Hi

Also, you can see this book:

Article Theory and practice of histological techniques / edited by J...

the best way ti stain them is osmiium tetroxyde which stains black axons and myelin, the only problem is a neurotoxin so it is very dangerous to use even at a 2 p/cent concentration. if you decide to use it please read the health and safety warnings very very carefully

Hi all! 

Thank you for your suggestions. As for the Osmium Tetroxide method, it seems this step is done after fixation and before paraffin embedding. The samples I have already been embedded in paraffin and sliced and mounted on slides. Will the osmium tetroxide still work if I incubate the already-made slides in it? 

Hi, i have never done it in this way, but as far as i know you should be able to do so, osmium will attach to fatty tissue, this will not change after paraffin treatment, my protocol was after fixing 10 min in osmium then wash and mount in resin, so the fixing part will work, as i told you i never did it in that way but if you have samples to spare and decide for osmium have a go and see if it works

Hello Neha,  Toluidine Blue is as other basic dyes like cresylviolett or thionine not suitable for Axon stain. This dyes stain nuclei and Nissl-substance but no fibres. If the axons are myelinatede you can combine cresyviolett with luxol-fast blue (Kluever-Barrera-stain). But the best way to stain axons in paraffin sections is to use siver-impregnations like Bielschowsky's or Himes's silverstain  for axons. The methods are discribed in J.A. Kiernan: Histological & Histochemical Methods. Good luck!

I performed such staining on semithin sections only and the results were very nice.

Regards,

Hanna Gendek-Kubiak

Hi all, thank you again for your responses, I've realized toluidine is not suitable at all and resorted back to silver staining!