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Methods for Imaging Intracellular pH of the Follicle Stem Cell Lineage in Live Drosophila Ovarian Tissue
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Chapters
- 00:05Title
- 00:40Disection
- 02:17Mounting
- 03:15Live Imaging
- 04:13Measuring Fluorescence Intensities in FSCs, pFCs, and Follicle
- 05:37Obtaining Fluorescence Intensity Ratios
- 06:21Results: Images of Intracellular pH
- 07:26Conclusion
We provide a protocol for imaging intracellular pH of an epithelial stem cell lineage in live Drosophila ovarian tissue. We describe methods to generate transgenic flies expressing a pH biosensor, mCherry::pHluorin, image the biosensor using quantitative fluorescence imaging, generate standard curves, and convert fluorescence intensity values to pH values.
