RE: Masson alternative

From:James Hall <>


I think you have hit the nail on the head.  The senior technician from whom
I obtained the method worked at St.Andrews University, as did I at the
time, and Professor Lendrum and his chief technician, Bill Slidders, worked
at Dundee University, approximately 8 miles away.  Prof Lendrum had an
arrangement with the industrial dye Companies, including ICI, that any new
dye they produced they would send him samples to try out on histology
sections.  We had close links with Prof Lendrum and he would pass on
samples and methods for us to try out at St.Andrews.  I must admit I had
totally forgotten about this link, sign of old age, thanks for jogging my
memory.  It would be interesting to find out if this method was ever
published as an alternative to the Masson.


At 08:35 26/09/00 -0400, Hewlett Bryan wrote:
>     Jim,   This method appears to be a slight variation of the Picro-
>Mallory V method of Lendrum and Slidders (1962). See Studies on the
>character and staining of fibrin. Lendrum, A.C., Fraser, D.S., Slidders, W.
>and Henderson, R. J.Clin. Pathol., 15,401-413.  Bryan   ---------- 
>From:  James Hall[] 
>Sent:  September 26, 2000 6:29 AM 
>Subject:       Masson alternative   Hi Folks,   The following method is an
>alternative to the standard Masson technique and 
>has the advantage that it can be carried out on formalin fixed tissues 
>without the need to post chrome or fix in a mercury containing fixative 
>  It is a method which was passed on to me in the 60's by my senior at 
>  I have no idea as 
>to the origins of the technique, perhaps John can come up with the answer 
>to this, but it is the method I prefer to the Massons and it worked 
>  Hopefully some of you will give it a try 
>and I would be grateful for any feedback.  
>@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@   Masson trichrome using a yellow
>mordant.   Fixation - routine fixatives containing mercury are recommended
>but formol 
>saline is satisfactory however fresh material is essential.   Solutions:- 
>1. Celestine blue and Mayer's haemalum for nuclear staining.   2. Yellow
>Stock solution 1 - 100 ml saturated solution of picric acid in 70% alcohol 
>  Dissolve the orange G in a small amount of 
>distilled water and add to the picric acid solution. 
>Stock solution 2 - 0.4g of lissamine fast yellow in 40 ml distilled water. 
>Add solution 1 to 2 and store at room temperature.   3. Ponceau 2R (ponceau
>de xylidine) 2g, plus 1g acid fuchsin, plus 200 ml 
>distilled water plus 3 ml acetic acid.   4. 1% phosphomolybdic acid.   5.
>1% acetic acid   6. Connective tissue counterstain - 1% light green or
>methyl blue in 1% 
>acetic acid (1% direct sky blue in 1% acetic acid is another alternative)  
>1. Take 6 micron sections to water via the iodine/ sodium thiosulphate 
>sequence if necessary. 
>2. Stain nuclei in filtered celestine blue for 10 minutes, rinse in tap 
>water then 10 minutes in filtered Mayer's haemalum followed by a few 
>minutes in running tap water. 
>3. Stain in a working solution of the yellow mordant for 3 to 5 minutes 
>     (ie working solution is 30 ml of the combined solutions 1 and 2 in 70 
>ml of 80% alcohol). 
>4. Wash in running tap water until the yellow colour has been extracted 
>from the muscle - approx 1 minute. 
>5. Stain in the ponceau acid fuchsin solution for 5 minutes. 
>6. Rinse off the excess dye in tap water for approx. 20 seconds. 
>7. Place sections in 1% phophomolybdic acid until the red colouration is 
>retained in the muscle only and can be up to 30 minutes depending on the 
>freshness of the tissue and the fixation. 
>8. Place in the connective tissue stain (fresh material which has had 30 
>minutes in the phophomolybdic acid give 2 minutes, other material not so 
>well fixed which has had only 1 to 5 minutes in the phophomolybdic acid 
>  5 minutes for the direct sky blue irrespective of the 
>time in phophomolybdic acid. 
>9. Rinse well in 1% acetic acid followed by 75% alcohol. 
>10. Dehydrate clear and mount.   Results:_ 
>Nuclei - blue/black 
>Muscle - red 
>Elastic tissue - pale pink 
>RBC's - yellow 
>Collagen, reticulin and mucin - green or blue depending on counterstain
>  Jim Hall, 
>MDA Equipment Evaluator, 
>Department of Histopathology, 
>University College London Hospitals, 
>Rockefeller Building, 
>University Street, 
>London, WC1E 6JJ. 
>  020 7679 6042 
>  020 7387 3674 
>Web Site:    

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