Lung tissues were harvested for observing morphologic alterations at 24 h after CLP/sham modelling. The right lung lobes were dissected, washed and fixed with 4% (v/v) paraformaldehyde for 24 h at 4 °C. Lung tissues were embedded in paraffin, sectioned at 4 μm thickness, dewaxed and rehydrated and stained with hematoxylin and eosin (H&E) solution (hematoxylin, MHS16; eosin, HT110132; Sigma-Aldrich, USA) for histological examination. The stained slides were then observed with the light microscope and the digital micrographs were taken for analyzing. Histologic changes were evaluated by two independent pathologists blinded to the experiment. The histologic injury scores [21 (link)] were calculated according to the sum of the score for alveolar edema, alveolar hemorrhage, pulmonary interstitial thickening and neutrophil infiltration. Each histological characteristic was evaluated on a scale from 0 to 3: 0, normal (no injury); 1, minimal (injury to 25% of the field); 2, mild (injury between 25 and 50% of the field); 3, moderate (injury between 50 and 75% of the field); 4, severe (injury over 75% of the field).
Ht110132
Manufactured by Merck Group
Sourced in United States, Japan
The HT110132 is a piece of lab equipment manufactured by Merck Group. It is designed for specific laboratory tasks, but a detailed and unbiased description of its core function cannot be provided while maintaining conciseness and avoiding interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Mhs16, by Merck Group (9 mentions)
Mounting medium, by Thermo Fisher Scientific (4 mentions)
Intensilight c hgfi, by Nikon (2 mentions)
Hhs32, by Merck Group (2 mentions)
Image analyzer, by Molecular Devices (1 mentions)
Anti ki67 ab16667, by Abcam (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
Dab peroxidase substrate kit, by Vector Laboratories (1 mentions)
Sh30 500d, by Thermo Fisher Scientific (1 mentions)
Haematoxylin, by Merck Group (1 mentions)
Oil red o, by Merck Group (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Microtome, by Leica (1 mentions)
Cx31 light microscope, by Olympus (1 mentions)
Rm2245, by Leica (1 mentions)
19 protocols using ht110132
1
Histopathological Lung Assessment in Sepsis
2
Histological Analysis of Muscle Fiber CSA
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GA muscles were immersed in an optimal cutting temperature (OCT) solution immediately after dissection and frozen at −80 °C. OCT blocks were cut to a thickness of 10 μm and stained with hematoxylin (30002; Muto Pure Chemicals Co., Ltd., Tokyo, Japan) and eosin (H&E, HT110132; Sigma-Aldrich). Subsequently, these stained sections were examined (200× magnification) for cross-sectional area (CSA) analysis under a confocal microscope (Nikon Intensilight C-HGFI, Tokyo, Japan) and NIS-element AR 4.00.00 software. The CSA of myofibers was then measured using the ImageJ software.
3
Histological Analysis of Prostatic Tissue
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Fixed prostatic tissue embedded in paraffin wax was cut into 4 μm thick sections and stained with hematoxylin (MHS-16; Sigma-Aldrich) and eosin (HT110–1-32; Sigma-Aldrich). Coverslips were mounted on sections using mounting medium (Invitrogen, Carlsbad, CA, USA) and then the sections were examined under a microscope (Nikon, Tokyo, Japan). Prostatic epithelial thickness was measured using an image analyzer (Molecular Devices Inc., CA, USA) as described previously [13 (link)].
4
Histological Analysis of Prostate Tissues
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Fixed prostate tissues were embedded in paraffin wax and cut into 5 μm thick sections. The sections were deparaffinized, rehydrated, and stained with Mayer's hematoxylin (MHS-16, Sigma-Aldrich) and eosin (HT110-1-32, Sigma-Aldrich) (H&E) solution using standard procedures. The sections were mounted with mounting medium (Invitrogen, Carlsbad, CA) and observed under light microscopy with bright-field illumination (Olympus, Tokyo, Japan).
Immunohistochemistry was performed using a Vectastain Elite ABC Kit (Vector Laboratories Inc., Burlingame, CA) according to the manufacturer's instructions. After antigen retrieval processing, the sections were blocked in normal serum and then incubated with anti-Ki-67 (ab16667, Abcam, Cambridge, UK) antibody overnight at 4°C. On the next day, the sections were incubated in biotinylated-secondary antibody and developed using a DAB peroxidase substrate kit (Vector Laboratories) until a signal was seen.
