Re: [Histonet] aldehyde fuchsin prep and double stains for specials
The basic fuchsine must be pararosaniline (CI
42500, Basic red 9). Other components of basic
fuchsine (rosaniline, new fuchsine etc) won't
work. You can get pararosaniline as a certified
dye.
The paraldehyde has to depolymerize. Hydrochloric
acid is for the acid-catalyzed hydrolysis of
paraldehyde, to generate acetaldehyde. Possibly
old HCl might have lost some potency. It is, after
all, a dissolved gas. The active substance
released is acetaldehyde, which some people use
directly. According to Peggy Wenk [J.
Histotechnol. 19(4): 353, 1996] 2.5 ml of
acetaldehyde can be substituted for 1.5 ml of
paraldehyde).
--
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
kiernan[AT]uwo.ca
http://publish.uwo.ca/~jkiernan/
http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
"Madary, Joseph" wrote:
>
> Hello my heroes of the histonet, I am having trouble with the aldehyde fuchsin stain. Not the procedure, the reagent. I have made this stuff up more times than a headhunter whining. Twice I made it up, changed everything but the basic fuchsin each time. It will not ripen to that deep purple color, and will not work in procedure either. I checked the archives and see that one of the many masters of the microtome John Kiernan recommends heating during the prep, I will try that. The paraldehyde was fresh both times, but was in the fridge at 4 degrees and froze because its melting point is like 11 degree or something. I called Sigma and they said it changes states all of the time in shipping and at the warehouse and freeze thaw on that stuff is insignificant. I never had a problem with that before anyway. Anyway, I need to do a Mast Cell stain and although the investigator is accepting TOL BLU she wants some different procedures, so I will do Torens, as well. But here is the
> other thing she wants a doubel stain for muci and mast cells, any ideas? I have tried some but will not share I want fresh thoughts.
>
> Nick Madary HT/HTL(ASCP)QIHC
> Histology Mgr, Medimmune Inc
> One Medimmune Way
> Gaithersburg, MD 20878
>
> 3013984745/6113 fax 9745
>
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