Hi Andrielle,

I have seen traces of mitochondrial membrane potential showing ATP supply by using JC-1 staining in different mammalian species. Moreover, I have used Mito Tracker Deep Red 633 as well, and the results were more correlated to sperm viability. That´s why Mito Tracker Deep Red 633 was more related to the overall mitochondrial mass than the high or low mitochondiral membrane potential. However, after using JC-1 staining (assessed simultaneously with other different fluorescent stainings such as Ethidium Homodimer to evaluate sperm membrane integrity) by using Flow Cytometry I have showed sperm marked as dead, with high mitochondrial membrane potential in midpiece. From my experience, I can say that during the apoptosis process, sperm head membrane can be damaged (e.g. because of ROS), however, there is a small sperm population that is still active (motile), due to the residual mitochondrial activity. That´s why it is possible to have dead spermatozoa (sperm head membrane damaged) with residual motility due to residual mitochondrial activity. After a while they stop. If you want to try other stainings, you could use Rhodamine 1 2 3 or other kind of Mito Tracker (they are many in the market). Hope that helps!

 Hi, 

I agree with Manuel and I sugest to  treat the spermatozoa with inositol 2 mg/ml to restore the mitocondrial membrane  potential.

see also:

Eur Rev Med Pharmacol Sci. 2010 Oct;14(10):891-6.

Inositol activity in oligoasthenoteratospermia--an in vitro study.

Colone M1, Marelli G, Unfer V, Bozzuto G, Molinari A, Stringaro A.

BR

Vittorio