RE: H&E staining
of all, what exactly are you trying to do? Is this for routine H&E staining
on paraffin processed tissues? By the looks of what you have written, and it it
sounds very confusing, you should use only one type of haematoxylin at once.
They each have different purposes and you are wasting valuable reagents trying
to mix the properties/mechanisms of the two.
Secondly; I am unsure what "Clear Rite" is but if you are using
regressive haematoxylin, then a differentiator of 0.5% or 1% acid alcohol
(depending on how fresh your stain is) in one pot is usually sufficient to
differentiate Harris' haematoxylin staining of, say, ten minutes.
Thirdly; I assume the ammonium hydroxide is the 'blueing' agent and I use
Scott's Tap Water Substitute in one pot and find that sufficient to blue
perfectly well. All of the solutions I have mentioned can be found in Bancroft
and Gamble (2002), Churchill Livingstone.
your reagents maybe try:
regressive haematoxylin in as many pots as necessary for an overall time
of 10 mins followed by wash, differentiator, wash, blueing agent, wash (all one
pot each) and then your eosin. Of course you may then have to 'tweak' this to
get the exact staining you want/prefer.
Fourthly; if you want good constructive replies in response to your pleas
for help, put a subject in the subject line. Most days I ignore anything without
a subject but today the students are finished, the sun is shinning and I am
having a good day!
Dept of Pathology,
Medicine and Health Sciences,
Sultan Qaboos University,
From: kevin williams
Sent: Sunday, June 29, 2003 1:09
I have tried out a system of a(first three) regressive Gills
Heaematoxylin, followed by a Progressive Haematoxylin, of the same
strenght and the slides are exposed to it for the same amont of time. The
regressive is constantly dilueted by water and alcohol and the regressive is
constantly exposted to the progressive Haematoxylin, due to the fact that I
use a lini stainer.
Can you give me any insits into the good quality staining I have found
using no differentiator(previously using Clear Rite) followed by Amonium
Hydroxide. The eosin then follows, only gets 10 seconds. So the best I have
found is without the Clear Rite and the Amonium Hydroxide. So please tell me
what is going on and why. Why is most important.
Thanks for all your help,
A. Kevin Willimas.
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