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A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae

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10:27 min

July 18th, 2011

July 18th, 2011

25,256 Views

0:05

Title

1:33

Preparation of Yeast Strains

3:00

Cell Growth and Induction of Mitophagy by Nitrogen Starvation

4:48

Labeling of the Vacuole with CMAC-Arg

6:26

Mounting Live Yeast Cells for Fluorescence Microscopy

7:52

Visualizing Mitophagy by Fluorescence Microscopy

10:02

Conclusion

A robust approach to monitor the delivery of organelles to the acidic lumen of the yeast vacuole for degradation and recycling is described. The method relies on the specific labeling of target organelles with a genetically encoded dual-emission fluorescence pH-biosensor, and visualization of individual cells using fluorescence microscopy.

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