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Picrosirius red

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Picrosirius red is a stain used in histological procedures to visualize collagen fibers. It binds to collagen and produces a red-orange color under polarized light microscopy, allowing for the identification and assessment of collagen structures in tissue samples.

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Lab products found in correlation

Masson s trichrome, by Merck Group (17 mentions) Ix70 microscope, by Olympus (10 mentions) Alcian blue, by Merck Group (9 mentions) Oil red o, by Merck Group (8 mentions) Masson s trichrome, by Diagnostic BioSystems (6 mentions) Sirius red, by Merck Group (6 mentions) Fast green, by Merck Group (6 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (6 mentions) Hematoxylin and eosin, by Merck Group (5 mentions) Direct red 80, by Merck Group (5 mentions) Toluidine blue, by Merck Group (4 mentions) Safranin o, by Merck Group (4 mentions) Metamorph v7, by Molecular Devices (4 mentions) Picrosirius red, by Olympus (3 mentions) Weigert s haematoxylin, by Merck Group (3 mentions) Metamorph software, by Molecular Devices (3 mentions) Vectamount, by Vector Laboratories (3 mentions) Eclipse 80i brightfield microscope, by Nikon (3 mentions) Alcian blue 8gx, by Merck Group (3 mentions) Paraformaldehyde (pfa), by Avantor (2 mentions)

166 protocols using picrosirius red

1

Picrosirius Red Staining of Femoral Bone

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Femoral bone tissues were decalcified in decalcifying solution (Sigma-Aldrich Chemicals) and dehydrated in a graded series of ethanol solutions for 18 h. For the histological staining of picrosirius red, femoral bone tissues were then embedded in paraffin and cut into 7 μm sections in thickness. picrosirius red staining was commercially used for detecting bone collagen. The sections were placed on glass slides, deparaffinated, and hydrated with xylene and graded alcohol. These sections were incubated with picrosirius red (Sigma-Aldrich Chemicals) solution overnight at RT. After each slide was mounted in VectaMount Mounting Medium (Vector Laboratories, Burlingame, CA), images were taken using an Axiomager optical microscope system (Zeiss, Oberkochen, Germany).
Lee E.J., Kim J.L., Gong J.H., Park S.H, & Kang Y.H. (2015). Inhibition of Osteoclast Activation by Phloretin through Disturbing αvβ3 Integrin-c-Src Pathway. BioMed Research International, 2015, 680145.
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2

Intervertebral Disc Degeneration Histology

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Co5/6 (injured) and Co6/7 (intact control) discs and adjacent vertebrae were isolated en bloc immediately after euthanasia. The disc with its surrounding vertebral bodies was fixed with 4% paraformaldehyde for 24 hours. The bone-disc-bone segments were decalcified with a solution consisting of 12.5% EDTA for approximately 1 week, with shaking, until the bony portion was completely decalcified. The tissues were then dehydrated and embedded in paraffin and sectioned to 5μM thickness. For Picro Sirius Red staining, sections were incubated with 0.1% Picro Sirius Red (Sigma) for 45 minutes.
For immunostaining, a rabbit anti-human cadherin was used at 1:200 dilution (Cat#ab76011, Abcam, Cambridge, MA). Note that the antigen used to generate this antibody was a peptide within aa 150–250 (extracellular domain), and the 8C11 monoclonal antibody used here binds to the extracellular domain of N-cadherin between “extracellular cadherin” (EC)3 and EC4 domains.22 (link) The sections were incubated in EDTA buffer at pH 8.0 for 20 minutes to expose the antigen. Biotinylated linking secondary antibody (Vector Laboratories, Burlingame, CA) at 1:200 dilution was used, and color was developed with DAB (DAKO, Santa Clara, CA).
Wei Y., Tian Z., Tower R.J., Gullbrand S., Yao L., Shetye S.S., Mauck R.L., Qin L, & Zhang Y. (2021). The Inner Annulus Fibrosus Encroaches On the Nucleus Pulposus in the Injured Mouse Tail Intervertebral Disc. American journal of physical medicine & rehabilitation, 100(5), 450-457.
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3

