Accustain trichrome stain kit

Manufactured by Merck Group

Sourced in Germany

The Accustain Trichrome Stain Kit is a laboratory reagent used for the histological staining of tissue samples. The kit provides the necessary dyes and solutions to perform a trichrome staining technique, which is commonly used to differentiate between various tissue components, such as collagen, muscle, and nuclei. The specific details and intended use of the product are not available.

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Lab products found in correlation

Apoptosis in situ detection kit, by Fujifilm (1 mentions) Cm1900, by Leica (1 mentions) Refrigerated microtome, by Leica (1 mentions) Masson trichrome staining, by Merck Group (1 mentions) Mayer hematoxylin, by Bio-Optica (1 mentions) Axioscop2 mot plus microscope, by Zeiss (1 mentions) Aniline blue, by Merck Group (1 mentions) Phosphomolybdic phosphotungstic acid solution, by Merck Group (1 mentions) Weigert s iron hematoxylin set, by Merck Group (1 mentions) Permount, by Thermo Fisher Scientific (1 mentions) Bouin s solution, by Merck Group (1 mentions) K3468, by Agilent Technologies (1 mentions) Dako antibody diluent, by Agilent Technologies (1 mentions) Anti ad2 5 e1a antibody, by Santa Cruz Biotechnology (1 mentions) Bhabp, by AMS Biotechnology (1 mentions) Dpvr110hrp, by Immunologic (1 mentions) Immpact amec red peroxidase hrp substrate, by Vector Laboratories (1 mentions) Vectastain elite abc hrp kit, by Vector Laboratories (1 mentions) Oct compound, by Sakura Finetek (1 mentions)

5 protocols using accustain trichrome stain kit

Histological Assessment of Interstitial Fibrosis

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Cross sections (10 μm) were cut with a cryostat (CM1900, Leica Microsystems, Nussloch, Germany) at -20 ºC. The sections were air-dried and fixed with 4% paraformaldehyde (v/v) in 0.1 M phosphate-buffered saline (pH 7.5)^{43 (link),63 (link),64 (link)}.
Interstitial fibrosis was evaluated by Masson-trichrome staining using the Accustain Trichrome Stain Kit (#HT15-1KT; Sigma-Aldrich) in accordance with the manufacturer’s protocol as described previously^{22 (link)}. Interstitial fibrotic regions were quantified using Image J 1.48v software (National Institute of Health, Bethesda, MD, USA) to determine the percentage of blue-stained area in the Masson-trichrome sections^{22 (link),43 (link),64 (link)}. DNA fragmentation was determined by TUNEL staining using an Apoptosis in situ Detection Kit (#293-71501; Wako, Osaka, Japan). The total number of TUNEL-positive nuclei was counted manually in six sections of three groups (Control (n = 4), BO (n = 5), and Pro + BP (n = 4)) over a microscopic field of 20 x, averaged, and expressed as the ratio of TUNEL-positive nuclei (%)^{20 (link),22 (link),43 (link),64 (link)}.

Suita K., Yagisawa Y., Ohnuki Y., Umeki D., Nariyama M., Ito A., Hayakawa Y., Matsuo I., Mototani Y., Saeki Y, & Okumura S. (2020). Effects of occlusal disharmony on susceptibility to atrial fibrillation in mice. Scientific Reports, 10, 13765.

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Histological Analysis of Liver Tissue

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The left lobe of the liver was fixed in 10% neutral formalin solution (Sigma–Aldrich) overnight at 4 °C, cryopreserved in a 30% (w/v) sucrose solution for 24 h at 4 °C, and stored at −80 °C. Liver sections 7-μm thick were cut with a refrigerated microtome (Leica), collected on slides, and stored at −80 °C until staining. Hematoxylin–eosin (H&E) staining was performed on the frozen slides with Mayer hematoxylin (Bio-Optica) for 1 min and, after washing with water, with 1% eosin aqueous solution (Bio-Optica) for 4 min. Oil Red O staining was performed as previously described [1 (link)]. The Accustain Trichrome stain kit was used for Masson trichrome staining (Sigma–Aldrich). After staining, the slides were cleared in xylenes and cover slipped with xylene-based mounting medium (Eukitt, Bio-Optica). The liver sections were evaluated in a blinded fashion under a light microscope. Semi-quantitative grading of the hepatocellular vacuolar degeneration and portal inflammation (i.e., the infiltration of mononuclear inflammatory cells) was also performed in a blinded fashion. Images of the stained sections were captured using Microscope Axioscop2 mot plus (Zeiss).

