Labeling lysosomes in the adult Drosophila brain using LysoTracker
Mel Feany
Published: 2024-01-30 DOI: 10.17504/protocols.io.j8nlkojjwv5r/v1
Abstract
This protocol describes the labeling and quantification of Lysosomes in the adult Drosophila brain using the LysoTracker dye
Steps
1.
Prepare the working solution, 0.05 μM LysoTracker Green DND26, using 1X PBS
2.
Brains from 10-day-old animals are dissected in ice-cold 1X PBS
3.
The dissected brain is transferred to a glass slide, and the excess 1X PBS removed
4.
The brains are re-incubated with 0.05 μM LysoTracker for 0h 5m 0s at Room temperature
5.
After 5 minutes, the excess solution is removed, a drop of DAPI Fluoromount added, and the brain mounted with a coverslip
6.
Brains re imaged immediately using a Zeiss confocal microscope under a 63X objective lens
7.
The number of puncta per 1000 μm2 is counted in Zen software
