[Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers

[Guillermo Palao]

Thanks, you advice going straight from sirius red to 70% ethanol or maybe to 100% as suggested in other reply, but how do I get rid of the excess red stuff on the slide?
   
  Best regards,
   
  Guillermo

Tony Henwood <AnthonyH <@t> chw.edu.au> escribió:
  Drop step 3 and 4 (picric acid is soluble in water)

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Guillermo Palao
Sent: Wednesday, 5 April 2006 4:59 AM
To: Histonet
Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers


Hello all:

I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes.

My protocol is as follows:

1. Deparafinize and hydrate samples
2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid
3. Wash 2 min 0.01 N HCl
4. Rinse in water
5. Dehydrate and cover slides

I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated.

Guillermo


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