
15.20;XX. Multimodal and Multiple Fluorophore Confocal and Multiphoton Microscopy;268
15.21;References;272
16;Chapter 10: The Use of Aequorins to Record and Visualize Ca2+ Dynamics: From Subcellular Microdomains to Whole Organisms;276
16.1;I. Introduction;277
16.2;II. Expression of Apoaequorin, GFP-Apoaequorin, and Other Apoaequorin-Based Spectral Variants in Cells, Tissues, and Whole Organisms;281
16.3;III. Introducing Coelenterazines into Cells, Tissues and Embryos;296
16.4;IV. Techniques for Detecting Aequorin Luminescence ;297
16.5;V. Conclusions;305
16.6;Acknowledgments;305
16.7;References;305
17;Index;314
18;Volumes in Series;320
19;Color Plates;330