Movat Pentachrome stain
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Movat Pentachrome Stain Kit (Modified Russell-Movat) is intended for use in histological demonstration of collagen, elastin, muscle, mucin and fibrin in tissue sections. This procedure is particularly useful when studying the heart, blood vessels and various vascular diseases.
1) Deparaffinize sections if necessary and hydrate to distilled water.
2) Stain tissue section with working Elastic Stain Solution for 20 minutes.
3) Rinse in running tap water until no excess stain remains on slide.
4) Dip slide in Ferric Chloride (2%) Differentiating Solution 15-20 times and rinse in tap water.
5) Check slides microscopically for proper differentiation. Repeat step 4 if required.
6) Rinse in 2 changes of distilled water.
7) Place slide in Sodium Thiosulfate Solution (5%) and incubate for 1 minute.
8) Rinse in tap water for 2 minutes followed by 2 changes in distilled water.
9) Place slide in Acetic Acid Solution (3%) and incubate for 2 minutes to equilibrate tissue prior to staining with Alcian Blue Solution, pH 2.5.
10) Without rinsing, place slide in Alcian Blue Solution, pH 2.5 and incubate for 25 minutes.
11) Rinse in tap water for 2 minutes followed by 2 changes in distilled water.
12) Place slide in Biebrich Scarlet – Acid Fuchsin Solution and incubate for 2 minutes.
13) Rinse slide in 2 changes of distilled water.
14) Place slide in Acetic Acid Solution (1%) for 5-10 seconds with agitation.
15) Rinse quickly in distilled water.
16) Differentiate slide in 2 changes of Phosphotungstic Acid Solution (5%) for 3-7 minutes each.
17) Check slides microscopically for proper differentiation. Collagen should be clear but elastic fibers should still be stained. Repeat step 16 if required.
18) Rinse slide briefly in distilled water. 1.20 Incubate slide in Acetic Acid Solution (1%) for 1 minute. This step is important for removing Phosphotungstic acid bound to the tissue.
19) Shake off excess Acetic Acid Solution (1%) and without rinsing apply Metanil Yellow Solution and incubate for 15 minutes.
20) Rinse slide in 3 changes of absolute alcohol.
21) Clear, and mount in synthetic resin.
Elastic Fibers: Black to Blue/Black
Reticular Fibers: Yellow
Mucin: Bright Blue
Fibrin: Bright Red