I am looking for detailed protocol for examining mitochondrial membrane potential using Rh123 and PI, also for Sperm Cell Structure Assay for rams for flow cytometrical analysis.
The Rho123 is a membrane permeable cationic fluorescent dye that accumulates into mitochondria with respect to its negative trans-membrane potential
(ΔΨm). The ΔΨm can be monitored using Rho123 as described in our published paper 2016. For more detail please see journal In Vitro Cell.Dev.Biol.—Animal
As an alternative, I can suggest the use of JC-1. I published my modified method for immunofluorescence staining (Article Comparison of Cellular Death Pathways after mTHPC-mediated P...
), but it can easily ba adapted to flow cytometry (as explained in the manufacturer's manual: www.bdbiosciences.com/us/applications/research/apoptosis/apoptosis-kits-sets/bd-mitoscreen-jc-1/p/551302 ).