Masson trichrome

Manufactured by Wuhan Servicebio Technology

Sourced in China

The Masson trichrome stain is a histological technique used to identify and differentiate collagen fibers in biological samples. It stains collagen fibers blue, nuclei black, and cytoplasm and muscle fibers red. The stain is commonly used in the analysis of connective tissue and fibrotic conditions.

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12 protocols using masson trichrome

Macrophage Phenotypes in Dental Samples

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Specimens were fixed with 4% paraformaldehyde and decalcified (EDTA, Servicebio, China) for 5–6 weeks at room temperature, then dehydrated. Histological sections were cut buccolingually for HE (Servicebio, China), Masson trichrome (Servicebio, China) and immunohistochemistry staining. Antibodies against inducible nitric oxide synthase (iNOS, Servicebio, China), CD163 (Servicebio, China), CD206 (Abcam, USA), TNF-α (Servicebio, China), and IL-10 (Servicebio, China) were used in the present experiment.
Sections were scanned using Pannoramic MIDI (3DHISTECH Ltd., Budapest, Hungary) and were browsed with CaseViewer software (3DHISTECH Ltd., Budapest, Hungary). For sections of every sample, pictures of certain area were captured by CaseViewer and were used for analysis. The number of iNOS+, CD163+, and CD206+ cells were counted manually in the fixed size region whose location was selected randomly in the captured field (ImageJ). For IL-10 and TNF-α staining, the positively stained area in the field was identified and quantified using preset constant value of color threshold (ImageJ).

Liu J., Chen B., Bao J., Zhang Y., Lei L, & Yan F. (2019). Macrophage polarization in periodontal ligament stem cells enhanced periodontal regeneration. Stem Cell Research & Therapy, 10, 320.

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Histological Analysis of Cardiac Tissue

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The heart tissues were embedded in paraffin wax after fixing in 4% paraformaldehyde for 24 h. Paraffin-embedded heart tissues were cut into 5 µm sections. Hematoxylin and eosin (HE; Sigma, United States) and Masson trichrome (Servicebio, China) were used to observe cardiac histopathological changes according to the manufacturer’s protocol. Briefly, cardiac sections were dewaxed to water and stained. Sections were then dehydrated, made transparent, and sealed. The morphological changes were observed and photographed using light microscopy (Leica, Germany).

Yang H.X., Sun J.H., Yao T.T., Li Y., Xu G.R., Zhang C., Liu X.C., Zhou W.W., Song Q.H., Zhang Y, & Li A.Y. (2021). Bellidifolin Ameliorates Isoprenaline-Induced Myocardial Fibrosis by Regulating TGF-β1/Smads and p38 Signaling and Preventing NR4A1 Cytoplasmic Localization. Frontiers in Pharmacology, 12, 644886.

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Skin Tissue Regeneration Assay

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Skin tissue samples excised 12 days after wounding were fixed in 4% paraformaldehyde for 24 h, embedded in paraffin, then sliced into 5 μm-thick sections. Epidermal regeneration was assessed by detecting collagen in sections stained with hematoxylin and eosin (HE) and Masson trichrome (both from Servicebio Wuhan, China) as described by the manufacturer. Histologically stained tissues were digitally recorded using a DS-Ri2 inverted microscope (Nikon Corp., Tokyo, Japan), and the thickness of the granulation tissue was measured using Image-Pro Plus 6 software (Media Cybernetics, Rockville, MD, USA).

Liu C., Wang Y., Wang P., Gong Y., Yi B., Ruan J, & Wang X. (2023). In situ electrospun aloe-nanofiber membrane for chronic wound healing. Smart Materials in Medicine, 4, 514-521.

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Endometrial Fibrosis Quantification

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Paraffin sections were made for tissue specimens fixed in 4% neutral buffered formalin. These were stained with H&E (cat. no. G1005; Servicebio) and Masson trichrome (cat. no. G1006; Servicebio) according to the protocol provided by the manufacturer and then scanned using a three-dimensional panoramic scanner (3D HISTECH Pannoramic 250; made in Hungary). The number of endometrial glands present in each H&E section of endometrial tissue was counted under a ×40 microscope. Three fields of each Masson section were randomly selected under high magnification (×100), and the image analysis system, Image-Pro Plus 6.0, was used to calculate the area of endometrial stromal fibrosis. The fibrosis ratio was calculated as the area of the endometrial stromal fibrosis in each field divided by the total area of the endometrial interstitium and glands.

Liu N.N., Zhao X., Tan J.C., Liu S., Li B.W., Xu W.X., Peng L., Gu P., Li W., Shapiro R., Zheng X., Zhao W., Jiang Y.G., Chen D., Xu D, & Wang H. (2022). Mycobiome Dysbiosis in Women with Intrauterine Adhesions. Microbiology Spectrum, 10(4), e01324-22.

