>Date: Sat, 28 Aug 1999 21:04:48 GMT
>From: AndreaH@imclone.com
>Subject: Masson's Trichrome
>To: "HistoNet Server" <histonet@pathology.swmed.edu>
>Mime-Version: 1.0
>
>
>I would like to successfully stain skin with the Masson's Trichrome
>technique.  I was hoping that I could get some helpful hints from the
>Histonet world about this stain.  Any useful info would be appreciated but
>particularly I would like to know where I should buy the individual
>reagents, how people do the stain regularly nowdays, etc.  Thanks for your
>help in advance.
>
>Andrea
>

Andrea,
	Pretty straightforward stuff. The result for skin will be slightly
different but it gives a very pleasant  picture. You'll get the recipes for
the stains in any Histo-technique text. Any problems let me know. Me, I
like the Celestine Blue /Haemalum (Mayer's haematoxylin) option. Saves
differentiating and I prefer the detail of the nuclear staining. I always
dilute the green. Rather than a jumping out at you green it gives a more
delicate shade, just like the gentle caress of a beautiful woman.
Ian.

MASSON'S TRICHROME.

SOLUTIONS.
1.)  WEIGERT'S HAEMATOXYLIN.
2.)  ACID ALCOHOL.
3.)  CELESTINE BLUE.
4.)  HAEMALUM.
5.)  PONCEAU ACID FUCHSIN.
6.)  5% PHOSPHOMOLYBDIC ACID.
7.)  MASSON'S LIGHT GREEN.

METHOD.
  1.) Bring sections to water.
   Either.
  2.) Weigert's Haematoxylin. = 10  minutes.
  3.) Rinse in water.
  4.) Differentiate in acid alcohol until nuclei only are stained.
  5.) Wash well in water.
   Or.
  2.) Celestine blue.	= 5 minutes.
  3.) Rinse in water.
  4.) Heamalum.	= 5 minutes.
  5.) Blue in running tap water.

  6.) Ponceau acid fuchsin. = 5 minutes.
  7.) Rinse briefly in water.
  8.) Differentiate in phosphomolybdic acid until the connective tissue is
unstained or very pale pink.
DO NOT RINSE IN WATER. Phosphomolybdic acid acts as a mordant for the green.
  9.) Light Green.	= 30 seconds. (Control microscopically.)
10.) Rinse in water.
11.) D.C.M.

RESULT.
	Nuclei.	                   =  Mauve or purple/blue.
	Muscle and Nerve.	=  Red.
	Connective tissue.	=  Green.

NOTES.
	 Stain carefully with light green to avoid overstaining. If
overstained the green can be extracted using dilute acetic acid - can prove
troublesome.
	The best method is dilute the light green 10 fold.

Dr. Ian Montgomery,
West Medical Building,
University of Glasgow,
Glasgow,
G12 8QQ,
Scotland.
Tel: 0141 339 8855 Extn. 6602.
Fax: 0141 330 4100.
e-mail: ian.montgomery@bio.gla.ac.uk