VitroView Movat Pentachrome Stain Kit (#VB-3023)
Description
The VitroView™ Movat Pentachrome Stain Kit (Modified Russel-Movat ) is intended for use in histological visualization of collagen, muscle tissue, reticular fibers, mucins and fibrin in tissue sections. This kit is a very useful tool to study the heart, blood vessels and various vascular diseases.
Kit Components
| SKU# | Reagent Name | Volume (ml) |
| VB-3023-1 | Alcoholic Hematoxylin (5%) | 250 |
| VB-3023-2 | Ferric Chloride Solution (10%) | 125 |
| VB-3023-3 | Iodine Solution | 125 |
| VB-3023-4 | Ferric Chloride Solution (2%) | 125 |
| VB-3023-5 | Sodium Thiosulfate Solution (5%) | 125 |
| VB-3023-6 | Acetic Acid Solution (3%) | 125 |
| VB-3023-7 | Alcian Blue Solution, pH 2.5 | 125 |
| VB-3023-8 | Biebrich Scarlet–Acid Fuchsin Solution | 125 |
| VB-3023-9 | Acetic Acid Solution (1%) | 250 |
| VB-3023-10 | Phosphotungstic Acid Solution (5%) | 250 |
| VB-3023-11 | Metanil Yellow Solution | 125 |
Storage
Room temperature.
Protocol
Things to do before starting the procedures:
Prepare a working Hematoxylin Solution by mixing 30 ml of Hematoxylin (5%) Solution, 15 ml of Ferric Chloride Solution (10%) and 15 mL of Iodine Solution. Note: Mixed solution may be used for 24 hours.
Procedures for FFPE tissue sections
- Deparaffinize in Xylene I for 6 minutes and II for 6 minutes.
- Rehydrate: Ethanol 100% (2 minutes); Ethanol 100% (2 minutes); Ethanol 95% (2 minutes); Ethanol 95% (2 minutes); Ethanol 70% (2 minutes).
- Rinse in distilled water (5 minutes).
- Stain tissue section with working Hematoxylin Solution for 20 minutes.
- Rinse in running tap water until no excess stain remains on slide.
- Dip slide in Ferric Chloride Solution (2%) 15-20 times and rinse in tap water
- Check slides microscopically for proper differentiation. Repeat step 3-6 if required.
- Rinse in 2 changes of distilled water.
- Place slide in Sodium Thiosulfate Solution (5%) and incubate for 1 minute.
- Rinse in tap water for 2 minutes followed by 2 changes in distilled water.
- 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.
- Without rinsing, place slide in Alcian Blue Solution, pH 2.5 and incubate for 25 minutes. Rinse in tap water for 2 minutes followed by 2 changes in distilled water.
- Place slide in Biebrich Scarlet–Acid Fuchsin Solution and incubate for 2 minutes.
- Rinse slide in 2 changes of distilled water.
- Place slide in Acetic Acid Solution (1%) for 5-10 seconds with agitation.
- Rinse quickly in distilled water.
- Differentiate slide in 2 changes of Phosphotungstic Acid Solution (5%) for 3-7 minutes each.
- Check slides microscopically for proper differentiation.
- Collagen should be clear but elastic fibers should still be stained. Repeat step 17 if required.
- Rinse slide briefly in distilled water.
- Dip slide several times (3-5) in Acetic Acid Solution (1%).
- Shake off excess Acetic Acid Solution (1%) and without rinsing apply Metanil Yellow Solution and incubate for 10 minutes.
- Rinse slide in 2 changes of absolute alcohol (1 min per change)
- Clear with 2 changes of xylene (5 minutes per change) and coverslip with Permount or other suitable organic mounting medium.
Expected results
| Nuclei and Elastic Fibers | Black to blue black |
| Ground Substance and Mucin | Bright blue |
| Muscle | Red |
| Collagen | Yellow to red |
| Reticular Fibers | Yellow |
| Fibrinioid and Fibrin | Intense Red |
Control Tissue
Lung, skin, colon, heart or any vascular tissue.