Brain tissues were embedded in paraffin and sectioned. Then sections were processed as follows: Xylene for 20 min, two times; 100% ethanol for 5 min, two times; 75% ethanol for 5 min; and tap water rinsing. Sections were subjected to hematoxylin–eosin (HE) and toluidine blue (Nissl) staining. Hematoxylin–Eosin Staining Kit (G1003; Wuhan Servicebio Technology Co., Ltd., Wuhan, China) was used for HE staining, and Toluidine blue staining solution (G1032; Wuhan Servicebio Technology Co., Ltd.) was used for Nissl staining. Sections were finally sealed with neutral gum and examined under an upright optical microscope (NIKON ECLIPSE E100) for image acquisition.
Hematoxylin eosin staining kit
Manufactured by Wuhan Servicebio Technology
Sourced in China
The Hematoxylin-eosin staining kit is a laboratory product used for the staining of biological samples. It consists of two dyes, hematoxylin and eosin, which are applied sequentially to tissue sections or cells to enhance the visualization of cellular structures under a microscope.
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Lab products found in correlation
Bx61 fluorescence microscope, by Olympus (3 mentions)
Toluidine blue staining solution, by Wuhan Servicebio Technology (2 mentions)
Eclipse e100, by Nikon (1 mentions)
Rosiglitazone, by Selleck Chemicals (1 mentions)
P stat1, by Santa Cruz Biotechnology (1 mentions)
Apc anti mouse cd86, by BioLegend (1 mentions)
Stat6, by Santa Cruz Biotechnology (1 mentions)
T0070907, by Selleck Chemicals (1 mentions)
Recombinant rat il 4, by Thermo Fisher Scientific (1 mentions)
Pparγ, by Bioss Antibodies (1 mentions)
P ppar γ, by Bioss Antibodies (1 mentions)
Socs1, by Affinity Biosciences (1 mentions)
P jak2, by Affinity Biosciences (1 mentions)
P stat6, by Affinity Biosciences (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Optical microscope, by Olympus (1 mentions)
Pas staining kit, by Wuhan Servicebio Technology (1 mentions)
Dab substrate kit, by Thermo Fisher Scientific (1 mentions)
Image pro plus 5, by Media Cybernetics (1 mentions)
13 protocols using hematoxylin eosin staining kit
1
Embedding, Staining, and Imaging Brain Tissue
2
Histological Tissue Processing and Staining
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Brain tissues were embedded in paraffin and sectioned. Then sections were processed as follows: Xylene for 20 min, two times; 100% ethanol for 5 min, two times; 75% ethanol for 5 min; and tap water rinsing. Sections were subjected to hematoxylin–eosin (HE) and toluidine blue (Nissl) staining. Hematoxylin–Eosin Staining Kit (G1003; Wuhan Servicebio Technology Co., Ltd., Wuhan, China) was used for HE staining, and Toluidine blue staining solution (G1032; Wuhan Servicebio Technology Co., Ltd.) was used for Nissl staining.
3
Macrophage Polarization Modulation
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Fluvastatin capsules were purchased from Novartis Pharmaceutical Co., Ltd. (Beijing, China). A Masson staining kit was purchased from Leagene Biotechnology Co., Ltd. (Beijing, China). A hematoxylin-eosin staining kit was purchased from Servicebio Technology Co., Ltd. (Wuhan, China). Cell Counting Kit-8 (CCK-8) was obtained from Fcmacs Biotech Co., Ltd. (Nanjing, China). A BCA protein assay kit was purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Rosiglitazone (RSG, selective PPAR-γ agonist) and T0070907 (selective PPAR-γ antagonist) were obtained from Selleck Chemicals (Houston, USA). LPS was purchased from Sigma Chemical (St. Louis, USA). Recombinant rat IL-4 and INF-γ were purchased from PeproTech Inc. (New Jersey, USA). ELISA kits (IL-1, IL-6, TNF-α, IL4, IL-10, IL-13, and TGF-β1) were obtained from JinYiBai Biological Technology (Nanjing, China). The antibodies (JAK2, STAT6, and p-STAT1) were purchased from Santa Cruz Biotechnology (California, USA). The antibodies (PPAR-γ, STAT1, and p-PPAR-γ) were purchased from Bioss (Beijing, China). The antibodies (SOCS1, SOCS3, p-JAK2, and p-STAT6) were purchased from Affinity Biosciences. APC-anti-mouse CD86 was purchased from BioLegend (San Diego, USA). PE-Cy7-anti-mouse CD206 and Intracellular Fixation & Permeabilization set were purchased from Thermo Fisher Scientific (Massachusetts, USA).
