I'm using Rhod-2 AM to view calcium flux inside dying axons.
During early hours of the treatment (4 and 6 hours), Rhod-2 AM stains the axons very brightly.
But when I add Rhod-2 AM to dying axons at 12th hour, the staining is still strong, but the axons are very fuzzy, somewhat appearing orange.
I don't think this is an issue with the Rhod-2 AM dye, as I dont add the dye and wait and record at different hours, I add the dye at different times to different sets of axons.
Interestingly, the axons that were added with the dye at 4 hours are still strongly fluorescent at 12 hours, even thought the axons appear degenerating heavily.
Is Rhod-2 AM mitochondrial membrane dependent? And if so, does it get trapped in the mitochondria?
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