Histological Evaluation of Myocardial Tissue

Hearts were fixed with 20% formalin (Sakura Finetek Japan Inc.) and embedded in paraffin. After cutting 4 µm each, paraffin sections were stained with anti-CD45 (BD Biosciences, USA), anti-F4/80 antibodies (Bio-Rad, USA), anti-iNOS antibody (abcam, UK), and anti-CD163 antibody (abcam) to evaluate cell infiltration. Elastica van Gieson staining and wheat germ agglutinin staining were performed to examine fibrosis proportion and calculate cell area, respectively. Cross-sectional area (CSA) of myocytes with a centrally located nucleus and overall circular shape was measured in the left ventricular free walls using Image J (NIH, USA)^{27 (link)}.

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Toko H., Morita H., Katakura M., Hashimoto M., Ko T., Bujo S., Adachi Y., Ueda K., Murakami H., Ishizuka M., Guo J., Zhao C., Fujiwara T., Hara H., Takeda N., Takimoto E., Shido O., Harada M, & Komuro I. (2020). Omega-3 fatty acid prevents the development of heart failure by changing fatty acid composition in the heart. Scientific Reports, 10, 15553.

Publication 2020

formalin anti-CD45 anti-F4/80 antibodies anti-iNOS antibody anti-CD163 antibody

Anti antibodies Anti antibody Cd163 Cell Elastica Embedded paraffin Fibrosis Formalin Hearts Inos Left ventricular Myocytes Nucleus Paraffin Wheat germ agglutinin

Corresponding Organization : The University of Tokyo

Other organizations : Shimane University

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Variable analysis

independent variables

20% formalin (Sakura Finetek Japan Inc.) for tissue fixation
Paraffin embedding of samples

dependent variables

Cell infiltration evaluated by staining with anti-CD45, anti-F4/80, anti-iNOS, and anti-CD163 antibodies
Fibrosis proportion examined by Elastica van Gieson staining
Cell area calculated by wheat germ agglutinin staining
Cross-sectional area (CSA) of myocytes with a centrally located nucleus and overall circular shape in the left ventricular free walls

control variables

Cutting paraffin sections at 4 µm thickness

controls

No positive or negative controls were explicitly mentioned in the provided information.

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