Hi folks,
I need to find a stain/tracer to label the neurons within the recording sites. I was thinking to use DiI, but I am not sure that it will be helpful cause it requires a longer time in fixed tissue. As I am not interested in only detecting the electrode location, coating the shanks directly will not help me either. Any help is appreciated! TIA!
Hi Aysegul Gungor Aydin ,
Just for clarification, how are you recording your neurons? Is it extracellular recording? Can you provide some more details?
Hi Andre Dagostin ! Thanks for your reply! Yes, I'll do extracellular recordings to determine the primary auditory cortex. Then I'd like to label the neurons within the placement of the microelectrode following the recording.
Dear Aysegul,
if you are doing single-cell recordings, you may consider an in vivo electroporation approach with biocytin (https://www.jove.com/t/51359/juxtasomal-biocytin-labeling-to-study-structure-function-relationship).
If, instead, you are using multielectrodes, you could inject a virus to label your neurons and record a couple of weeks later.
Or you can try immunolabeling (see https://www.abcam.com/neuroscience/mature-neurons-markers-and-their-functions for ideas).
I suppose it depends on your research question(s) which approach and/or marker will be most suitable.
I hope this helps you a bit. Good luck!
Dear Aysegul Gungor Aydin ,
Since you mentioned you tried DiI, I imagine you're trying to deliver the fluorophore with a needle/cannula to a specific region. In that case, Martijn Sierksma idea for viruses is a really nice bet. Another strategy I can think of is to use AM ester dyes. The AM probes I used so far were for calcium imaging (oregon green or Indo), but it looks like there are some that have potential to help you achieve what you're looking for:
https://www.nature.com/articles/s41467-017-01503-6
https://www.bioscience.co.uk/userfiles/pdf/Cellular-Stains-brochure.pdf
Keep in mind they're gonna label every cell membrane though. So IDK how good they will be when identifying neurons and glial cells in your prep.
That's where the viruses would come in very handy, since they're specific to cell types.
Good luck!
André
Thanks for the great answers and very helpful! I am using mutielectrodes for recording. Using the virus would be perfect if I had enough time for its expression. However, as I have a certain behavioral timeline, so the virus is not an option for me! I will check AM ester dye!
Thanks!
A
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