Tissue Processing and Staining Techniques

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One half of the sagittally-divided LV or left kidney were fixed in 10% neutral buffered formalin for 24 h at room temperature. The fixed tissues were cut into 5–7 systematic uniform random tissue slabs with a razor blade fractionator and embedded in paraffin blocks with the cut surface down. For Picro Sirius Red staining, a set of 3 µm sections were cut on a microtome from each block and collected on microscope slides. For Periodic acid-Schiff (PAS) staining, another set of sections were cut in pairs with a dissector distance of 30 µm. Picro Sirius Red staining: After deparaffinization, sections were incubated in Wiegert’s iron hematoxylin (Sigma-Aldrich) and then stained in Picro Sirius Red (Sigma-Aldrich) before they were cover slipped with Pertex. PAS staining: sections were deparaffinized and oxidized with 0.5% periodic acid solution followed by incubation with Schiff's reagent. Sections were counterstained in Mayer's hematoxylin and cover slipped with Pertex. Podocin/type IV collagen double fluorescent immunohistochemical staining of kidney sections was performed as previously described22 (link).
Thisted L., Østergaard M.V., Pedersen A.A., Pedersen P.J., Lindsay R.T., Murray A.J., Fink L.N., Pedersen T.X., Secher T., Johansen T.T., Thrane S.T., Skarsfeldt T., Jelsing J., Thomsen M.B, & Zois N.E. (2020). Rat pancreatectomy combined with isoprenaline or uninephrectomy as models of diabetic cardiomyopathy or nephropathy. Scientific Reports, 10, 16130.
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4

Vertebral Disc Decalcification and Picrosirius Red Staining

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The IVDs and portions of the adjacent bony vertebral bodies were isolated immediately after euthanasia. The disc with its surrounding vertebral bodies was fixed with 4% paraformaldehyde for 24 hours. The bone‐disc‐bone segments were decalcified with a solution consisting of 12.5% EDTA for ~1 week, with shaking, until the bony portion was completely decalcified.26 The tissues were then dehydrated and embedded in paraffin and sectioned to 5 μM thickness. For Picrosirius red staining, sections were stained with 0.1% Picrosirius red (Sigma) for 45 minutes. All samples were examined under a light microscope (Nikon) and photographed.
Tian Z., Shofer F.S., Yao L., Sun H., Zhang H., Qin L., Chen Y.H, & Zhang Y. (2020). TNFAIP8 family gene expressions in the mouse tail intervertebral disc injury model. JOR Spine, 3(2), e1093.
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5

Histological and Fluorescence Analysis of Decellularized ECM

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Paraformaldehyde 4% (Merck, Darmstadt, Germany) was applied to prepare specimens for histological and fluorescence studies. After fixation, specimens were dehydrated through a graded series of ethanol, embedded in paraffin (Lab-O-Wax, Milan, Italy), cross-sectioned at a thickness of 5 µm with a microtome (Leits, Vienna, Austria), deparaffinized, rehydrated and stained with hematoxylin-eosin (H&E) (Merck, Darmstadt, Germany) to determine the construct cellularity. Safranin-O (Merck, Darmstadt, Germany) (25 (link)) and toluidine blue (Merck, Darmstadt, Germany) stainings were performed to demonstrate the GAGs content in decellularized ECM. Picrosirius-red (Merck, Darmstadt, Germany) staining was also employed to examine the effects of decellularization on the collagen-rich ECM. Using polarizing microscope (Olympus, IX70, Japan) for observing the sections stained with Picrosirius-red is a specific method to identify the collagenous structures (26 (link)). DNA contents were stained with 4, 6 diamidino-2-phenylindole (DAPI; Sigma-Aldrich, Taufkirchen, Germany), which is a fluorescent dye that binds to nucleic acids with high affinity for DNA.
Tavassoli A., Matin M.M., Niaki M.A., Mahdavi-Shahri N, & Shahabipour F. (2015). Mesenchymal stem cells can survive on the extracellular matrix-derived decellularized bovine articular cartilage scaffold. Iranian Journal of Basic Medical Sciences, 18(12), 1221-1227.
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6

Histological Analysis of Vertebral Morphology

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Sections used for histology were fixed in 4 % paraformaldehyde (PFA) (VWR, Lutterworth, UK) for 20 min, stained with 0.025 % alcian blue (Merck) in 3 % acetic acid (Merck) (for cartilage) for 30 min followed by 1 % picrosirius red (Merck) (for collagen) for 30 min (Fig. 1c) (Rolfe et al., 2017 (link)). Sections from the cervical, thoracic, and lumbar regions were imaged in transmitted illumination using a light microscope (Yenway EX30, Glasgow, UK), with light intensity being adjusted to observe the vertebral bodies. Vertebral segmentation, disc morphology, and vertebral shape were qualitatively compared between control and mdg spines, using light microscopy images.
Levillain A., Ahmed S., Kaimaki D.M., Schuler S., Barros S., Labonte D., Iatridis J.C, & Nowlan N.C. (2021). PRENATAL MUSCLE FORCES ARE NECESSARY FOR VERTEBRAL SEGMENTATION AND DISC STRUCTURE, BUT NOT FOR NOTOCHORD INVOLUTION IN MICE. European cells & materials, 41, 558-575.
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7