Immunohistochemistry was performed using a Vectastain Elite ABC Kit (Vector Laboratories Inc., Burlingame, CA) according to the manufacturer's instructions. After antigen retrieval processing, the sections were blocked in normal serum and then incubated with anti-Ki-67 (ab16667, Abcam, Cambridge, UK) antibody overnight at 4°C. On the next day, the sections were incubated in biotinylated-secondary antibody and developed using a DAB peroxidase substrate kit (Vector Laboratories) until a signal was seen.
5
Histological Evaluation of Muscle Fibers
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GA muscles were fixed in 10% neutral buffered formalin and embedded in paraffin. These paraffin blocks were cut into 5-μm-thick sections and stained with hematoxylin (30002, MUTO PURE CHEMICALS CO., LTD., Japan) and eosin (HT110132, Sigma, USA) (H&E). The H&E-stained sections were used for CSA analyses and examined (200× magnification) under a confocal microscope (Nikon Intensilight C-HGFI, Japan) using NIS-element AR 4.00.00 software. The myofiber CSAs were analyzed with ImageJ program.
6
Histological Analysis of Muscle Fiber
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TA muscles were fixed in 10% neutral buffered formalin and embedded in paraffin. These paraffin blocks were cut into 4 µm thick sections and stained with hematoxylin (30002, Muto Pure Chemicals Co., Ltd., Japan) and eosin (HT110132, Sigma-Aldrich, MO, USA) (H&E). The H&E-stained sections were used for the cross-sectional area (CSA) analyses. These sections were examined (200 × magnification) using a confocal microscope (Nikon intensilight C-HGFI, Tokyo, Japan) and NIS-element AR 4.00.00 software. Then, the myofiber cross-sectional areas were analyzed using the ImageJ software.
7
Histological Staining of Muscle Tissue
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H&E staining and Oil Red O staining were performed according to standard protocols. For H&E staining, the muscle sections were fixed in 4% PFA and incubated in haematoxylin (Fisher Scientific, SH30-500D) for 15 min. After washing, the sections were differentiated in 1% acid alcohol and saturated lithium carbonate solution, followed by five dips in eosin (Sigma, HT-110132) and extensive washing. Next, the sections were dehydrated in gradient alcohol and xylene and mounted in DPX. For Oil Red O staining, the sections were fixed in PFA, washed in PBS and rinsed in 60% isopropanol. After incubating in Oil Red O (Sigma, O1391) working solution for 15 min, the sections were rinsed in 60% isopropanol and stained in haematoxylin for 2 min. Then, the sections were washed in water and mounted in VECTASHIELD mounting medium (Vector Lab, H-1,000).
8
Histological Analysis of Prostate Tissues
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Fixed prostate tissues embedded in paraffin wax were cut into 4 μm thick sections and stained with hematoxylin (Sigma-Aldrich, MHS-16) and eosin (Sigma-Aldrich, HT110-1-32). The sections were mounted and cover-slipped using mounting medium (Invitrogen, Carlsbad, CA, USA) and then examined under a microscope (Zeiss, Oberkochen, Germany).
9
Histopathological Evaluation of Lung Injury
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Lung tissues were harvested for observing morphologic alterations at 24 h after LPS or PBS administration. The right middle lobe of lung were excised, washed and fixed with 4% (v/v) paraformaldehyde for 24 h at 4 °C. Lung tissues were embedded in paraffin, sectioned at 4 μm thickness, dewaxed and rehydrated, and stained with hematoxylin and eosin (H&E) solution (hematoxylin, MHS16; eosin, HT110132; Sigma-Aldrich, USA) to estimate inflammation in alveolar and peribronchial lesions. The stained slides were then observed with the light microscope and the digital micrographs were taken for analyzing. Histologic changes were evaluated by a pathologist blinded to the experiment. The degree of lung injury was graded using a histologic ALI scoring system based on histologic features, including neutrophils infiltration, hyaling membranes, proteinaceous debris, and alveolar septal thickening [16 (link)].
10
Evaluating Metastatic Lung Tumors
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Mouse lungs were resected out and fixed with 3.7% paraformaldehyde for the evaluation of metastasizing tumors. Tissues embedded in OCT compound (4583, Tissue-Tek, Torrance, CA, USA) were sectioned (9 μm), mounted, and then stained for 13 min with hematoxylin (HEM001, Biopure, Chungju, Republic of Korea) followed by eosin (HT110132, Sigma) for 5 min.
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