Masson Trichrome Variants

In the comparison chart below, all solids are in grams and all liquids are in millilitres with the exception of sat., which means a saturated solution. In each case, dissolve the reagents in distilled water, and make the final volume up to 100 mL.

For more details of each Masson variant, please select the header for each column.

Solution Reagent Gen V-A1 V-A2 V-A3 V-A4 V-B V-C Lillie
Plasma stain Acid fuchsin
Ponceau 2R
Biebrich scarlet
Acetic acid
0.5
0.5
 
1.0
0.5
 
 
0.5
0.35
0.65
 
1.0
 
1.0
 
1.0
1.0
1.0
 
1.0
0.1
 
 
 
1.0
 
 
1.0
 
 
0.5
1.0
Polyacid Molybdophosphoric acid
Tungstophosphoric acid
1.0
 
1.0
 
1.0
 
1.0
 
1.0
 
1.0
 
1.0
 
2.5
2.5
Fibre stain Light green SF yellowish
Aniline blue
Fast green FCF
Metanil yellow
Phosphomolybdic acid
Acetic acid
2.0
 
 
 
 
2.0
 
sat.
 
 
 
2.5
 
sat.
 
 
 
2.5
 
sat.
 
 
 
2.5
 
sat.
 
 
 
2.5
 
0.5
 
 
0.5
 
 
 
 
sat.
 
 
 
 
2.5
 
 
2.5

 

Staining
In the chart below the numbers refer to the time in minutes for which each solution should be applied. As with most staining techniques, these times are recommendations and should be altered if necessary to optimise results. Please consult the full method for more details.

Most trichrome stains benefit from fixation with picric acid or mercuric chloride containing fixatives. If simple formalin fixation has been used, sections may be beneficially treated with a saturated aqueous picric acid solution or Bouin's picro-formalin-acetic mixture for an hour at 56°C.

Method

  1. Bring sections to water via xylene and ethanol.
  2. Treat with picric acid or Bouin's fluid if wished.
  3. Stain nuclei with an acid resistant nuclear stain (Weigert, celestine blue hemalum).
  4. Wash with water. Rinse well with distilled water.
  5. Continue according to the chart below.
  Gen V-A (all) V-B V-C Lillie
6 Plasma stain 10 5 5 (50°C) 5 2
7 Polyacid 5 5 5-10 5 1
8 Fibre stain 10 2-5 20-60 5 2
9 Distilled water Rinse   Rinse Rinse  
10 1% acetic acid water   Differentiate   5 1

 

  1. Dehydrate with ethanol, clear with xylene and mount using a resinous medium

Expected results


 

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