A variety of powerful techniques for monitoring and analysing events during signal transduction at the single cell level are described in this lab manual. An introductionary section on 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…mehr
A variety of powerful techniques for monitoring and analysing events during signal transduction at the single cell level are described in this lab manual. An introductionary section on 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.
Cell cummunication is currently one of the most interesting topics in cell and molecular biology research. Various techniques to record the events and changes during signal transduction at the single cell level are described in this lab manual.
Inhaltsangabe
I Handling of Cells in Single Cell Experiments.- 1 Maintaining Cells Under the Microscope.- 2 Simple Perfusion Chambers for Single Cell Measurements.- 3 Temperature Control on the Stage of the Microscope.- 4 A Modified U-Tube Drug Application System.- 5 Laser Microsurgery as a Tool in Single Cell Research.- II Ion Channel and Membrane Potential Measurements Using the Patch-Clamp Technique.- II-1 Introduction to the Measurement Technique.- 6 Significance of Ion Channels and Membrane Potential Changes in Cells.- 7 Practical Introduction to Patch Clamping by Simulation Experiments with Simple Electrical Circuits.- 8 Improved Electrophysiological Measurements by Series Resistance Compensation.- 9 Preparation of Patchable Plant Cell Protoplasts and a Procedure for the Improvement of Gigaseal Formation.- II-2 Single Channel Measurements.- 10 Recording and Analysis of ATP-Sensitive Potassium Channels in Inside-Out Patches of Rabbit Ventricular Myocytes.- 11 Mechanotransduction and Mechanosensitive Ion Channels in Osteoblasts.- 12 The Study of (Plant) Ion Channels Reconstituted in Planar Lipid Bilayers.- 13 Ion Channel Measurement on the Cell Nucleus.- 14 Monitoring Cytosolic Calcium and Membrane Potential from Cell-Attached Patch-Clamp Currents in Human T Lymphocytes.- II-3 Whole Cell Measurements.- 15 Membrane Potential and Action Potential Measurements in Whole Cell and Perforated Patch Configurations.- 16 Perforated Patch-Clamp Technique in Heart Cells.- 17 Measurement and Analysis of Different Aspects of Potassium Currents in Human Lymphocytes.- 18 Ionic Conductances in Chicken Osteoclasts.- 19 GABAA Receptor-Mediated Chloride Currents in Acutely Dissociated Hippocampal Neurons.- 20 Measurement of Whole Cell Potassium Currents in Protoplasts from Tobacco CellSuspensions.- 21 Inward Rectifying Potassium Conductance in Barley Aleurone Protoplasts.- III Fluorescence to Measure Intracellular Ions.- III-1 Introduction to the Measurement Technique.- 22 An Introduction to the Use of Fluorescent Probes in Ratiometric Fluorescence Microscopy.- 23 How To Use Video Cameras To Acquire Images.- 24 Strategies for Studying Intracellular pH Regulation.- III-2 Flow Cytometer Measurements.- 25 An Introduction to the Working Principles of the Flow Cytometer.- 26 Flow Cytometric Membrane Potential Measurements.- 27 Flow Cytometric Measurement of Cytosolic Calcium in Lymphocytes.- III-3 Microfluorescence Measurements.- 28 Cytosolic pH and Cell Movement Measurement in Dictyostelium.- 29 Analysis of Agonist-Induced Cell Recruitment in Terms of Intracellular Calcium Mobilization in a Population of Enzymatically Dispersed Pancreatic Acinar Cells.- III-4 Confocal Microscopy.- 30 Quantitative Confocal Fluorescence Measurements in Living Tissue.- 31 Cytosolic Calcium Measurements with Confocal Microscopy.
I Handling of Cells in Single Cell Experiments.- 1 Maintaining Cells Under the Microscope.- 2 Simple Perfusion Chambers for Single Cell Measurements.- 3 Temperature Control on the Stage of the Microscope.- 4 A Modified U-Tube Drug Application System.- 5 Laser Microsurgery as a Tool in Single Cell Research.- II Ion Channel and Membrane Potential Measurements Using the Patch-Clamp Technique.- II-1 Introduction to the Measurement Technique.- 6 Significance of Ion Channels and Membrane Potential Changes in Cells.- 7 Practical Introduction to Patch Clamping by Simulation Experiments with Simple Electrical Circuits.- 8 Improved Electrophysiological Measurements by Series Resistance Compensation.- 9 Preparation of Patchable Plant Cell Protoplasts and a Procedure for the Improvement of Gigaseal Formation.- II-2 Single Channel Measurements.- 10 Recording and Analysis of ATP-Sensitive Potassium Channels in Inside-Out Patches of Rabbit Ventricular Myocytes.- 11 Mechanotransduction and Mechanosensitive Ion Channels in Osteoblasts.- 12 The Study of (Plant) Ion Channels Reconstituted in Planar Lipid Bilayers.- 13 Ion Channel Measurement on the Cell Nucleus.- 14 Monitoring Cytosolic Calcium and Membrane Potential from Cell-Attached Patch-Clamp Currents in Human T Lymphocytes.- II-3 Whole Cell Measurements.- 15 Membrane Potential and Action Potential Measurements in Whole Cell and Perforated Patch Configurations.- 16 Perforated Patch-Clamp Technique in Heart Cells.- 17 Measurement and Analysis of Different Aspects of Potassium Currents in Human Lymphocytes.- 18 Ionic Conductances in Chicken Osteoclasts.- 19 GABAA Receptor-Mediated Chloride Currents in Acutely Dissociated Hippocampal Neurons.- 20 Measurement of Whole Cell Potassium Currents in Protoplasts from Tobacco CellSuspensions.- 21 Inward Rectifying Potassium Conductance in Barley Aleurone Protoplasts.- III Fluorescence to Measure Intracellular Ions.- III-1 Introduction to the Measurement Technique.- 22 An Introduction to the Use of Fluorescent Probes in Ratiometric Fluorescence Microscopy.- 23 How To Use Video Cameras To Acquire Images.- 24 Strategies for Studying Intracellular pH Regulation.- III-2 Flow Cytometer Measurements.- 25 An Introduction to the Working Principles of the Flow Cytometer.- 26 Flow Cytometric Membrane Potential Measurements.- 27 Flow Cytometric Measurement of Cytosolic Calcium in Lymphocytes.- III-3 Microfluorescence Measurements.- 28 Cytosolic pH and Cell Movement Measurement in Dictyostelium.- 29 Analysis of Agonist-Induced Cell Recruitment in Terms of Intracellular Calcium Mobilization in a Population of Enzymatically Dispersed Pancreatic Acinar Cells.- III-4 Confocal Microscopy.- 30 Quantitative Confocal Fluorescence Measurements in Living Tissue.- 31 Cytosolic Calcium Measurements with Confocal Microscopy.
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