Sidney,
Wash your slides after anilinblue in acidified aqua dest. for 20-30 seconds.
After this go directly into 100% ethanol, with 3 changes also for 20-30
seconds, then xylol and mount.
Diluted ethanol tends to differentiate the staining too much.
The literature says, that washing in water and ethanol decreases the
intensity. I made some tests. After 30 min in acidified water or ethanol,
the colours were paler but distinct.

Gudrun Lang
 
Biomed. Analytikerin
Histolabor
Akh Linz
Krankenhausstr. 9
4020 Linz
+43(0)732/7806-6754
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Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Sydney
Huang
Gesendet: Samstag, 19. Jänner 2008 03:54
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Does alcohol afeects the staining results of
Masson'sTrichrome

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

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