Azan staining of the specimens was performed at one week after the operation. The area and thickness of the newly formed granulation tissue on the sections prepared from the center of the wounds was measured. The granulation thickness was measured at three points (left edge, center and right edge of the newly formed granulation tissue in the wound) in each section and the mean thickness was used. The granulation area was measured according to the previously reported method [20] . Briefly, the area between both levels of marginal skin on the underlying muscle layer was measured on Azan-stained sections using an optical microscope.(n=5 in each width and height were chosen from the central region beyond the muscle layer. The number and cross-sectioned area of newly formed capillaries in the two squares in each section were measured using the BZ-II Analyzer imaging software program (version 1.42; KEYENCE Co., Osaka, Japan), and the mean values were calculated.
Bz 2 analyzer imaging software program
Manufactured by Keyence
Sourced in Japan
The BZ-II Analyzer is an imaging software program developed by Keyence. It is designed to capture and analyze high-resolution images. The software provides tools for image processing, measurement, and analysis, but does not include any interpretation or extrapolation of the data.
Automatically generated - may contain errors
2 protocols using bz 2 analyzer imaging software program
1
Granulation Tissue Analysis in Wound Healing
2
Quantifying Angiogenesis via vWF Immunohistochemistry
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Immunohistochemical staining with von Willebrand factor (vWF) was used to detect newly formed capillaries. For this, 6-μm sections were dewaxed, rehydrated, and incubated with proteinase K (S3020; Dako Japan, Tokyo, Japan) for 5 minutes at room temperature for antigen retrieval. Anti-vWF rabbit polyclonal antibodies (1:5000, Code No. A0082; Dako) were used as the primary antibody followed by the secondary antibody (K4003, EnVision; Dako). The staining was visualized using 3-3’-diaminobenzidine-4HCl (DAB, Code 725191; Nichirei Biosciences Inc., Tokyo, Japan); the sections were then counterstained with hematoxylin and micrographs were taken under an optical microscope.
In each section, a rectangle of 500 × 300 μm was selected at the center of the granulation area beyond the muscle layer, and the number of newly formed capillaries in the rectangle and the cross-sectioned area of neovascularization were measured using the BZ-II Analyzer imaging software program (version 1.42; KEYENCE Co.).
In each section, a rectangle of 500 × 300 μm was selected at the center of the granulation area beyond the muscle layer, and the number of newly formed capillaries in the rectangle and the cross-sectioned area of neovascularization were measured using the BZ-II Analyzer imaging software program (version 1.42; KEYENCE Co.).
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