Cardiac Biomarkers and Oxidative Stress

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Isoprenaline hydrochloride was purchased from Sigma-Aldrich (3050 Spruce St. 63103 St. Louis, USA). Standards and all other reagents for MDA, NO, APOP assays, and Picrosirius red staining reagents were purchased from Merck (Darmstadt, Germany) and Sigma-Aldrich (3050 Spruce St. 63103 St. Louis, USA). SOD standard and other assay components were purchased from SR Group (Delhi, India). Assay kits for creatinine kinase-muscle brain (CK-MB), uric acid and creatinine were purchased from DCI diagnostics (Budapest, Hungary). GeneJET RNA Purification Kit, RevertAid First Strand cDNA Synthesis Kit (Catalog number: K1621) and SYBR™ Green PCR Master Mix were purchased from Thermo Fisher Scientific Inc. (Waltham, Massachusetts, United State of America). l-carnitine was received from General Pharmaceuticals Pvt. Ltd, Dhaka, Bangladesh as a gift sample.
Chisty T.T., Sarif S., Jahan I., Ismail I.N., Chowdhury F.I., Siddiqua S., Yasmin T., Islam M.N., Khan F., Subhan N, & Alam M.A. (2024). Protective effects of l-carnitine on isoprenaline -induced heart and kidney dysfunctions: Modulation of inflammation and oxidative stress-related gene expression in rats. Heliyon, 10(3), e25057.
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8

Histological Analysis of Vertebral Morphology

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Sections used for histology were fixed in 4 % paraformaldehyde (PFA) (VWR, Lutterworth, UK) for 20 min, stained with 0.025 % alcian blue (Merck) in 3 % acetic acid (Merck) (for cartilage) for 30 min followed by 1 % picrosirius red (Merck) (for collagen) for 30 min (Fig. 1c) (Rolfe et al., 2017 (link)). Sections from the cervical, thoracic, and lumbar regions were imaged in transmitted illumination using a light microscope (Yenway EX30, Glasgow, UK), with light intensity being adjusted to observe the vertebral bodies. Vertebral segmentation, disc morphology, and vertebral shape were qualitatively compared between control and mdg spines, using light microscopy images.
Levillain A., Ahmed S., Kaimaki D.M., Schuler S., Barros S., Labonte D., Iatridis J.C, & Nowlan N.C. (2021). PRENATAL MUSCLE FORCES ARE NECESSARY FOR VERTEBRAL SEGMENTATION AND DISC STRUCTURE, BUT NOT FOR NOTOCHORD INVOLUTION IN MICE. European cells & materials, 41, 558-575.
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9

Renal Histopathology and Lipid Evaluation

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To evaluate morphological changes, left renal tissues were fixed in 10% formalin and embedded in paraffin. Coronal sections were collected from left kidney for histopathological examination via hematoxylin and eosin staining (Solarbio, China). To assess fibrosis, renal tissue sections were stained with picrosirius red (Sigma-Aldrich) [24 (link)]. Lipid accumulation in renal tissue sections was evaluated by Oil Red O staining [25 ]. Images were observed using light microscopy (magnification 400 × , Nikon, Tokyo, Japan) and analyzed by ImageJ software. Images of the cortex and medulla (3–4 images per region) from four mice in each group were analyzed.
Han F., Kan C., Wu D., Kuang Z., Song H., Luo Y., Zhang L., Hou N, & Sun X. (2022). Irisin protects against obesity-related chronic kidney disease by regulating perirenal adipose tissue function in obese mice. Lipids in Health and Disease, 21, 115.
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10

Histological Analysis of Atherosclerotic Plaques

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Paraffin sections of human atherosclerotic plaques were de-waxed, hydrated and stained with Weigerts haematoxylin (Sigma, St Louis, MO) for 8 minutes prior to staining with Picro Sirius Red (Sigma; 0.1% in saturated picric acid solution) for 1 hour. Sections were washed in acidified water, dehydrated and mounted in eukitt. Pictures were obtained in a microscope with circularly polarized light.
Abbas A., Aukrust P., Russell D., Krohg-Sørensen K., Almås T., Bundgaard D., Bjerkeli V., Sagen E.L., Michelsen A.E., Dahl T.B., Holm S., Ueland T., Skjelland M, & Halvorsen B. (2014). Matrix Metalloproteinase 7 Is Associated with Symptomatic Lesions and Adverse Events in Patients with Carotid Atherosclerosis. PLoS ONE, 9(1), e84935.
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