Jan 29, 2024
Labeling lysosomes in the adult Drosophila brain using LysoTracker
- Mel Feany1,2
- 1Department of Pathology, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115;
- 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815
Protocol Citation: Mel Feany 2024. Labeling lysosomes in the adult Drosophila brain using LysoTracker. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlkojjwv5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: January 29, 2024
Last Modified: May 31, 2024
Protocol Integer ID: 94349
Keywords: ASAPCRN
Abstract
This protocol describes the labeling and quantification of Lysosomes in the adult Drosophila brain using the LysoTracker dye
Prepare the working solution, 0.05 μM LysoTracker Green DND26, using 1X PBS
Brains from 10-day-old animals are dissected in ice-cold 1X PBS
The dissected brain is transferred to a glass slide, and the excess 1X PBS removed
The brains are re-incubated with 0.05 μM LysoTracker for 00:05:00 at Room temperature
5m
After 5 minutes, the excess solution is removed, a drop of DAPI Fluoromount added, and the brain mounted with a coverslip
Brains re imaged immediately using a Zeiss confocal microscope under a 63X objective lens
The number of puncta per 1000 μm2 is counted in Zen software