Re: Histo in Plastic
<< Previous Message | Next Message >>
From: | "Karen D. Larison" <LARISONK@UONEURO.uoregon.edu> |
To: | histonet@Pathology.swmed.edu |
Reply-To: | |
Date: | Wed, 09 Jun 1999 17:05:18 -0800 |
Content-Type: | |
We got plenty of those "old" way recipes round here:
Oversight stains for plastic sections
Lee's methylene blue basic fuchsin
Stock solutions
1. 0.13 gms methylene blue in 100 ml DW
2. 0.13 gms basic fuchsin in 100 ml DW
3. 0.2 M phosphate buffer, pH 7.2 to 8.0
Staining solution: Mix
methylene blue 12 ml
basic fuchsin 12 ml
0.2 M PO4 buffer 21 ml
Ethanol (95% or absolute ) 15 ml
Filter. Useful for 4 to 5 days.
Stain sections 10 to 15 sec. Remove slides and wash in DW. Air dry.
Notes: Works well with methacrylate sections. Epon sections may take longer or
may require heating the staining solution.
Humphrey and Pittman Stain Technology 42:9-14 (1974)
Solutions
A. Methylene blue-azure II
Methylene blue 0.130 gms
Azure II (Azure A can be used) 0.020 gms
Glycerol 10 ml
Methanol 10 ml
PO#031#4 buffer, 0.15 M, pH 6.9 30 ml
DW 50 ml
B. Basic Fuchsin
Stock solution
Basic fuchsin 0.100 gm
50% EtOH 10 ml
Staining solution
Stock solution 3 ml
DW 57 ml
Staining procedure
1. Immerse slides in solution A at 65o C. Time required varies with
thickness and type of embedding media. Ten minutes works for 7 to 10 um
sections of Epon or Epon/Araldite.
2. Rinse in two changes of DW.
3. Stain in solution B at room temperature for 20 min. Again time may vary
with thickness and embedding media.
4. Rinse thoroughly in DW.
5. Dry stained sections on a 40o C. hot plate.
6. Sections can be mounted in Permount. If wrinkling becomes a problem try
mounting in the appropriate resin. Use just enough resin to cover sections. A
thick layer of resin may prevent focusing at high power if your objective has a
short working distance.
Notes:
1. Sorensen's phosphate buffer is used in the original method.
0.15 M Na2HPO4 100 ml
0.15 M KH2PO4 90 ml
pH = 6.9
2. Authors suggest that stain is stable for up to 4 months. The shelf life
in our experience is considerably longer.
3. It is difficult to overstain sections.
Date: Wed, 09 Jun 1999 12:57:18 -0700 (PDT)
From: Katie B <bresee98@yahoo.com>
Subject: Histo in Plastic
To: Histonet Server <histonet@pathology.swmed.edu>
Looking for a recipe for Methylene Blue-Basic Fuchsin stain to do in
GMA embedded tissues. My boss tells me it is the "old" way to do
something like an H+E in plastic.
Thanks!
===
Catherine "Katie" Bresee Bennett
Laboratory for Experimental Pathology
Department of Veterinary Pathology
Michigan State University
e-mail: bresee98@yahoo.com
_________________________________________________________
Do You Yahoo!?
Get your free @yahoo.com address at http://mail.yahoo.com
<< Previous Message | Next Message >>