Hello,
I am working with primary mouse cortical neurons plated in 96-well plates and I will need to image the mitochondria. I have read around and decided to use MitoTracker deep red as it can be retained after fixing cells with PFA. When I look at the protocol it advises to use concentrations between 25-500 nm (and between 100 and 500 nm if the cells are going to be fixated). I have been trying to find publications that used this method to observe the optimal concentration to use in mouse cortical neurons but unfortunately could not find anything suitable. Could someone advise me on what concentration it would be best to use in my circumstances? If you have a specific protocol that you use I would greatly appreciate if you could share it with me so I can take a look.
Hi, please see the attached links that describes in detail the protocol for staining cultured neuronal cells with mitotracker deep red. In one of the paper, authors have used a final concentration of 100 nM.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7986497/
https://onlinelibrary.wiley.com/doi/full/10.1002/cyto.a.21061
I hope this helps. Good luck,
Alpana