We've used this protocol in the past with some success on 15Ám ground MMA sections. It was adapted from a Nottingham University method. As usual, you may need to tweak the times to suit your sections.
Smith &Nephew Research Centre
York Science Park
* Ponceau/Acid Fuchsin Stock Solution:
1.5 g - Ponceau 2R.
0.5 g Acid Fuchsin.
2 ml Acetic Acid (glacial).
98 ml Distilled Water.
* Azophloxine Stock Solution:
0.5 g Acid Red 1.
0.6 ml Acetic Acid (glacial).
99.4ml Distilled Water.
* Ponceau Working Solution:
12ml Ponceau/Acid Fuchsin Stock Solution.
8ml Azophloxine Stock Solution.
80 ml 0.2% Acetic Acid.
* Orange G Solution:
1 g Orange G.
1.5g Phosphomolybdic Acid.
250 ml Distilled Water.
* Light Green Solution:
1 g Light Green (Yellowish).
1 ml Acetic Acid (glacial).
500 ml Distilled Water.
1) Slides in distilled water for 15 minutes.
2) Stain in Weigert's Haematoxylin for 30 minutes.
3) Wash in running tap water for 2 minutes.
4) Differentiate in 0.5% acid alcohol (30 seconds).
5) Wash in water for 20 minutes.
6) Stain in Ponceau Working Solution for 5 minutes.
7) Rinse in 1% Acetic Acid for 15 seconds.
8) Stain in Orange G Solution for 20 minutes.
9) Rinse in 1% Acetic Acid for 10 seconds.
10) Stain in Light Green Solution for 20 minutes.
11) Rinse in water.
12) Blot dry.
Nuclei = blue/black.
Mineralised bone = green.
Muscle = green.
Osteoid = red.
Collagen = red.
From: email@example.com [mailto:firstname.lastname@example.org] On Behalf Of email@example.com
Sent: 27 February 2008 18:05
Subject: Histonet Digest, Vol 51, Issue 40
Date: Wed, 27 Feb 2008 10:19:52 -0500
Subject: [Histonet] goldners trichrome
Content-Type: text/plain; charset=US-ASCII
I was asked to do a Goldners Trichrome in Methymethacylate embedded bone .
Do you have a good protocol for this or do you know where I could find one.
Thank you for any help you can give me.
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