Meda C., Barone M., Mitro N., Lolli F., Pedretti S., Caruso D., Maggi A, & Della Torre S. (2019). Hepatic ERα accounts for sex differences in the ability to cope with an excess of dietary lipids. Molecular Metabolism, 32, 97-108.

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Trichrome Staining of Tissue Sections

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Sections from WT and miR-205^−/− transplanted outgrowths were subjected to a series of steps of deparaffinization and rehydration then mordant in Bouin’s Solution (Millipore Sigma, St. Louis, Missouri, www.emdmillipore.com/US/en/home.html) at room temperature overnight to intensify the final coloration of the tissue. Weigert’s Iron Hematoxylin Set (Sigma) was used to stain nuclei. Sections were then put in Biebrich Scarlet-Acid Fuchsin from Accustain Trichrome Stain Kit (Sigma) for 5 minutes to stain cytoplasm and muscle red. Phosphomolyb-dic/Phosphotungstic Acid Solution (5 minutes) and Aniline Blue (5 minutes) from Accustain Trichrome Stain Kit (sigma) were used to develop collagen blue staining. Upon detection, sections were rinsed in 1% acetic acid solution for 5 minutes, dehydrated, and mounted using Permount (Fisher Scientific).

LU Y., CAO J., NAPOLI M., XIA Z., ZHAO N., CREIGHTON C.J., LI W., CHEN X., FLORES E.R., MCMANUS M.T, & ROSEN J.M. (2018). miR-205 Regulates Basal Cell Identity and Stem Cell Regenerative Potential During Mammary Reconstitution. Stem cells (Dayton, Ohio), 36(12), 1875-1889.

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Immunohistochemical Analysis of E1A, HA, and CD3

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Paraffin-embedded blocks were cut into 4-μm thick sections. E1A and HA staining were performed as previously published,¹⁴ (link)^,²¹ (link) using the anti-Ad2/5 E1A antibody (1/200; Santa Cruz Biotechnology, SC-25) and B-HABP (5 μg/mL; Amsbio, AMS.HKD-BC41). For human CD3 detection, FLEX Polyclonal Rabbit Anti-Human CD3 (IR503, Agilent DAKO) was diluted 1:10 with DAKO antibody diluent (Dako – Agilent, S0809) for 120 min at room temperature. The secondary antibody used was a BrightVision poly-horseradish peroxidase (HRP)-anti-rabbit immunoglobulin G (IgG) that was biotin-free, ready to use (Immunologic, DPVR-110HRP) incubated 45 min. Antigen-antibody complexes were reveled with 3-3′-diaminobenzidine (K3468, Dako). H&E staining was performed according to standard procedures. Masson trichromic staining was performed using the Accustain Trichrome Stain Kit (Sigma Aldrich) according to the manufacturer’s indications. The percentage of stained areas in IHC images were quantified after color deconvolution by ImageJ FIJI⁴⁴ (link) software.

Farrera-Sal M., Moreno R., Mato-Berciano A., Maliandi M.V., Bazan-Peregrino M, & Alemany R. (2021). Hyaluronidase expression within tumors increases virotherapy efficacy and T cell accumulation. Molecular Therapy Oncolytics, 22, 27-35.

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Cardiac Tissue Immunohistochemistry and Trichrome Staining

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For immunohistochemistry, hearts were harvested and embedded in O.C.T compound (Sakura Finetek). Embedded tissues were snap-frozen in dry ice. Sections of 5 µm thickness were then stained using antibodies directed against CD11b (clone M1/70) or CD31 (clone MEC 13.3) (all BioRad). Staining was followed with a biotinylated secondary antibody. We used the VECTASTAIN Elite ABC HRP kit and ImmPACT AMEC Red Peroxidase (HRP) substrate (Vector Laboratories, Inc.) for color development. For Masson’s trichrome staining, we used Weigert’s iron hematoxylin solution and Accustain Trichrome Stain Kit (both Sigma-Aldrich) according to the manufacturer’s protocol.

Seung H., Wrobel J., Wadle C., Bühler T., Chiang D., Rettkowski J., Cabezas-Wallscheid N., Hechler B., Stachon P., Maier A., Weber C., Wolf D., Duerschmied D., Idzko M., Bode C., von zur Mühlen C., Hilgendorf I, & Heidt T. (2022). P2Y12-dependent activation of hematopoietic stem and progenitor cells promotes emergency hematopoiesis after myocardial infarction. Basic Research in Cardiology, 117(1), 16.

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