4
Histological Analysis of Muscle Tissue
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After slaughter, the LDM was isolated and fixed in fixative purchased from Servicebio (CAT: G1111-100ML) followed by paraffin sectioning to obtain tissue sections. Then, the LDM paraffin sections were stained using a hematoxylin-eosin staining kit (Servicebio, Wuhan, China). In brief, after deparaffinization with xylene, the sections were stained with hematoxylin solution for 5 min. After that, they were immersed in 1% acid ethanol (1% HCl in 70% ethanol) for 10 s and rinsed with running water. Finally, the sections were stained with eosin for 2 to 3 min, dehydrated with pure alcohol and rendered transparent with xylene. The morphological structure and size of the cells were observed with an Olympus BX61 fluorescence microscope (Japan).
5
Histopathological Evaluation of Colon Tissue
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Proximal colon tissues were dissected from the mice for histological analyses. The tissue sample were fixed in 4% paraformaldehyde for more than 24 h at room temperature, dehydrated and embedded in paraffin. The embedded colon tissues were sliced to the thickness of 4 μm. The sections were stained by Hematoxylin & Eosin staining Kit with the manufacture’s protocol (Servicebio, Wuhan, Hubei, China) and the images was captured by an optical microscope (Olympus, Japan). The severity of inflammation and damage in the colon tissue is evaluated by histopathological scoring in the H&E stain colon sections. The histological scores were measured as follows: (0) For no inflammation and damage in the mucosa of colon; (1) For mild edema and inflammation in the mucosa with 1/3 crypts disappeared; (2) For moderate inflammation in the mucosa with 2/3 crypts disappeared; (3) For moderate inflammation in the mucosa with crypts disappeared completely; and (4) For serious damage and destruction in the mucosa of colon.
6
Histological Analysis of Muscle Tissue
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Longissimus dorsi muscle tissue was extracted from the fixative, paraffin-embedded, and sectioned. Paraffin sections were stained using a hematoxylin-eosin staining kit (Servicebio, Wuhan, China). The slides were examined and photographed using an Olympus BX61 fluorescence microscope (Japan). The cell morphological structures were observed.
7
Histological Analysis of EV-A71 Infection in Murine Eyes
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Eyeballs and lacrimal gland were fixed in 4% formaldehyde and embedded in paraffin. The eyeballs tissue was sliced into 4-μM-thick sections, deparaffinized with xylene, and then hydrated in an ethanol gradient. Then, the sections were stained with hematoxylin-eosin staining kit (Servicebio, Wuhan, China). The eyeballs and eyelids collected from mice were fixed in 4% paraformaldehyde, then embedded with paraffin and sectioned into 5 µm thickness. A PAS staining kit (Servicebio, Wuhan, China) was used to stain the sections. For immunohistochemistry analysis, the sections were incubated with anti-Enterovirus A71 antibodies (Abcam, ab36367) at 4°C overnight. Subsequently, the sections were incubated with secondary antibody (Alexa Fluro 488-labeled goat anti-mouse IgG) for 30 min at room temperature. The bound antibody was performed with a DAB substrate kit (Thermo Scientific). Photos were taken of the sections using a light microscope (Leica Microsystems, Wetzlar, Germany).
8
Histological Analysis of Iliac Vessel
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The iliac vessel was freed from the cuff and preserved in 4% paraformaldehyde for 24 h. The specimens were embedded in paraffin and sectioned (5 µm), and the sections were stained with a hematoxylin-eosin staining kit (Servicebio, China). Image-Pro Plus 5.0 software (Media Cybernetics, Inc., Bethesda, MD, USA) was used to delineate the intima and lumen regions.
9
Histological Analysis of Tissue Samples
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The tissues were taken from the fixative solution for paraffin embedding and microtome sectioning. The paraffin sections were stained by hematoxylin-eosin staining kit (Servicebio, Wuhan, China). Briefly, after deparaffinization and rehydration, sections were stained with hematoxylin solution for 5 min followed by being immersed in 1% acid ethanol (1% HCl in 70% ethanol) for 10 s and then rinsed in water for 1 h. Finally, the sections were stained with eosin solution for 3 min and followed by dehydration with graded alcohol and clearing in xylene [14 (link)]. The slides were examined and photographed using an Olympus BX61 fluorescence microscope (Japan). Observed morphological structure of cells. The size of cells (4 random fields of equal area) was analyzed by Image-Pro Plus 6.0 software.
10
Histological Analysis of Mouse Eyeballs
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The mice were sacrificed at 3 d p.i. and 6 d p.i.. Eyeballs were immersed in 4% paraformaldehyde for 24 h to be fixed and embedded in paraffin after dehydration with an ethanol gradient (70, 80, 90, 95, and 100%). The eyeballs tissue was sliced into 3-μM-thick sections, deparaffinized with xylene, and then hydrated in an ethanol gradient. Then, the sections were stained with hematoxylin-eosin staining kit (Servicebio, Wuhan, China). Photos were taken of the sections using a light microscope (Leica Microsystems, Wetzlar, Germany).
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