I am performing patch-clamp recordings of cells from red nucleus in slice preparations. I tried both horizontal and coronal slices but I still encounter troubles with optical visualization. I can barely see cells since the huge amount of fibers passing through the red nucleus. The thickness of slices is 200-250 um. Do you have any suggestions to get a better visualization of cells?
My dear, not really (difficult, nowadays, to catch the post-doc dealing with similar issue in late 80s!). Try to speak, in case, with physiologists of the Scarnati's team (Capozzo, Florio), involved also recently into brainstem recordings. Another leader, I believe, is K Takakusaki (albeit mostly in vivo...)
Hello Graziella,
I include a publication in which they performed patch clamp recordings from red nucleus neurons in guinea-pig slice preparations. Hope the information included in this publication is helpful and good luck!!!
M Diaz-Rios
Thanks a lot for your help! I'll let you know how things are going!!!
The only suggestions that will really help with this challenging task would be (1) use the youngest animals you can possibly get away with, and (2) cut the slices even thinner if you can still do it and keep the large cells intact.
I have previously made acute slices as thin as 150 microns on the Compresstome VF-200, although they are very delicate to transfer without folding over or getting damaged. Thinner slices helped me with myelinated areas like interpeduncular nucleus, VTA, dorsal raphe, and deep cerebellar nucleus.
Lastly, look for any tricks people are using for spinal cord slice. If it works there it is likely to help you out too.
Good luck!
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