[Histonet] PicroSirius Red in Frozen Sections

[Laura Avogaro]

Dear Yoanna I performed Picrosirius Red staining on my tissue frozen
sections with good results so I suggest you operate as follows:

1)Fix slide in 4% PFA (5 min);
2)Rinse in ddH2O;
3)Incubate in Phosphomolybdic Acid (2 minutes);
4)Rinse in ddH2O;
5)Incubate in Picrosirius Red for (1 h);
6)Wash in acidified water (2X, 1 min each);
7)Rinse in 70% EtOH (30”);
8)Dehydrate through ascending grades of alchol(80%, 90%, 100%);
9)Wash on Xilene (2X);
10)Mounting.

Good luck and let me know!

Bye

Laura



> Hello!
>
> I did a Picrosirius red stain using the polysciences kit in paraffin
section cut at 4microns. The stain looks great. Later on my boss asked
me
> to do the picro using the same kit but in frozen section. The stain
looks
> very, very intense. Sections are really red. When looking under the
brightfield microscope there is not yellow, the entire tissue is red.
Under the polarized microscope, we can see the fiber but no  like the
paraffin sections. Does anybody have come across this situation before?
Does the picrosirius stain only works for paraffin sections? I will
appreciate any feedback on the matter. I have been searching online for
a
> protocol of Picrosirius stain for frozens, but no luck.
>
> Thank you in advance,
>
>
>
> Yoanna Bello
>
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Laura Avogaro
University of Trento
Via delle Regole, 101 38123 Mattarello (TN) – Italy
Tel:  +39 0461 283425