Handling of Cells in Single Cell Experiments -- Ion Channel and Membrane Potential Measurements Using the Patch-Clamp Technique: Introduction to the Measurement Technique. Single Channel Measurements. Whole Cell Measurements -- Fluorescence to Measure Intracellular Ions: Introduction to the Measurement Technique. Flow Cytometer Measurements. Microfluorescence Measurements. Confocal Microscopy -- Appendix: List of Suppliers.
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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.