[Histonet] IHC on LR White hair follicles

[Kimberly Samayoa]

Hi Histonetters,

I am new at working with resins and have been tasked to embed eyebrow hair
in LR White for IHC staining of the hair follicles.  I thermally cured the
resin at 60C for 24 hours and sectioned the blocks at 2 micron.  I dried the
slides for 1 hour at 60C and open to air overnight.

I am stumped because I am not sure how or where to start next.  The
antibodies I'm going to use I have used in paraffin sucessfully, however,
they require pretreatment (Citrate buffer, pH 5.5 in a pressure cooker).  I
also read somewhere that H2O2 can make the sections come off the slide, so
is endogenous H2O2 blocking needed?  How does the DAB work with LR White?
Does the resin need to be removed prior to staining?

Lastly, I will be using JB4 in the near future and am curious to hear from
people who use it if it is compatible for IHC.   Is there a book someone
could suggest that would be a good guide for how to work with resins?  I
appreciate any advice and thank you in advance.

-- 
Kimberly A. Samayoa
Work Direct: (650) 624-3295