Re: Sirius Red. (Solution & method)
<< Previous Message | Next Message >>
From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
On Thu, 4 Mar 1999, Ian Montgomery wrote:
> This has been asked before, I took details of the technique and
> used it very successfully. But, guess who, foolishly, didn't enter it into
> his techniques book. Please, can the method for Sirius Red be posted again.
> The reference would also be nice.
You can stain nuclei first with iron haematoxylin
or another acid-resisting nuclear stain.
For collagen fibres:
0.1% sirius red F3B in saturated aqueous
picric acid. There should be a little
undissolved picric in the bottom of the
bottle. Solution keeps for years.
Stain for 30-60 minutes. (It is much
slower than Van Gieson's.) Rinse in
slightly acidified water (e.g. 0.1%
acetic acid); dehydrate in 3 X 100%
alcohol, then a couple of xylenes and
coverslip. The alcohol extracts some
(eventually all) of the picric acid from
cytoplasm, so don't linger - unless this
is what you want. Avoid tap water and
lower alcohols, which cause some loss
of both dyes, just as with van Gieson.
Colours similar to Van Gieson, but it
shows thinner fibres and basement membranes
better. Bright colours when you look with
crossed polars, but only the fibres (not
basal laminae) are birefringent.
Original method: Puchtler H & Sweat F 1964.
Histochemie 4: 24-34.
Much studied and somewhat improved by:
Junqueira LCU, Bignolas G & Brentani RR 1979.
Histochem J 11: 447-455.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
<< Previous Message | Next Message >>