The fixed muscles were dehydrated and embedded in paraffin. Six-micrometer-thick sections were cut. The sections were stored at 4 °C until staining. After deparaffinization and hydration of the sections, the sections underwent staining. For hematoxylin and eosin staining, the sections were stained with hematoxylin (Muto Pure Chemicals) for 3 min followed by 2 min of staining with eosin (Wako). For modified Gomori’s trichrome staining, the sections were stained with hematoxylin (Muto Pure Chemicals) for 3 min followed by 10 min of staining with modified Gomori’s trichrome staining solution (Chromotrope 2 R, 9.6 mg·mL−1; Fast green FCF, 4.8 mg·mL−1; phosphotungstic acid hydrate 9.6 mg·mL−1; and acetic acid, 16 μL [pH 3.4]). The histological images were captured, and muscle fiber width and tendon width (the minor axes) were measured using measurement software (BZ–X analyzer, Keyence).50 (link)
Hematoxylin
Manufactured by Muto Pure Chemicals
Sourced in Japan, United States
Hematoxylin is a natural dye derived from the heartwood of the Haematoxylum campechianum tree. It is a widely used staining agent in histology and cytology laboratories. Hematoxylin stains cell nuclei and other cellular components, enabling visualization and identification of various tissue structures under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Biotinylated anti mouse igg antibody, by Vector Laboratories (5 mentions)
Eclipse e200, by Nikon (5 mentions)
3 3 diaminobenzidine (dab), by Thermo Fisher Scientific (4 mentions)
Skimmed milk, by BD (3 mentions)
Oil red o, by Merck Group (3 mentions)
Hydrogen peroxide, by Fujifilm (3 mentions)
Vectastain abc kit, by Vector Laboratories (2 mentions)
Oct compound, by Sakura Finetek (2 mentions)
Diaminobenzidine, by Agilent Technologies (2 mentions)
Real envision kit, by Agilent Technologies (2 mentions)
Glycerol gelatin, by Merck Group (2 mentions)
Diaminobenzidine tetrahydrochloride dab staining reagent, by Agilent Technologies (2 mentions)
Phosphate buffered saline pbs, by Fujifilm (2 mentions)
Alcian blue solution, by Muto Pure Chemicals (1 mentions)
Malinol, by Muto Pure Chemicals (1 mentions)
Diaminobenzidine tetrahydrochloride, by Nacalai Tesque (1 mentions)
Avidin biotin blocking kit, by Nichirei Biosciences (1 mentions)
Acetone, by Fujifilm (1 mentions)
Avidin biotin blocking kit, by Vector Laboratories (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
46 protocols using hematoxylin
1
Histological Evaluation of Muscle and Tendon
2
Alcian Blue Staining of Lung Tissue
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Frozen lung sections were rinsed with 3% acetic acid and then stained with Alcian blue solution (pH 2.5, Muto Pure Chemicals, Tokyo, Japan) for 20 min at room temperature (approximately 25 °C). After washing with water for 1 min, tissues were incubated with 3% acetic acid for 3 min, washed with water for 1 min, and nuclei were stained with hematoxylin (Muto Pure Chemicals) for 2 min. After washing with water for 10 min, the sections were dehydrated using a graded ethanol series (70, 80 90, 95, and 100%), defatted with xylene twice, and mounted in Malinol (Muto Pure Chemicals).
3
Tissue Distribution of SBSN Peptides
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An antibody cross-reactivity tissue array, MNO341 (US Biomax, Rockville, MD, USA), which contained 33 tissue types mostly from surgical resection, was used to examine the tissue distribution of each peptide. The tissue preparations were deparaffinized, rehydrated, and autoclaved in pH 6.0 citric acid buffer for antigen retrieval. The preparations were blocked using an endogenous Avidin Biotin Blocking Kit (Nichirei Bioscience, Tokyo, Japan) and then incubated overnight at 4 ℃ with either anti-SBSN_HUMAN[225–237] IgG, anti-SBSN_HUMAN[243–259] IgG, anti-SBSN_HUMAN[279–295] IgG or control non-immune IgG purified from preimmune serum (1:1000 dilution). After washing in PBS, the samples were incubated with a biotinylated anti-rabbit IgG antibody (1:3000, Abcam) for 60 min at room temperature. Antibody binding was visualized by the avidin–biotin-complex peroxidase method using the Vectastain ABC kit (Vector Laboratories, Burlingame, CA, USA) with diaminobenzidine tetrahydrochloride (Nacalai Tesque). The tissue preparations were stained with hematoxylin (Muto Pure Chemicals CO, Tokyo, Japan)60 (link).
4
Immunohistochemical Analysis of hMUC1 in Breast Cancer
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Example 15
Paraffin-embedded human breast cancer tissue sections were purchased from ISU ABXIS (Seoul, Korea). The tissue sections were treated with xylene for 30 minutes to remove paraffin, rehydrated with ethanol, and incubated with 3% hydrogen peroxide solution for 10 minutes. Antigen retrieval was performed in a citric acid solution (pH 6.0). The sections were blocked with normal horse serum for 30 minutes and incubated with anti-hMUC1 antibody (1 μg/slide) at room temperature for 2 hours. The sections were washed and incubated with a biotinylated anti-mouse IgG antibody (Vector Laboratories, Burlingame, Calif.) for 1 hour. Then, the tissue sections were washed and incubated with HRP-streptavidin for 30 minutes. Immunoreactivity was detected using 3,3-diaminobenzidine (DAB, Thermo Fisher Scientific) and the tissue sections were counterstained with hematoxylin (Muto Pure Chemicals, Tokyo, Japan). All images were examined using a microscope (Eclipse E-200, Nikon, Tokyo, Japan).
