The esophagus was fixed in 40 g/L formaldehyde saline solution, embedded in paraffin, and cut into 5-mm sections. Sections were made along the minor axis of the esophagus. Tissue sections underwent hematoxylin and eosin staining and immunostaining to assess the condition of the wound tissue. For the immunostaining, we used anti-α-smooth muscle actin (α-SMA) antibody (clone 1A4, 1:1000 dilution; Sigma-Aldrich), anti-calponin-1 (CNN1) antibody (1:1000 dilution; Abcam, Cambridge, UK), anti-myeloperoxidase (MPO) antibody (1:300 dilution; Thermo Scientific, Waltham, Mass), and anti-CD107a antibody (clone 4E9/11, 1:300 dilution; AbD Serotec, Kidlington, UK). We photographed five fields per section from each pig and measured the stained area per field of view (× 200, scale bar: 50 μm) with a digital image analyzer (WinROOF 2018; Mitani Co., Fukui, Japan). Moreover, collagen fibers were stained with Elastica–Masson staining. We randomly selected five locations, including the thickest and thinnest stained fiber layers, and measured the thickness using cellSens (Olympus, Tokyo, Japan) at × 40 (scale bar: 200 μm). To evaluate the state of tissue regeneration in the entire wound, we measured not only the fibrous layer but also the thickness from the bottom edge of the area stained with Elastica–Masson staining to the surface layer of the wound.
Anti myeloperoxidase mpo antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-myeloperoxidase (MPO) antibody is a laboratory reagent used in immunoassays and research applications. It is a specific antibody that binds to the myeloperoxidase protein, which is an enzyme found in the cytoplasm of neutrophils and monocytes. The antibody can be used to detect and quantify the presence of myeloperoxidase in biological samples.
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3 protocols using anti myeloperoxidase mpo antibody
1
Histopathological Assessment of Esophageal Wound Healing
2
Esophageal Histological Analysis
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The esophagus was fixed in 40 g/l of formaldehyde saline, embedded in paraffin and cut into 5 μm sections. Tissue sections were stained with Masson's trichrome to examine the accumulation of collagen fibers. Three fields on a section from each pig were photographed, and the thickness of the stained areas was measured (200) with a digital image analyser (WinROOF, Mitani Co., Fukui, Japan and NDP. view2 software, Hamamatsu, Japan).
The tissue sections were stained with anti-α-smooth muscle actin (α-SMA) antibody (clone 1A4, dilution; 1:1,000, Sigma-Aldrich) for 60 min, anti-myeloperoxidase (MPO) antibody (dilution; 1:300, Thermo Scientific, Waltham, Massachusets, USA) for 40 min, anti-CD107a antibody (clone 4E9/11, dilution; 1:300, AbD Serotec, Kidlington, UK) for 60 min, anti-CD31 antibody (clone M-20, dilution; 1:600, Santa Cruz Biotechnology, Dallas, Texas, USA) for 30 min and anti-Ki-67 antibody (clone MIB-1, dilution; 1:100, Dako, Glostrup, Denmark) for 60 min at room temperature. Nine random fields on a section from each esophagus were photographed, and stained areas were calculated from the entire cross-sectional area of the esophagus.
The tissue sections were stained with anti-α-smooth muscle actin (α-SMA) antibody (clone 1A4, dilution; 1:1,000, Sigma-Aldrich) for 60 min, anti-myeloperoxidase (MPO) antibody (dilution; 1:300, Thermo Scientific, Waltham, Massachusets, USA) for 40 min, anti-CD107a antibody (clone 4E9/11, dilution; 1:300, AbD Serotec, Kidlington, UK) for 60 min, anti-CD31 antibody (clone M-20, dilution; 1:600, Santa Cruz Biotechnology, Dallas, Texas, USA) for 30 min and anti-Ki-67 antibody (clone MIB-1, dilution; 1:100, Dako, Glostrup, Denmark) for 60 min at room temperature. Nine random fields on a section from each esophagus were photographed, and stained areas were calculated from the entire cross-sectional area of the esophagus.
3
Histological Analysis of Post-ESD Rectum
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The rectum was fixed in 40 g/L of formaldehyde saline, embedded in paraffin and cut into 5-µm sections. Tissue sections were stained with hematoxylin and eosin (H&E), and Masson's trichrome staining was performed to examine the accumulation of collagen fibers. Three fields that vertically crossed the short axis of the ESD site from each pig were randomly selected and photographed. A line was drawn between both ends of the muscularis mucosae and a vertical line was drawn from the middle of this line (Fig. 1D). The thickness of each layer from the post-ESD pigs as well as the normal pigs (n = 3) was measured (NDP View2 software; Hamamatsu, Japan) in a blinded manner.
The tissue sections were stained with anti-α-smooth muscle actin (α-SMA) antibody Endoscopy (Sigma-Aldrich, St. Louis, Missouri, USA), a marker for myofibroblasts, anti-CD107a antibody (AbD Serotec, Kidlington, UK), a marker for macrophages, anti-CD31 antibody (Santa Cruz Biotechnology, Dallas, Texas, USA), a marker for endothelial cells, and with anti-myeloperoxidase (MPO) antibody (Thermo Scientific, Waltham, Massachusetts, USA), a marker for neutrophils. Nine random submucosa fields from each pig were photographed (high-powered fields (HPF), ×400) and the stained cells were counted in a blinded manner.
The tissue sections were stained with anti-α-smooth muscle actin (α-SMA) antibody Endoscopy (Sigma-Aldrich, St. Louis, Missouri, USA), a marker for myofibroblasts, anti-CD107a antibody (AbD Serotec, Kidlington, UK), a marker for macrophages, anti-CD31 antibody (Santa Cruz Biotechnology, Dallas, Texas, USA), a marker for endothelial cells, and with anti-myeloperoxidase (MPO) antibody (Thermo Scientific, Waltham, Massachusetts, USA), a marker for neutrophils. Nine random submucosa fields from each pig were photographed (high-powered fields (HPF), ×400) and the stained cells were counted in a blinded manner.
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