There are currently, 106 guest(s) and 0 member(s) that are online.
You are Anonymous user. You can register for free by clicking here
Login
Don't have an account yet? You can create one. As a registered user you have some advantages like theme manager, comments configuration and post comments with your name.
Post subject: Getting rid of NBT/BCIP precipitate from wax sections
I have some paraffin sections that I've used for immunohistochemistry (using an alkaline phosphatase-conjugated secondary antibody and NBT/BCIP as a substrate), and coverslipped with an aqueous mounting medium (Fluoromount G). I would now like to de-coverslip and re-stain these sections with a different histochemical protocol (i.e. Masson's trichrome or H&E). Does anybody know if there is a way to "clear away" the NBT/BCIP precipitate from my previous IHC?
Any advice would be greatly appreciated.
Thanks,
Andrew
Fri Jan 06, 2012 3:55 pm
Carl
Rank: Member
Joined: Oct 11, 2004
Posts: 1871
Post subject: Hi Andrew
Don't bother to remove!
Just do your H&E over the BCIP/NBT.
Sure, if this is not appropriate for your needs/does not work very well.....
I have not tried to remove this pigment but I have sometimes noted that when I have dehydrated only SOME NISH sections ( probe detected using NBT/BCIP) the pigment would dissolve out in dehydrating alcohol) so, have 3 suggestions that I list in increasing harshness...( eg: use a Coplin jar)
1. Place in 100% alcohol
2. Place in 1%HCL in 70% alcohol ( Haemalum differentiating solution)
3. Place in 1% household bleach.
Agitate gently and rinse in tapwater after an initial say, 30secs, and examine microscopically to see if there is any effect. If, after 5mins , there is no difference in 1, move to 2....etc.
Please let us know which, if any, works and if subsequent tinctorial staining is successful.
Good luck...
Carl
You cannot post new topics in this forum You cannot reply to topics in this forum You cannot edit your posts in this forum You cannot delete your posts in this forum You cannot vote in polls in this forum