5
Immunohistochemical Analysis of PDAC
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A human PDAC tissue section (OriGene Technologies, Rockville, MD, USA) and a normal human pancreatic tissue section (BioChain, Newark, CA, USA) were purchased commercially. Resected pancreatic cancer tissues were embedded in an optimal-cutting-temperature (OCT) compound (Sakura Finetek Japan, Tokyo, Japan) and were frozen at −80 °C. Frozen section samples were fixed using chilled acetone (Wako Pure Chemical, Osaka, Japan). In the staining of human PDAC and normal pancreas sections, after blocking with 5% skimmed milk (Becton Dickinson, Franklin Lakes, NJ, USA) at room temperature for 30 min, the section was incubated with HRP-conjugated 102-10 IgG or isotype control IgG, anti-CD20 antibody for overnight at 4 °C. The staining of mouse PDAC and normal pancreas sections were conducted with VECTASTAIN Elite ABC KIT (Vector Laboratories, Burlingame, CA, USA) according to the manufacturer’s instructions. Endogenous biotin of pancreas was blocked by using Avidin/Biotin Blocking Kit (Vector Laboratories). All reactions were visualized using a diaminobenzidine staining reagent (Dako, Glostrup, Denmark), and the samples were counterstained with hematoxylin (Muto pure chemicals, Tokyo, Japan).
6
Histological Analysis of Cochlea
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Cochlea sections on slides were stained with hematoxylin (Muto Pure Chemicals Co. Ltd.) and eosin (Wako Pure Chemicals Industries Ltd., Osaka, Japan) (HE staining) to study their structures. Cochlear specimens were examined using a light microscope system (BZ-8100, Keyence, Osaka, Japan) and saved as digital images.
7
Hematoxylin and Eosin Staining of Tumor-Associated Macrophages
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For H&E staining, M0, TAM(Ca), TAM(CAF) and TAM(TAM) were seeded in the Nunc Lab-Tek II Chamber Slide system (cat. no. 154526PK; Thermo Fisher Scientific, Inc.). The subsequent steps were all performed at room temperature. The slides were gently washed twice with PBS and fixed with 90% ethanol for 15 min. Then, the slides were stained with hematoxylin (Muto Pure Chemicals Co., Ltd.) for 10 min, washed under running water for 5 min and stained with eosin Y (FUJIFILM Wako Pure Chemical Corporation) for 5 min. After washing under running water for 5 min to remove the excess dye, the slides were sealed with a coverslip and neutral resin. Differences between each group were compared under a light microscope (magnification, ×200).
8
Comprehensive Tissue Sampling for Histopathology
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Tissue sampling was conducted on day 0 (before administration of ABC), 1, 3, 5, or 7, respectively. The brain, liver, heart, lung, kidney, thymus, spleen, cervical and mesenteric lymph nodes, and skin (auricle and back) were fixed in 10% neutral buffered formalin (Nacarai Tesque; Kyoto, Japan). These tissues were sectioned, embedded in paraffin (Sakura Finetek; Tokyo, Japan), and stained with hematoxylin (Muto Pure Chemicals; Tokyo, Japan) and eosin (Sakura Finetek) using a routine procedure. All histopathological evaluations were conducted by a board-certified veterinary pathologist.
9
Immunohistochemical Analysis of PGRMC1 in Mouse Ovaries
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Ovaries of 4-week-old female mice were fixed with SUPER FIX (KURABO, Osaka, Japan) and embedded in paraffin. The embedded ovary was sliced into 6-μm thick serial sections. Endogenous peroxidase activity was blocked by treatment
with 0.6% hydrogen peroxide/methanol at room temperature for 40 min. Sections were treated with anti-PGRMC1 antibody (2 μg/ml; Abcam, Cambridge, UK) or rabbit IgG (2 μg/ml; Vector Laboratories, Burlingame, CA, USA) and subjected
to immunoperoxidase staining using the VECTASTAIN ABC Kit (Vector Laboratories) according to the standard protocol. All sections were counterstained with hematoxylin (MUTO PURE CHEMICALS, Tokyo, Japan).
with 0.6% hydrogen peroxide/methanol at room temperature for 40 min. Sections were treated with anti-PGRMC1 antibody (2 μg/ml; Abcam, Cambridge, UK) or rabbit IgG (2 μg/ml; Vector Laboratories, Burlingame, CA, USA) and subjected
to immunoperoxidase staining using the VECTASTAIN ABC Kit (Vector Laboratories) according to the standard protocol. All sections were counterstained with hematoxylin (MUTO PURE CHEMICALS, Tokyo, Japan).
10
Immunohistochemical Analysis of hMUC1 in Breast Cancer
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Example 15
Paraffin-embedded human breast cancer tissue sections were purchased from ISU ABXIS (Seoul, Korea). The tissue sections were treated with xylene for 30 minutes to remove paraffin, rehydrated with ethanol, and incubated with 3% hydrogen peroxide solution for 10 minutes. Antigen retrieval was performed in a citric acid solution (pH 6.0). The sections were blocked with normal horse serum for 30 minutes and incubated with anti-hMUC1 antibody (1 μg/slide) at room temperature for 2 hours. The sections were washed and incubated with a biotinylated anti-mouse IgG antibody (Vector Laboratories, Burlingame, Calif.) for 1 hour. Then, the tissue sections were washed and incubated with HRP-streptavidin for 30 minutes. Immunoreactivity was detected using 3,3-diaminobenzidine (DAB, Thermo Fisher Scientific) and the tissue sections were counterstained with hematoxylin (Muto Pure Chemicals, Tokyo, Japan). All images were examined using a microscope (Eclipse E-200, Nikon, Tokyo, Japan).
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