Check the pH of the Biebrich Scarlet. It should be around 2,5.
Prolong the PMA/PTA-step, that enhances the cytoplasma-staining and
decolorize the collagen-fibers.
How long was the tissue fixed? What fixative? - Fixing the deparaffinized
slides 1-2 hours in Bouin at 60°C, or overnight at roomtemperature should
emprove the staining. Very long (months) fixation in formaldehyd could cause
a weak cytoplasma staining.

Gudrun Lang
 
Biomed. Analytikerin
Histolabor
Akh Linz
Krankenhausstr. 9
4020 Linz
+43(0)732/7806-6754
-----Ursprüngliche Nachricht-----
Von: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Sydney
Huang
Gesendet: Montag, 03. Dezember 2007 05:31
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] Trouble in staining the cytoplasma using
Masson'strichrome

Hi everyone,

I am a student in Taiwan and currently studying the effects of
nanoparticles on mice.
Paraffin embedded lung section of mouse is used for the stain.
I use the commercial Masson's trichrome kit (H15) from Sigma-Aldrich
for the stain and followed the manufacturer's instruction.  But the
problem is that the cytoplasma showed little or no staining at all.
So I tried to extend the time of Biebrich Scarlet upt to 30min but it
showed no improvement.

Can someone give me a hint what might happened?
If the chemicals are just expired (may) will it affect the results?

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