This is a quick version of Movat’s Pentachrome. The long version is an overnight stain, I hate waiting till the next day to find out if the stain worked. It is sometimes hard to visualize all 5 colors but with a aorta in the heart, it is possible. The stain picture will have to be edited in later.
Control tissue that I use: Skin, Aorta in the heart
Safety equipment: Work under a hood with lab coat, gloves, and glasses.
- Deparaffinize and hydrate to DH2O
- Preheat Alkaline alcohol in 60C oven or water bath
- Mordant in bouin’s at 60C water bath——- 30 minutes (under hood!) (re-use)
- Cool—————————————– 10 minutes in room temp water
- Wash in running water—————– 10 minutes
- 1% Alcian blue————————— 25 minutes (re-use)
- Wash in running water—————– 5 minutes
- Alkaline alcohol at 60C —————- 10 minutes
- Wash in running water—————— 5 minutes
- Weigerts———————————– 15 minutes (dump)
- Differentiate in 2% ferric chloride for- 3 minutes (re-use)
- Rinse in H2O——————————- 10 seconds
- Sodium thiosulfate for——————– 1 minute (re-use)
- Wash in running water——————- 5 minutes
- Crocein scarlet/ Acid fuchsin———- 1.5 minutes (re-use)
- Wash in running water—————— 5 minutes
- 5% Phosphotungstic acid————– 5 minutes (re-use)
- Transfer directly to 1% acetic acid– 5 minutes (re-use)
- Wash in running water—————— 5 minutes
- Dehydrate quickly 95% alc. 1 min/2x 100% alc. 1 min
- Alcoholic saffron————————– 60 minutes (re-use)
- Rinse in 2 changes of 100% alcohol, 1 min each
- Clear in xylene and mount
Nuclei and elastic fibers—Black
Ground substance and mucin–Blue
Fibrinoid, fibrin–Intense Red
An Expert Histology Service
42 thoughts on “Movat’s Modified Pentachrome Protocol”
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Great article! I am relatively new to Histology and I was wondering if this stain can be de-stained and repeated if the slides didn’t stain well? I tried the Masson de-stain protocol and it worked great! thanks!
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