Greetings, I have an investigator who sent the note below to me after we together did the FACS and fl. microscope observation...... The cells treated with valinomycin were nicely depolarized and showed a stong green (little orange) signal. The cells in question were bright orange and indeed full of bright orange mitochondrion. Together we are stumped...hence the note to , as we say here in Da Bronx.....Da List! > >I'm using JC-1 to look at the time couse of mitochondrial specific events >in drug-induced apoptosis. Basically, I treat my cells with drug for >various time points (Up to 24hrs maximum), after which I harvest adherent >and non-adherent cells and stain for 15 min at 37 degrees in 1uM of JC-1. >Instead of depolarisation (a change from orange aggregate formation to >green fluorescence), I see a hyperpolarization ....my drug treated cells >are bright orange (even more than controls), reaching a maximum intensity >at the 24 hr time point. These cells I know to be 80 - 90 % apoptotic, and >I can even see a classic apoptotic morphology (blebbed membranes) with the >bright orange mitochondrial staining. > >Since the drug I'm using is very hydrophobic could this be having an effect >on the mitochondrial membrane potential alone...or could it somehow be >accumulating in mitochondria? > >Any suggestions would be gratefully appreciated. > > >Hayley McDaid, Ph.D. >Molecular Pharmacology >Golding Room 201 >Albert Einstein College of Medicine >1300 Morris Park Avenue >Bronx, New York 10461 > >Telephone: 718-430-2192 >Fax: 718-430-8922 >E-mail: mcdaid@aecom.yu.edu > > > > Dave Gebhard--Director FACS Facility Albert Einstein College of Medicine Chanin Building 309 1300 Morris Park Avenue Bronx, New York 10461 718-430-2724/ 3573 FAX 718-430-8550
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