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Confocal Microscopy to Measure Three Modes of Fusion Pore Dynamics in Adrenal Chromaffin Cells
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Capitoli
- 00:04Introduction
- 01:07Bovine Chromaffin Cell Culture
- 02:52Transfection with Electroporation
- 04:18Preparation for Patch-Clamp Recording and Confocal Imaging
- 05:41Patch-Clamp Recording and Confocal Imaging
- 09:01Confocal Imaging Data Analysis
- 10:21Results: Whole-Cell Voltage-Clamp Recordings and Visualization of Fusion Events Under Confocal Microscope
- 11:52Conclusion
This protocol describes a confocal imaging technique to detect three fusion modes in bovine adrenal chromaffin cells. These fusion modes include 1) close-fusion (also called kiss-and-run), involving fusion pore opening and closure, 2) stay-fusion, involving fusion pore opening and maintaining the opened pore, and 3) shrink-fusion, involving fused vesicle shrinkage.
