[Histonet] Masson's Trichrome Troubleshooting

[Ford, Judi]

Hi everyone,

 

I'm hoping someone can give me suggestions on what happened to my stain.
I did a Masson's Trichrome on mouse hearts/aortas this morning.  I
followed the AFIP protocol exactly and used two different control slides
(tongue, aorta, skeletal muscle). We have previously stained control
slides with the tissue I used this time and they worked beautifully.

 

After finishing the glacial acetic acid rinse I examined the slides
under the scope and found that neither of the control slides stained
with the aniline blue, but all the study slides stained beautifully with
aniline blue. I tried going back and restaining (from
phosphomolybdic/phosphotungstic acid) the slides and the same results
showed. I thought maybe I had them in the glacial acetic acid too long
so I cut that back to 3 minutes instead of 5 for the second round.

 

Any ideas on what could have happened? Could the control slides have
been sitting too long as unstained slides? Could the aniline blue
working solution need an extra punch from glacial acetic acid? In the
past I've reused aniline blue without any problems and this solution
wasn't past its expiration date.

 

Would love to hear your thoughts......

 

Judi Ford

Palo Alto, CA