Viral Infection of Intact Isolated Human or Mouse Pancreatic Islets
2:19
Reagent Preparation and Experimental Setup
5:23
Confocal Imaging
7:14
Results: Insulin Granule Exocytosis Imaging Using Neuropeptide Y-pHIuorin
9:09
Conclusion
Transcripción
The overall goal of this assay is to visualize insulin granule exocytosis in real time in intact pancreatic islets by confocal microscopy. This method will answer key questions in the diabetes field. For example, what regulates insulin exocytosis?
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We describe a protocol for visualization of insulin exocytosis in intact islets using pHluorin, a pH-sensitive green fluorescent protein. Isolated islets are infected with adenovirus encoding pHluorin coupled to the vesicle cargo neuropeptide Y. This allows for the detection of insulin granule fusion events by confocal microscopy.