Picrosirius red

Manufactured by ScyTek Laboratories

Sourced in United States

Picrosirius red is a staining dye used in histological analysis. It selectively stains collagen fibers, allowing for the visualization and assessment of collagen content in tissue samples.

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Lab products found in correlation

Picro sirius red, by Molecular Devices (2 mentions) Bouin s fixative, by Electron Microscopy Sciences (2 mentions) Picric acid, by Merck Group (2 mentions) Spectramax 250, by Molecular Devices (2 mentions) Histovt one, by Nacalai Tesque (1 mentions) Optimal cutting temperature compound, by Sakura Finetek (1 mentions) Bz x analyzer, by Keyence (1 mentions) Bz x700, by Keyence (1 mentions) E 600, by Olympus (1 mentions) Fv1000, by Olympus (1 mentions) Toluidine blue, by ScyTek Laboratories (1 mentions) Masson s trichrome, by ScyTek Laboratories (1 mentions) 24 well plate, by Corning (1 mentions)

7 protocols using picrosirius red

Wound Healing Macroscopic and Histopathological Analysis

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After euthanasia, at 7 or 14 days, a macroscopic analysis of the lesions on the dorsum of the rats was performed. The percentage area of the wound on the postoperative days was calculated by comparing the changes in wound size in relation to the first day of the procedure. The monitoring of cicatricial evolution was used in ImageJ software for measurement (de Avila Santana et al., 2013 (link)). After the macroscopic analysis, wounds were excised with 2 mm margin beyond the wound edge for histopathological evaluation, and a representative sample was used to illustrate the wound healing observed in the present study.
For histopathological analysis, tissue samples were fixed with 10% neutral buffered formalin, dehydrated with alcohol at different times and concentrations, embedded in paraffin and cutted at 5 μmin a microtome for staining with routine hematoxylin-eosin (HE) (Abramov et al., 2007 (link)), Mallory’s trichrome (Cheng et al., 2008 (link)) or picrosirius red (ScyTek^®, Logan, UT, United States) (Gowda et al., 2017 (link)). The sections were examined under a light microscope (E400, Nikon, Tokyo, Japan). To evaluate birefringence pattern of collagen fibers, the sections were examined under 100× magnification using a polarized light microscope.

de Aragão Tavares E., de Medeiros W.M., de Assis Pontes T.P., Barbosa M.M., de Araújo A.A., de Araújo RF J.r., Figueiredo J.G., Leitão R.C., da Silva Martins C., da Silva F.O., de Brito Pontes A.C., de Lima Pontes D, & de Medeiros C.A. (2019). Chitosan Membrane Modified With a New Zinc(II)-Vanillin Complex Improves Skin Wound Healing in Diabetic Rats. Frontiers in Pharmacology, 9, 1511.

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Histological Analysis of Liver Tissue

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Tissues were fixed with 4% paraformaldehyde, embedded into optimal cutting temperature compound (Sakura Finetek), and cryosectioned at a thickness of 5 μm. For immunohistochemistry analysis, antigen retrieval was performed using HistoVT One (Nacalai Tesque) at 70 °C for 20 min. Specimens were blocked with 5% BSA and then incubated with the appropriate primary antibodies (Table S2) dissolved in PBS containing 1% BSA. Specimens were then labeled with an appropriate fluorescent secondary antibody. Oil Red O staining was performed to determine lipid accumulation in liver tissues. Sirius red staining (Picro Sirius Red; ScyTek) was performed to evaluate hepatic fibrosis. Images were acquired using a fluorescent microscope (Nikon) and analyzed using ImageJ software (NIH).

Akasu R., Miyazaki T., Elhussiny M.Z., Sugiura Y., Tomitsuka Y., Haraguchi S., Otsu K., Chowdhury V.S, & Miyazaki A. (2022). Calpain-mediated proteolytic production of free amino acids in vascular endothelial cells augments obesity-induced hepatic steatosis. The Journal of Biological Chemistry, 298(6), 101953.

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Histomorphometric Analysis of Osteoclast Activity

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After μCT analyses, maxillary samples were decalcified in EDT-X (Falma, Tokyo, JP) for 3 weeks and embedded in paraffin after dehydration. Four μm sections were made centered on the second molar site. Sections were stained with TRAP^{61 (link)–63 (link)}, and Picro-Sirius Red (ScyTek, Logan, US) respectively. The bone surface area around the second molar root was defined as the bone surface for normalization. TRAP-positive multinucleated (> 3 nuclei) cells in the bone surface area were counted as osteoclasts, and the crenated or lacunar bone surface area was defined as the eroded surface. All images were obtained by the all-in-one fluorescence microscope BZ-X700 (KEYENCE, Osaka, JP), and the eroded surface and bone surface were measured with a BZ-X analyzer (v.1.4.0.1; KEYENCE).

