Hi all,
I am trying to image zinc in our hippocampal neurons around DIV 12 - 21. I tried both New Port Green and Fluozin-3 (from 1 - 5 uM, load 30 min - 1 hour at RT, wash for 30 min) and the neurons seem to be loaded really well. We applied several zinc solutions and imaged them live. However, when we tried to combine these dyes with neurons transfected with mcherry, it did not turn out really well. Most of the transfected neurons were not at all loaded with the dyes very nicely. Has anyone experienced this? Do you have any hints of how this should be improved?
Also, have any of you tried to fix these dyes instead of doing live imaging?
Thank you so much for your help!
Huong
Hi Micheal, I am not sure what you meant with before and after images? Do you want to see the neurons before loading and after loading? I only have images of them after loading with the dyes in green and red fluorescent channels. Would you like to see that? We just tried again yesterday and loaded them with FluoZin-3 2 uM for 1 hour at RT, this seems to help visualizing the fine processces! Previously we used FluoZin-3 at 1 uM for 30 min and that is not at all enough to see the dendrites or any finer process. :) Anyone has other trick?
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