May 28, 2025
LysoTracker and MitoTracker Staining
- 1Icahn School of Medicine at Mount Sinai
Protocol Citation: Camille Goldman 2025. LysoTracker and MitoTracker Staining. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvow6y7l4o/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: May 15, 2025
Last Modified: May 28, 2025
Protocol Integer ID: 218388
Keywords: ASAPCRN
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-024297
NASA
Grant ID: 80ARC022CA004
NIH/NINDS
Grant ID: R01NS114239
NIH/NINDS
Grant ID: UH3NS115064
NIH/NIA
Grant ID: T32AG04968
NIH/NINDS
Grant ID: F31NS13090
New York State Department of Health
Grant ID: NYSTEM-C32561GG
Abstract
Protocol for staining live cells with commercially available dyes
MitoTracker Staining
MitoTracker Staining
Prepare fresh Astrocyte Media (AM ScienCell 1801) with 100nM of MitoTracker DeepRed (Thermo Scientific M22425)
Replace media on cells with media containing MitoTracker
Incubate for 30 min at 37°C
During incubation, prepare fresh AM media with Hochst 33342 (Thermo Scientific 62249)
Replace the MitoTracker media with Hoechst 33342 media
Incubate for 10 min at 37°C
Wash 1x with PBS
Add fresh AM media and image cells
LysoTracker Staining
LysoTracker Staining
Prepare fresh Astrocyte Media (AM ScienCell 1801) with 100nM of LysoTracker DeepRed (Thermo Scientific L12492) and Hoechst 33342 (Thermo Scientific 62249)
Replace media on cells with media containing LysoTracker and Hoechst 33342
Incubate for 10 min at 37°C
Wash 1x with PBS
Add fresh AM media and image cells