EMS26385
Components
Alcian Blue, 1%
Alkaline Alcohol
Orcein, 0.2%
Haematoxylin Alcoholic, 5%
Ferric Chloride, 10%
Lugol’s Iodine
Woodstain Scarlet-Acid Fuchsin Working Solution
Acetic Acid, 0.5%
Phosphotungstic Acid, 5%
Alcoholic Saffron
Fixation
10% Buffered Neutral Formalin or Bouin's Solution
Note: If formalin fixative is used, decerated tissue sections must be mordanted in Bouin's Fluid for 1 hour in a 50°C oven. Wash well in running water to remove picric acid deposits.
Sections
4-6µm
Staining Procedure
- Decerate section and hydrate to distilled water.
- Stain in Alcian Blue, 1% for 20 minutes.
- Dip five times in distilled water.
- Place in Alkaline Alcohol in a 56°C oven for 10 minutes.
- Wash in running tap water for 2 minutes.
- Stain in Orcein-Verhoeff working solution for 2 hours.
To prepare Orcein-Verhoeff working solution:
Immediately before use, in the following order, combine the solutions below:
Orcein | 0.2% 25.0ml |
Alcoholic Haematoxylin | 5% 8.0ml |
Ferric Chloride | 10% 5.0ml |
Lugol's Iodine | 5.0ml |
- Wash in running tap water for 3 minutes.
- Stain in Woodstain Scarlet- Acid Fuchsin working solution for 2 1/2 minutes.
- Place in Acetic Acid, 0.5% for 30 seconds.
- Differentiate in Phosphotungstic Acid, 5% for 5-10 minutes, well differentiated sections demonstrate colourless collagen and blue-green mucopolysaccharides.
- Rinse in Acetic Acid, 0.5% for 30 seconds.
- Three changes of 100% ethyl alcohol, 1 minute each,
- Stain in Alcoholic Saffron for 8 minutes.
- Dehydrate in 100% ethyl alcohol, two changes.
- Clear in xylene, three changes.
- Coverslip.
Stain Results
Nuclei | Black |
Cytoplasm | Red |
Elastic Fibres | Purple to Black |
Collagen and Bone | Yellow |
Mucopolysaccharides | Blue-Green |
Muscle | Red |
References
Movat, H.Z.,: Demonstration of all Connective Tissue Elements in a Single Section.
Arch Path., 60:289-295, 1955.
Roman N., et al: Orcein-Hematoxylin in Iodized Ferric Chloride as a Stain for Elastic Tissue.
Stain Techn., 42:199-202, 1967.
Silverman Jefferey., A Modified Movat Pentachrome Stain. Histologic , p. 33. 1972
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