Specific staining by Picro-sirius red is one of the most important stains to study collagen networks in different tissues. VitroView™ Picro-Sirius Red Stain Kit is designed for stain of collagen I and III fibers. The Sirius Red stain may be viewed using standard light microscopy or polarized light resulting in birefringence of the collagen fibers to distinguish between type I and type III.
- VB-3017-1 Weigert’s Hematoxylin Solution A——–125 ml
- VB-3017-2 Weigert’s Hematoxylin Solution B ——-125 ml
- VB-3017-3 Picro-sirius Red Solution———————-250 ml
- VB-3017-4 Acidified Watetr————————————250 ml
- a).For FFPE slides: Deparaffinize in Xylene I for 6 minutes and II for 6 minutes. Rehydrate: ethanol 100% (2 minutes)×2; ethanol 95% (2 minutes)×2; ethanol 70% (2 minutes)×1; b). For Frozen Sections: fix frozen sections in 10% formalin for 30 minutes.
- Rinses in distilled water 2min×3.
- Prepare Weigert’s working hematoxylin: mix Weigert’s Hematoxylin Solution A and B at 1:1 ratio.
- Stain nuclei with Weigert’s haematoxylin for 8 minutes, and then wash the slides for 10 minutes in running tap water.
- Stain in Picro-sirius Red Solution for 1 hour. Note: This gives near-equilibrium staining, which does not increase with longer times. Shorter times should not be used, even if the colors look good.
- Wash in two changes of acidified water.
- Wash slides, 2min × 2 in dH2O.
- Dehydrate with 2 changes of 95% Ethanol and 2 changes of 100% Ethanol (2 minute per change).
- Clear with 3 changes of xylene (5 minute per change) and coverslip with Permount or other suitable organic mounting medium.
- Collagen ————————————————————– red
- Muscle Fibers —————————————————– yellow
- Cytoplasm ———————————————————- yellow
Polarized Light Microscopy
- Type I (Thick fibers) —————————————- yellow‐orange
- birefringence Type III (Thin fibers) ——————- green birefringence
Note: VitroView™ Picro-Sirius Red Stain Kit is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Precautions: Handle with care. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.