Nov 22, 2016
Version 4
Staining protocol for unicellular protists: Mitotracker and Dapi V.4
- Protist Research to Optimize Tools in Genetics (PROT-G)
- Multicellgenomelab
Protocol Citation: Maria Rubio-Brotons: Staining protocol for unicellular protists: Mitotracker and Dapi. protocols.io https://dx.doi.org/10.17504/protocols.io.gg5bty6
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
Created: November 22, 2016
Last Modified: March 27, 2018
Protocol Integer ID: 4349
Keywords: unicellular protist, protocol for unicellular protist, mitotracker, staining protocol
Troubleshooting
2% PFA Fixation: Add PFA drop by drop and mixing carefully into 1mL cells for a final concentration of 2%PFA
Incubate cells for 4 min at room temperature
00:04:00
Centrifuge at 1000g for 5 minutes
00:05:00
Wash
Wash cells with 1XPBS (1/2)
Wash cells with 1XPBS (2/2)
Resuspend cells with 1mL 1xPBS
Add 1mM mitotracker dye.
Room temperature, protect from light
Incubate for 10 minutes.
Room temperature, protect from light
00:10:00
Add 1/1000 DAPI dye.
Room temperature, protect from light
Incubate for 5 minutes.
Room temperature, protect from light
00:05:00
Note
Total incubation times:
Mytotracker: 15min
DAPI: 5min
Wash
Wash with 1xPBS (1/2)
Wash with 1xPBS (2/2)
Centrifuge at 1000g for 5 minutes and resuspend with 1xPBS
00:05:00
Mount your slide and ready for observation!