Quantification of Cardiac Fibrosis

Hearts were removed and fixed in 10% formalin for 48 hours, dehydrated with ethanol, and embedded in paraffin. Sections were cut at 7 µm, with 50 µm between each section, and Masson’s trichrome staining was performed (Sigma-Aldrich, HT15-1KT) according to the manufacturer’s instructions. Analysis and quantification of fibrosis was performed as previously described^{46 (link),50 (link)}, where 3 sections per heart were imaged on an Olympus BX51 microscope and quantified using ImageJ (NIH, Bethesda, MD).

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Ghnenis A.B., Burns D.T., Osimanjiang W., He G, & Bushman J.S. (2020). A Long-Term Pilot Study on Sex and Spinal Cord Injury Shows Sexual Dimorphism in Functional Recovery and Cardio-Metabolic Responses. Scientific Reports, 10, 2762.

Publication 2020

HT15-1KT

Dehydrated ethanol Embedded paraffin Fibrosis Formalin Heart Microscope

Corresponding Organization :

Other organizations : University of Wyoming

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Variable analysis

independent variables

Removal and fixation of hearts in 10% formalin for 48 hours
Dehydration of hearts with ethanol
Embedding of hearts in paraffin

dependent variables

Fibrosis quantification

control variables

Sections cut at 7 µm with 50 µm between each section
Masson's trichrome staining performed according to manufacturer's instructions
Imaging of 3 sections per heart on an Olympus BX51 microscope
Quantification of fibrosis using ImageJ (NIH, Bethesda, MD)

controls

Positive control not specified
Negative control not specified

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