Re: Sirius Red. (Solution & method)

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From:"J. A. Kiernan" <> (by way of histonet)
To:histonet <>
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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

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