At 4 weeks, the mandible was collected to analyze the multi-tissue regeneration effect. After fixing it with 4% paraformaldehyde for 24 h, the tissue was scanned using a micro-CT scanner (μCT50; SCANCO Medical AG, Switzerland). Materialize Mimics Research 19 (Materialize, Belgium) software was used to reconstruct the three-dimensional images. The mandible was further decalcified in a 10% EDTA solution (pH 7.4) for four weeks and H&E staining was performed. For Goldner's trichrome staining, the slices were first stained with hematoxylin for 20 min, followed by staining with fuchsin acid, orange G, fuchsia acid, and light green stain for 5–10, 1–5, 3–5, and 3–10 min, respectively (Servicebio, China). For picrosirius red staining, the paraffin sections were impregnated with picrosirius red (Servicebio, China) for 10 min and differentiation was induced with ethanol for 3 s. IHC staining of ALP (1:100, Abcam, UK), OPN (1:200, Abcam, UK), and BSP (1:200, Abcam, UK) was performed as previously described. The sections were scanned using an Aperio AT2 system. The picrosirius red-stained sections were observed and photographed using a polarized light microscope.
Picrosirius red
Manufactured by Wuhan Servicebio Technology
Sourced in China
Picrosirius red is a histological stain used for the detection and visualization of collagen fibers in tissue samples. It binds specifically to collagen and produces a red-orange color under polarized light microscopy, allowing for the identification and analysis of collagen-rich structures within the sample.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ci, by Nikon (2 mentions)
Masson trichrome, by Wuhan Servicebio Technology (1 mentions)
Alcian blue, by Wuhan Servicebio Technology (1 mentions)
Periodic acid schiff, by Wuhan Servicebio Technology (1 mentions)
Fsx100 microscope, by Olympus (1 mentions)
Masson s trichrome, by Wuhan Servicebio Technology (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
μct50, by Scanco (1 mentions)
4 protocols using picrosirius red
1
Multi-tissue Regeneration Analysis of Mandible
2
Histological Evaluation of Tissue Samples
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Native and decellularized samples were fixed in 4% paraformaldehyde, dehydrated in graded alcohol series, embedded in paraffin, and sectioned into 5-μm sections. The glass slides were stained with hematoxylin and eosin (H&E, Servicebio, Wuhan, China), Masson trichrome, periodic acid–Schiff, Alcian blue, and picrosirius red (PSR) according to the manufacturer’s instructions. All reagents were purchased from Servicebio. The sections were imaged on a light microscope (CX43, Olympus, Tokyo, Japan). PSR-stained slides were assessed by phase-contrast microscopy (Eclipse Ci, Nikon, Tokyo, Japan) and analyzed by the CT-FIRE program.
3
Quantification of Collagen Fibers in OS
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Picrosirius red staining was used for histological visualization of collagen fibers. Picrosirius red (Servicebio, Wuhan, China)) was used to quantify the percentage of collagen fibers in OS specimens and metastatic tumors in the lung. All the samples were formalin-fixed, paraffin-embedded, cut into sections, and stained with Picrosirius red. The sections were captured by Olympus FSX100 microscope (Olympus, Japan).
4
Histological Evaluation of Tissue Samples
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Tissue samples were fixed in 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, United States) in phosphate-buffered saline (PBS) for 24 h and embedded in paraffin, after which 4-μm-thick sections were stained with hematoxylin and eosin (H&E; Servicebio, Wuhan, China), Picrosirius Red (Servicebio), and Masson’s trichrome (Servicebio) for conventional morphological evaluation after deparaffinization and rehydration. For the measurement of epidermal and dermal thickness, five fields from each section were randomly selected for statistical analysis. Collagen fibers were observed using polarized light microscopy (Eclipse Ci; Nikon, Tokyo, Japan), with collagen I fibers stained red and newly synthesized fibers stained yellow to orange (16 (link)), whereas collagen III fibers appeared green. Collagen volume fraction (CVF) was used to assess the relative content of collagen in the dermis stained with Masson’s trichrome.
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