LV and visceral (retroperitoneal) fat tissue was fixed with ice-cold 4% paraformaldehyde for 48 h, embedded in paraffin and processed for histology as described previously.15 (link) In brief, transverse sections were stained either with hematoxylin–eosin for routine histological examination or with Azan-Mallory solution for evaluation of fibrosis.15 (link) To evaluate macrophage infiltration into the myocardium and adipose tissue, we performed immunostaining for the monocyte–macrophage marker CD68 with the paraffin-embedded sections.10 (link) All image analysis was performed with NIH Scion Image software (Scion, Frederick, MD, USA).17 (link)
Nih scion image software
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NIH Scion Image software is a digital image analysis tool used for processing, analyzing, and measuring images. It provides basic image processing capabilities such as contrast adjustment, cropping, and measurement tools, allowing users to extract quantitative data from digital images.
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Lab products found in correlation
4 protocols using nih scion image software
1
Histological Analysis of Cardiac and Adipose Tissue
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Cardiac Tissue Histological Assessment
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LV tissue was fixed in ice‐cold 4% paraformaldehyde for 48 to 72 hours, embedded in paraffin, and processed for histology as described.20 (link) Transverse sections (thickness, 3 μm) of the left ventricle were stained either with hematoxylin‐eosin for routine histological examination or with Azan‐Mallory solution for evaluation of the extent of fibrosis. Image analysis was performed with NIH Scion Image software (Scion Corp, Frederick, MD) in a blinded manner to the experimental status of the animals.
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Macrophage Infiltration in Cardiac and Adipose Tissue
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The LV and visceral (retroperitoneal) fat tissue were fixed in ice‐cold 4% paraformaldehyde for 48 h, embedded in paraffin and processed for histology, as described 18 , 19 . For evaluation of macrophage infiltration into the LV myocardium and adipose tissue, paraffin‐embedded tissue sections were subjected to immunohistochemical staining for the monocyte–macrophage marker CD68, as described previously 14 . All image analyses were performed with the use of nih scion image software (Scion, Frederick, MD, USA).
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Immunostaining for Macrophage Infiltration in LV Myocardium
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LV tissue was fixed in ice-cold 4% paraformaldehyde for 48 h, embedded in paraffin, and processed for histology, as described.7 ) To evaluate macrophage infiltration into the LV myocardium, we performed immunostaining for the monocyte-macrophage marker CD68 with frozen sections (thickness, 5 µm) that had been fixed with acetone. Endogenous peroxidase activity was blocked by exposure of the sections to methanol containing 0.3% hydrogen peroxide. Sections were incubated at 4°C first overnight with mouse monoclonal antibodies to CD68 (1:100 dilution of clone ED1; Chemicon, Temecula, CA, USA) and then for 30 min with Histofine Simple Stain Rat MAX PO (Nichirei Biosciences, Tokyo, Japan). Immune complexes were visualized with diaminobenzidine and hydrogen peroxide, and the sections were counterstained with hematoxylin. All image analysis was performed with NIH Scion Image software (Scion, Frederick, MD, USA).
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