Ebook: Imaging Living Cells

Improved technology for imaging living cells, specific cellular targets and organelles is having a dramatic impact on basic and applied research. By combining optical design and molecular genetics, a new series of tools is being developed and successfully applied together with classical probes. Novel labelling strategies, better software for image enhancement and analysis are now available and allow image acquisition with greater speed and precision. This lab manual, intended as for bench-top use, is suitable for both scientists and graduate students, combines an update on the most advanced imaging procedures with detailed protocols. Examples, cleverly selected from the wide repertoire of cell pyhsiology, cover different functional aspects such as distribution of multiple ions, electrical activity, exo-endocytosis, gene expression, and the cell cycle.

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Improved technology for imaging living cells, specific cellular targets and organelles is having a dramatic impact on basic and applied research. By combining optical design and molecular genetics, a new series of tools is being developed and successfully applied together with classical probes. Novel labelling strategies, better software for image enhancement and analysis are now available and allow image acquisition with greater speed and precision. This lab manual, intended as for bench-top use, is suitable for both scientists and graduate students, combines an update on the most advanced imaging procedures with detailed protocols. Examples, cleverly selected from the wide repertoire of cell pyhsiology, cover different functional aspects such as distribution of multiple ions, electrical activity, exo-endocytosis, gene expression, and the cell cycle.
Content:
Front Matter....Pages I-XXI
Front Matter....Pages 1-1
Strategies for Quantitative Digital Imaging of Biological Activity in Living Cells with Ion-Sensitive Fluorescent Probes....Pages 3-29
High Resolution 3-D Imaging of Living Cells by Image Restoration....Pages 30-50
Front Matter....Pages 51-51
Imaging Calcium in the Cytoplasm and in Organelles with Fluorescent Dyes: General Principles....Pages 53-91
Confocal Calcium Imaging....Pages 92-114
Imaging Whole Organs — Single cell [Ca2+]i measurements in the perfused liver....Pages 115-139
Simultaneous Measurement of Membrane Current and Intracellular Calcium....Pages 140-163
Quantitative Calcium Imaging in Brain Slices....Pages 164-189
Imaging of Cytosolic Sodium and pH Using SBFI and BCECF....Pages 190-214
Common Methods for Measuring Apoptotic Cell Death by Flow Cytometry....Pages 215-228
Monitoring Synaptic Exocytosis and Endocytosis by Optical Techniques....Pages 229-259
Front Matter....Pages 261-261
Targeting, Expressing and Calibrating Recombinant Aequorin....Pages 263-283
Imaging Ca2+ in Small Mammalian Cells with Recombinant Targeted Aequorin....Pages 284-298
Analysis of Regulated Gene Expression by Microinjection and Digital Luminescence Imaging of Single Living Cells....Pages 299-326
Imaging Green Fluorescent Proteins in Mammalian Cells....Pages 327-350
The New Green Fluorescent Protein Mutants and their Applications....Pages 351-361
Imaging Green Fluorescent Protein in Transgenic Plants....Pages 362-394

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Improved technology for imaging living cells, specific cellular targets and organelles is having a dramatic impact on basic and applied research. By combining optical design and molecular genetics, a new series of tools is being developed and successfully applied together with classical probes. Novel labelling strategies, better software for image enhancement and analysis are now available and allow image acquisition with greater speed and precision. This lab manual, intended as for bench-top use, is suitable for both scientists and graduate students, combines an update on the most advanced imaging procedures with detailed protocols. Examples, cleverly selected from the wide repertoire of cell pyhsiology, cover different functional aspects such as distribution of multiple ions, electrical activity, exo-endocytosis, gene expression, and the cell cycle.
Content:
Front Matter....Pages I-XXI
Front Matter....Pages 1-1
Strategies for Quantitative Digital Imaging of Biological Activity in Living Cells with Ion-Sensitive Fluorescent Probes....Pages 3-29
High Resolution 3-D Imaging of Living Cells by Image Restoration....Pages 30-50
Front Matter....Pages 51-51
Imaging Calcium in the Cytoplasm and in Organelles with Fluorescent Dyes: General Principles....Pages 53-91
Confocal Calcium Imaging....Pages 92-114
Imaging Whole Organs — Single cell [Ca2+]i measurements in the perfused liver....Pages 115-139
Simultaneous Measurement of Membrane Current and Intracellular Calcium....Pages 140-163
Quantitative Calcium Imaging in Brain Slices....Pages 164-189
Imaging of Cytosolic Sodium and pH Using SBFI and BCECF....Pages 190-214
Common Methods for Measuring Apoptotic Cell Death by Flow Cytometry....Pages 215-228
Monitoring Synaptic Exocytosis and Endocytosis by Optical Techniques....Pages 229-259
Front Matter....Pages 261-261
Targeting, Expressing and Calibrating Recombinant Aequorin....Pages 263-283
Imaging Ca2+ in Small Mammalian Cells with Recombinant Targeted Aequorin....Pages 284-298
Analysis of Regulated Gene Expression by Microinjection and Digital Luminescence Imaging of Single Living Cells....Pages 299-326
Imaging Green Fluorescent Proteins in Mammalian Cells....Pages 327-350
The New Green Fluorescent Protein Mutants and their Applications....Pages 351-361
Imaging Green Fluorescent Protein in Transgenic Plants....Pages 362-394
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