Blood was collected by left ventricular puncture from mice and was left undisturbed for 1 h at 37 °C and 2 h at 4 °C. Afterward, the samples were centrifuged at 1000 × g for 10 min at 4 °C; the clear upper fractions were aliquoted and stored at -80 °C. Serum ALT and AST levels were measured on an ADVIA 1800 autoanalyzer (Siemens Healthcare Diagnostics, Deerfield, IL, United States). The livers were preserved in 4% paraformaldehyde, paraffin-embedded and sectioned. The liver tissue sections were stained with hematoxylin and eosin (Beyotime Biotechnology, Shanghai, China) for routine histology and 0.1% Sirius Red (Sigma-Aldrich, St. Louis, MO, United States) for collagen evaluation.
Hematoxylin and eosin
Manufactured by Beyotime
Sourced in China
Hematoxylin and eosin (H&E) is a widely used staining method in histology and pathology. Hematoxylin stains cell nuclei blue, while eosin stains cytoplasm and extracellular structures pink or red. This combination of stains allows for the visualization of cellular and tissue structures under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Rm2160 microtome, by Leica (2 mentions)
Bx53 microscope, by Olympus (2 mentions)
C0105, by Beyotime (2 mentions)
Optical microscope, by Olympus (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Sirius red, by Merck Group (1 mentions)
Advia 1800, by Siemens (1 mentions)
Anti occludin, by Abcam (1 mentions)
Alexa fluor 555 secondary antibody, by Thermo Fisher Scientific (1 mentions)
Safranin o fast green, by Solarbio (1 mentions)
Glass slides, by Beyotime (1 mentions)
Rm2235 rotary microtome, by Leica (1 mentions)
Ba 410 microscope, by Motic (1 mentions)
Image pro plus med 6, by Motic (1 mentions)
Masson s trichrome stain, by Wuhan Servicebio Technology (1 mentions)
Chemray 800, by Rayto (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Solarbio (1 mentions)
76 protocols using hematoxylin and eosin
1
Mouse Blood Serum Analysis and Liver Histology
2
Histological Analysis of Distal Ileum
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The distal ileum was collected and sectioned at the thickness of 4 μm. The slides were stained with hematoxylin and eosin (Beyotime Institute of Biotechnology, Haimen, China) at room temperature. Microscopic scoring was scored according to Nadler et al. (11) . The mean score of five fields was recorded as the final score of the rat. Every group included 10 subjects. Two pathologists who were blind to the study design performed the evaluation separately.
Immunofluorescence was used to locate and evaluate occludin expression. Anti-occludin (1:100 dilution; Abcam, Cambridge, MA, USA) was used. The slides were subsequently conjugated with Alexa Fluor 555 secondary antibodies (Invitrogen, Thermo Fisher Scientific, Waltham, MA). The antigen was visualized using a Leica DFC425 fluorescence microscope (Leica Microsystems (Schweiz) AG, Heerbrugg, Switzerland).
Immunofluorescence was used to locate and evaluate occludin expression. Anti-occludin (1:100 dilution; Abcam, Cambridge, MA, USA) was used. The slides were subsequently conjugated with Alexa Fluor 555 secondary antibodies (Invitrogen, Thermo Fisher Scientific, Waltham, MA). The antigen was visualized using a Leica DFC425 fluorescence microscope (Leica Microsystems (Schweiz) AG, Heerbrugg, Switzerland).
3
Knee Osteoarthritis Assessment Protocol
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The complete knees were fixed with 4% PFA (paraformaldehyde) for 48 h, then the joints were decalcified for 2 months in a 10% EDTA (Ethylene Diamine Tetraacetie Acid, #1430, Biofroxx, Guangzhou, China) solution and paraffin-embedded. After staining the coronal sections (3 μm) with HE (hematoxylin and eosin, #C0105S, Beyotime, Shanghai, China) and SO (safranin O/Fast Green, #G1371, Solarbio, Beijing, China), the slides were examined by independent double-blinded investigators to determine the synovitis and Osteoarthritis Research Society International (OARSI) scores. To determine M1 macrophage polarization, the synovium was stained with immunofluorescence.
