Description 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. VitroView™ Picro-Sirius Red Stain Kit Introduction Video Kit Components 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 Storage Condition Room temperature. Protocol 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. Expected results Light Microscopy Collagen ————————————————————– red Muscle Fibers —————————————————– yellow Cytoplasm ———————————————————- yellow Polarized Light Microscopy Type I (Thick fibers) —————————————- yellow‐orange birefringence Type III (Thin fibers) ——————- green birefringence More Pico-Sirius Red Staining Images Picro-Sirius Red Staining, Rat Liver (Fibrosis, 20x) Picro-Sirius Red Staining, Rat Liver (Fibrosis, 2x) Picro-Sirius Red Staining, Rat Liver (Fibrosis, 0.2x) Picro-Sirius Red Staining, Rat Liver (10x) Picro-Sirius Red Staining, Mouse Mammary-gland, 20x Picro-Sirius Red Staining, Xenograft Tumor, 20x Picro-Sirius Red Staining, Mouse Liver, 4x Picro-Sirius Red Staining, Mouse Liver, 40x Picro-Sirius Red Staining, Mouse Liver, 40x 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. Peer-reviewed publications and preprints that have cited the use of VitroView™ Picro-Sirius Red Stain Kit (VB-3017) Kin J (2023)-TLR7 activation by miR-21 promotes renal fibrosis by activating the pro-inflammatory signaling pathway in tubule epithelial cellsCell Communication and Signaling (2023) 21:215 — This study used VitroView™ Picro-Sirius Red Stain Kit (VB-3017) to visualize collagen I and III in kidney sections to assess the degree of renal fibrosis. Springer Nature Link Kim YY, et al (2022) — Genetic Deletion of Syndecan-4 Alters Body Composition, Metabolic Responses, and Collagen Deposition in High-Fat Diet-Induced Obesity.Nutrients 11(11):2810. Staining of Type I and III collagen fibers was performed with a Picro-Sirius Red staining kit (Cat. No. VB-3017, VitroVivo Biotech, Rockville, MD, USA) to assess collagen deposition in adipose tissues. MDPI Ha S, et al (2022)-Activation of PAR2 promotes high-fat diet-induced renal injury by inducing oxidative stress and inflammationInternational Journal of Molecular Sciences / ScienceDirect 1868 (10), 166474— Sirius Red staining was performed using a commercially available kit from VitroVivo (VB-3017) to determine renal fibrosis and damage. ScienceDirect Marini V, et al (2022)-Long-term culture of patient-derived cardiac organoids recapitulated Duchenne muscular dystrophy cardiomyopathy and disease progressionPMC (National Institutes of Health) 10.3389/fcell.2022.878311 — This article describes using the VitroView™ Picro-Sirius Red Stain Kit (VB-3017) to evaluate collagen deposition in cardio-related organoid tissue (used according to manufacturer’s instructions). PMC Giarratana et al (2020)-MICAL2 is essential for myogenic lineage commitment (PMC) Cell Death and Disease (2020) 11:654 — Picro-Sirius Red staining was carried out using Vitro View™ Picro-Sirius Red Stain Kit (VB-3017) in mouse tissue sections to dye connective tissue collagen. PMC Lu A, et al (2025)-Muscle-specific ERRγ activation mitigates muscle atrophy after ACL injury FASEB Journal 39 (4): e70409 — This research reports H&E and Picro-Sirius Red stain (VitroView™ VB-3017, VitroVivo Biotech) staining following the manufacturer’s protocol for fibrosis/collagen visualization in muscle tissue. FASEB Journal User Manual and Material Safety Data Sheet (MSDS) (PDF) VB-3017 User manual VB-3017 MSDS