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Histological Analysis of Scar and Normal Tissue

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Freshly harvested scar and normal tissue were fixed in 4% paraformaldehyde at 4°C for over 24 h, dehydrated and embedded in paraffin following standard protocols by Servicebio, Inc. (Wuhan Servicebio Technology Co., Ltd., Wuhan, Hubei, China). The samples were stored and sectioned at room temperature; 4 µm sections were incubated in a 65°C oven for 2 h and de-paraffinized by washing with xylene and a descending alcohol gradient (100, 100, 85 and 75% for immunohistochemistry staining; 100, 95, 90, 80 and 70% for Masson trichrome staining) prior to staining.
Immunohistochemistry and Masson trichrome staining were performed using mouse-anti-human monoclonal transforming growth factor (TGF)-β1 antibodies (1:200, cat. no. GB11179, Servicebio, Inc., Wuhan Servicebio Technology Co., Ltd.) and Masson trichrome (cat. no. PT003; Shanghai Bogoo Biotechnology. Co., Ltd., Shanghai, China) following the manufacturer's protocols. Secondary antibody staining involved the DAKO REAL EnVision Detection System, Peroxidase/DAB, Rabbit/Mouse (cat. no. K5007; Agilent Technologies, Inc., Santa Clara, CA, USA) and a Masson staining kit (cat. no. G1006; Servicebio, Inc., Wuhan Servicebio Technology Co., Ltd.) according to the manufacturer's protocols.

Zhang K., Chen J., Zhang D., Wang L., Zhao W., Lin D.Y., Chen R., Xie H., Hu X., Fang X, & Fu Q. (2018). microRNA expression profiles of scar and normal tissue from patients with posterior urethral stricture caused by pelvic fracture urethral distraction defects. International Journal of Molecular Medicine, 41(5), 2733-2743.

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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.

Wu T., Gao Y.Y., Tang X.N., Zhang J.J, & Wang S.X. (2022). Construction of Artificial Ovaries with Decellularized Porcine Scaffold and Its Elicited Immune Response after Xenotransplantation in Mice. Journal of Functional Biomaterials, 13(4), 165.

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Decalcification and Histology of Skull

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After micro-CT imaging, the entire skulls were decalcified using 10% ethylenediaminetetraacetic acid (EDTA) solution. Serial sections with 5 mm thickness were prepared along the coronal plane and stained using hematoxylin and eosin (HE) (Servicebio, Wuhan, China) and Masson trichrome (Servicebio). Three central sections were randomly selected along the coronal plane for further histomorphometric measurements.

Meng D., Song J., Yi Y., Li J., Zhang T., Shu Y, & Wu X. (2023). Controlled released naringin-loaded liposome/sucrose acetate isobutyrate hybrid depot for osteogenesis in vitro and in vivo. Frontiers in Bioengineering and Biotechnology, 10, 1097178.

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Quantitative Analysis of Pancreatic Fibrosis

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Pancreas tissue was quickly removed after mice were deeply anesthetized with pentobarbital. After weighing the pancreatic tissue, they were rapidly rinsed thoroughly with phosphate buffered saline for sectioning, pancreas pieces were fixed in 4% paraformaldehyde. The degree of pancreatic fibrosis was evaluated by hematoxylin and eosin (H&E), Masson trichrome and Sirius red staining (performed by Servicebio technology). We quantified the pancreatic fibrosis by calculating the positive area in Sirius red staining.

Li M., Yuan Y., Han X., Liu X., Zhang W, & Hao J. (2022). Antioxidant Mitoquinone Alleviates Chronic Pancreatitis via Anti-Fibrotic and Antioxidant Effects. Journal of Inflammation Research, 15, 4409-4420.

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Histopathological Analysis of Stomach Specimens

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Stomach specimens were dissected along the greater curvature. After washing with phosphate buffer saline, specimens were fixed in paraformaldehyde, embedded in paraffin, and sliced into 4 μm sections. Each sample was stained with haematoxylin–eosin (H&E) (G1005, Servicebio, Wuhan, China) and Masson trichrome (G1006, Servicebio, Wuhan, China) according to the manufacturer s instructions. Moreover, terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL) (In Situ Cell Death Detection Kit, Roche, Basel, Switzerland) and the expression of E-cadherin (1:200) (GB12082, Servicebio, Wuhan, China), β-Catenin (1:200) (G12015, Servicebio, Wuhan, China), CD117 (1:2000) (GB11073-2, Servicebio, Wuhan, China), PGP9.5 (1:100) (GB11159-1, Servicebio, Wuhan, China), and proliferating cell nuclear antigen (PCNA) (1:100) (GB11010, Servicebio, Wuhan, China) were detected by immunofluorescence following the manufacturer’s instructions. Histopathological analysis was performed by two experienced pathologists unaware of the group allocation.

Zhang Y., Han X., Li Z., Zhang Y., Liang L., Ma X., Liu H., Gao Y., Li Q., Chen X., Lv Y, & Ren F. (2021). Physiological and histopathological effects of electroporation pulse on stomach of rats. BMC Gastroenterology, 21, 351.

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Endometrial Fibrosis Quantification

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