4
Histological Assessment of Aortic Wall
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The aortic tissue was fixed in 4% paraformaldehyde for 24 h, and embedded in paraffin. Cross-sections of the aortic tissue were made with a Leica RM2160 microtome at six-micron thickness. Sections were then stained with hematoxylin and eosin (Beyotime, Beijing, China) according to the protocol provided by manufacturer. Pictures of the sections were taken with OlympusBX59, and then ImageJ (NIH, United States) was used to assess the thickness of the vascular wall.
5
Hepatic Fibrosis Assessment in Mice
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In a TAA-induced hepatic fibrosis model, the mice were sacrificed under mild ether anesthesia after Fasudil treatment. Then mouse liver specimens were fixed overnight in 4% paraformaldehyde/phosphate-buffered saline (PBS) and embedded in paraffin. Then liver sections were examined and photographed under a light microscope after hematoxylin and eosin (Beyotime, Shanghai, China) staining. The criteria used for scoring fibrosis and inflammation were as follows: Score 0, normal (no visible fibrosis and inflammation); score 1, fibrosis and inflammation present (5%-30%); score 2, mild fibrosis and inflammation (31%-50%); and score 3, severe fibrosis and inflammation (51%-75%).
6
Histological Analysis of Murine Liver
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Liver tissues from the mice were removed and fixed in 10% neutral buffered formalin. The tissues were then rinsed with water, dehydrated with ethanol, subsequently embedded in paraffin, and sectioned into slices (4-µm-thick) using a Leica RM2235 rotary microtome (Leica Microsystems, Germany). The sections were mounted onto glass slides (Beyotime Institute of Biotechnology, China), dewaxed in xylene and ethanol, and stained with hematoxylin and eosin (Beyotime Institute of Biotechnology, China) for histological evaluation using a Motic BA410 microscope (Motic Incorporation, China). Image Pro-Plus Motic Med 6.0 software (Motic Incorporation) was used for image processing.
7
Histological Analysis of Cartilage
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Briefly, the cartilage surface was cut with a surgical blade, fixed in 4% paraformaldehyde, and paraffin-embedded in the coronal position. The paraffin blocks were sectioned at a thickness of 5 μm, and the resulting sections were deparaffinized in xylene, hydrated with graded ethanol, and stained with hematoxylin and eosin (Beyotime). Photographs were taken using the CX41 pathological microscope.
8
Histological Analysis of Hippocampus
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The hippocampus tissues were collected, and immersed in 3.7% formaldehyde (Sigma-Aldrich, St. Louis, MO, USA), followed via dehydration and paraffin embedding. After that, the tissues were cut to 4-μm sections. Next, the sections were deparaffinized, rehydrated, and stained with hematoxylin and eosin (Beyotime, Shanghai, China) for 10 min. The images of hippocampus sections were observed under a microscope (Olympus, Tokyo, Japan).
9
Histological Analysis of Left Ventricle
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The left ventricle was fixed with 4% paraformaldehyde, dehydrated by gradient alcohol, embedded in paraffin, and cut into 4 μm-thick sections. The sections were stained with hematoxylin and eosin (Beyotime Biotechnology, China) according to the manufacturer's recommendations, followed by observation under an optical microscope (Olympus BX51, Olympus).
10
Comprehensive Cardiac Biomarker Analysis
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An automated biochemical analyzer was used to determine the levels of blood glucose, total cholesterol (TC), and triacylglycerol (TG) in fasting plasma (Chemray 800, Rayto, China). We measured plasma BNP concentrations using an ELISA kit (Raybiotech, Norcross, GA, USA). PKG activity assays were performed using the PKG Kinase Enzyme assay kit from Promega.
Heart morphology were stained with hematoxylin and eosin (Beyotime, Jiangsu, China). Staining cardiomyocytes with FITC-labeled WGA (Servicebio, Wuhan, China) allowed us to determine their cross-sectional area. Masson's trichrome stain (Servicebio, Wuhan, China) was used to examine the myocardium's interstitial fibrosis.
Heart morphology were stained with hematoxylin and eosin (Beyotime, Jiangsu, China). Staining cardiomyocytes with FITC-labeled WGA (Servicebio, Wuhan, China) allowed us to determine their cross-sectional area. Masson's trichrome stain (Servicebio, Wuhan, China) was used to examine the myocardium's interstitial fibrosis.
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