[Histonet] Does alcohol afeects the staining results of Masson's Trichrome

[Sydney Huang]

Hi all

I am currently encounter a problem when doing masson's trichrome, and
I hope that
someone can help me.  The last steps in trichrome stain which are to
dehydrate and clear in xylene before mount the slide, this is where it
troubles me.  After all the staining before dehydrating I checked the
color under microscope and all colors were great, collagen blue,
cytoplasma and smooth muscle red, and nucleus black.  However after
the dehydrating and clearing process the redness from the cytoplasma
was lost.  I use 2 changes of 95% alcohol then 2 changes of 100%
alcohol 1 minutes each, then 2 changes of xylene also 1 minutes each.
Does the freshness of the alcohol affects the results?  If so, how
often should I change the alcohol?

Thanks in advance

sincerely yours
Sydney