Measuring Fluorescence Intensities in FSCs, pFCs, and Follicle
5:37
Obtaining Fluorescence Intensity Ratios
6:21
Results: Images of Intracellular pH
7:26
Conclusion
Transcript
The overall goal of this procedure is to measure the intracellular pH of follicle cells within live explanted Drosophila ovarian tissue. This method can help us answer key questions in stem cell and developmental biology such as, how changes to in
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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.