How can you observe Ca++ concentration on frog's sciatic nerve in-situ by using confocal microscope?

05 April 2017 1 8K Report

I am trying to observe change in extracellular calcium ion concentration on frog's sciatic nerve in-situ. We have fluo-4 AM as a calcium indicator dye. However, for some reason, the other parts of the glass (not the nerve fiber) illuminates more than the nerve fiber. I expect that only the nerve fiber must illuminate since the dye is connected to the calcium on the fiber, but it didn't happen. As I understood, I need to greatly wash up the sample. Which solution should I use to wash?

Sefa Şimşek

Hello Christina,

Couldn't I see the extracellular calcium with fluo-4 AM? Actually I can see the structure of nerve fibers but the illumination of the nerve is too low. Since I don't know the difference between fluo-4 and fluo-4 AM, I do not understand why AM group is not suitable for us.

Thanks for the response

Best wishes,

Sefa

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