Ebook: Signal Transduction — Single Cell Techniques

A variety of powerful techniques to monitor and analyse the events during signal transduction at the single cell level are described in this lab manual. An introductionary section for cell handling includes guidelines for constructing a perfusion chamber. A main section of the book presents protocols on fluorescence techniques such as flow cytometry, microfluorescence, ion imaging and confocal microscopy. The electrophysiological section illustrates multiple applications of the patch-clamp technique in various cell types from both animals and plants. Emphasis is put on calibration and validation of the different techniques to measure changes of membrane potential, and intracellular ion concentration or pH.

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A variety of powerful techniques to monitor and analyse the events during signal transduction at the single cell level are described in this lab manual. An introductionary section for cell handling includes guidelines for constructing a perfusion chamber. A main section of the book presents protocols on fluorescence techniques such as flow cytometry, microfluorescence, ion imaging and confocal microscopy. The electrophysiological section illustrates multiple applications of the patch-clamp technique in various cell types from both animals and plants. Emphasis is put on calibration and validation of the different techniques to measure changes of membrane potential, and intracellular ion concentration or pH.

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A variety of powerful techniques to monitor and analyse the events during signal transduction at the single cell level are described in this lab manual. An introductionary section for cell handling includes guidelines for constructing a perfusion chamber. A main section of the book presents protocols on fluorescence techniques such as flow cytometry, microfluorescence, ion imaging and confocal microscopy. The electrophysiological section illustrates multiple applications of the patch-clamp technique in various cell types from both animals and plants. Emphasis is put on calibration and validation of the different techniques to measure changes of membrane potential, and intracellular ion concentration or pH.
Content:
Front Matter....Pages I-XXVII
Front Matter....Pages 1-1
Maintaining Cells Under the Microscope....Pages 3-8
Simple Perfusion Chambers for Single Cell Measurements....Pages 9-18
Temperature Control on the Stage of the Microscope....Pages 19-23
A Modified U-Tube Drug Application System....Pages 24-30
Laser Microsurgery as a Tool in Single Cell Research....Pages 31-40
Front Matter....Pages 41-41
Significance of Ion Channels and Membrane Potential Changes in Cells....Pages 43-51
Practical Introduction to Patch Clamping by Simulation Experiments with Simple Electrical Circuits....Pages 52-87
Improved Electrophysiological Measurements by Series Resistance Compensation....Pages 88-98
Preparation of Patchable Plant Cell Protoplasts and a Procedure for the Improvement of Gigaseal Formation....Pages 99-108
Recording and Analysis of ATP-Sensitive Potassium Channels in Inside-Out Patches of Rabbit Ventricular Myocytes....Pages 111-124
Mechanotransduction and Mechanosensitive Ion Channels in Osteoblasts....Pages 125-134
The Study of (Plant) Ion Channels Reconstituted in Planar Lipid Bilayers....Pages 135-152
Ion Channel Measurement on the Cell Nucleus....Pages 153-170
Monitoring Cytosolic Calcium and Membrane Potential from Cell-Attached Patch-Clamp Currents in Human T Lymphocytes....Pages 171-184
Membrane Potential and Action Potential Measurements in Whole Cell and Perforated Patch Configurations....Pages 187-206
Perforated Patch-Clamp Technique in Heart Cells....Pages 207-213
Measurement and Analysis of Different Aspects of Potassium Currents in Human Lymphocytes....Pages 214-235
Ionic Conductances in Chicken Osteoclasts....Pages 236-245
GABAA Receptor-Mediated Chloride Currents in Acutely Dissociated Hippocampal Neurons....Pages 246-255
Measurement of Whole Cell Potassium Currents in Protoplasts from Tobacco Cell Suspensions....Pages 256-268
Front Matter....Pages 41-41
Inward Rectifying Potassium Conductance in Barley Aleurone Protoplasts....Pages 269-281
Front Matter....Pages 283-283
An Introduction to the Use of Fluorescent Probes in Ratiometric Fluorescence Microscopy....Pages 285-305
How To Use Video Cameras To Acquire Images....Pages 306-317
Strategies for Studying Intracellular pH Regulation....Pages 318-339
Flow Cytometric Measurement of Cytosolic Calcium in Lymphocytes....Pages 358-372
An Introduction to the Working Principles of the Flow Cytometer....Pages 343-347
Flow Cytometric Membrane Potential Measurements....Pages 348-357
Flow Cytometric Measurement of Cytosolic Calcium in Lymphocytes....Pages 358-372
Analysis of Agonist-Induced Cell Recruitment in Terms of Intracellular Calcium Mobilization in a Population of Enzymatically Dispersed Pancreatic Acinar Cells....Pages 375-397
Quantitative Confocal Fluorescence Measurements in Living Tissue....Pages 398-406
Cytosolic Calcium Measurements with Confocal Microscopy....Pages 409-441
Back Matter....Pages 442-457
....Pages 459-468