Liu A., Hayashi M., Ohsugi Y., Katagiri S., Akira S., Iwata T, & Nakashima T. (2024). The IL-33/ST2 axis is protective against acute inflammation during the course of periodontitis. Nature Communications, 15, 2707.

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Quantifying Collagen Deposition in Differentiated Cells

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The total collagen deposition of the differentiated cells was evaluated at the cell density of 3 × 10³ cells/cm². Samples were fixed in Bouin’s solution (Bouin’s Fixative, Electron Microscopy Sciences, USA) for 1 h and collagen fibers stained with 0.1% Picro-Sirius Red (ScyTek Laboratories) saturated in picric acid (Sigma-Aldrich). The collagen matrix deposition was finally visualized under polarized light microscopy^{23 (link)}. After staining, the Picro-Sirius Red from the stained cells were extracted with a 0.2 M NAOH and methanol (1:1) buffer to quantify the stained collagen, and the OD of the extract was measured at 550 nm using an ELISA reader (Spectra MAX 250; Molecular Devices, Sunnyvale, CA).

Yeh S.H., Yu J.H., Chou P.H., Wu S.H., Liao Y.T., Huang Y.C., Chen T.M, & Wang J.P. (2024). Proliferation and Differentiation Potential of Bone Marrow–Derived Mesenchymal Stem Cells From Children With Polydactyly and Adults With Basal Joint Arthritis. Cell Transplantation, 33, 09636897231221878.

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Assessing Collagen Fibrillar Organization

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Fibrillar collagen organization was assessed using both polarized light microscopy and second-harmonic generation (SHG) imaging. For polarized light analysis, samples were stained with picrosirius red (ScyTek) to enhance collagen birefringence and imaged using a polarized light microscope (Olympus E600) equipped with rotating polarizer/analyzer (images acquired at 10° increments; mean pixel intensity over entire μPP pattern area measured for each sample). To detect collagen alignment for a given pattern architecture, differences between mean pattern intensities with polarizer angle were tested (repeated measures ANOVA). To detect differences between pattern architectures, mean pattern intensities at a given polarizer angle were compared (t-test). Bonferroni corrections were made for multiple comparisons. SHG image stacks of samples (2 μm deep, starting just above pattern surface) were acquired using a two-photon microscope (Olympus FV1000, 25X objective, Ex/Em:880/440 nm). Directionality of SHG images was analyzed via FFT as described above, with image AI calculated about the histogram mode or pattern orientation.

Gilchrist C.L., Ruch D.S., Little D, & Guilak F. (2014). Micro-scale and meso-scale architectural cues cooperate and compete to direct aligned tissue formation. Biomaterials, 35(38), 10015-10024.

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Tissue Embedding and Staining Protocol

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Collected samples were fixed in 4% paraformaldehyde at 4 °C overnight, embedded in paraffin, and cut into sections (3 μm thick), including the cross section of embedded silicone tube in the rostral direction. Deparaffinized sections were stained with hematoxylin-eosin, Masson's trichrome, toluidine blue, or picrosirius red (ScyTek Laboratories; Logan, UT, USA).

Oe S., Masum M.A., Ichii O., Nishimura T., Nakamura T., Namba T., Otani Y., Nakayama Y., Elewa Y.H.A, & Kon Y. (2021). Spatiotemporal histological changes observed in mouse subcutaneous tissues during the foreign body reaction to silicone. Journal of biomedical materials research. Part A, 109(7).

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Quantifying Collagen Deposition in Cells

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Collagen deposition of the differentiated cells was evaluated using picrosirius red staining. The cells were seeded in a 24-well plate (Corning Life Science) at a density of 3 × 10 3 cells/cm 2 , fixed in Bouin's solution (Bouin's Fixative; Electron Microscopy Sciences, Hatfield, PA, USA) for 1 hour, and stained with 0.1% picrosirius red (ScyTek Laboratories) saturated in picric acid (Sigma-Aldrich). Collagen deposition was visualized under polarized light microscopy. 24 Quantification of the stained collagen was detected by extracting the picrosirius red from the stained cells with a 0.2 M NaOH and methanol (1:1) buffer, and the OD of the extract was measured at 550 nm using an ELISA reader (Spectra MAX 250; Molecular Devices).

Huang H.K., Hsueh K.K., Liao Y.T., Wu S.H., Chou P.H., Yeh S.H, & Wang J.P. (2023). Multilineage differentiation potential in the infant adipose- and umbilical cord-derived mesenchymal stem cells. Journal of the Chinese Medical Association : JCMA, 86(12).

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