Hi!

I am trying to obtain good-quality 16um transversal cryosections of the lumbar region of the mouse spinal cord for subsequent Nissl staining. Please help me learn the best practices out there: starting with the best freezing method, the most suitable cryo molds, and embedding medium, and maybe some other tips and tricks.

For now, I embed in M1 and freeze fixed tissue on dry ice with ethanol in a cubicle cryomold. I section at -20 and flatten my sections using the anti-role plate technique, which I then fish out by bringing a superfrost positively charged microscope slide close to the section until it sticks to the slide itself ( i am afraid that if i will touch the specimen i will squish it, however, i noticed that this self-adhesion technique is not ideal either as it leads to the folding of the specimen).

Finally, is there a trick to identify by eye L1 to L6 regions while sectioning, before the Nissl stain?

Thank you all in advance! Wish you a wonderful day!