[Histonet] question about sirius red stain

[Tony Henwood]

Several thoughts:

Why stain for so long? Try shortening the time to even a few minutes.
The yellow is probably leaching out in the acid water rinse. Try rinsing
in ethanol before clearing and mounting.

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 Xilong
Li
Sent: Saturday, 26 August 2006 3:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] question about sirius red stain


Hi, All,

I stain collagen of frozen muscle section using regular sirius red
staining protocol(0.5g sirius red F3B in 500ml saturated picric acid,
section were stained 1h in solution, then wash in 0.5% acidified water
shortly). The background of images was all red under ordinary
microscopy, but not yellow as expected, collagen can be seen barely with
dark red. I extended washing time in acidified water, but no improvement
was gotten. I just wonder any one had similar experience and how to
improve it?

Thanks in advance.



Dr. Xilong Li
Hypertension Division, Internal Medicine
University of Texas Southwestern Medical Center 
5323 Harry Hiness Blvd-J4.142
Dallas, TX 75390

Tel: 214-648-9966(L)
Fax: 214-648-7902


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