LV cross-sections (5μm) were stained with Wheat Germ Agglutinin (WGA, Cat#W11262, Thermo Fisher, Waltham, MA) to assess cardiomyocyte cross-sectional area [53 (link)] and with trichrome (Cat# 9179; Newcomer Supply, Middleton, WI) to assess fibrosis. Slides (one per animal) were examined and semi-automatically quantified using ZEN® (Carl Zeiss Microscopy, Cambridge, UK) [12 (link)]. Hematoxylin & eosin (H&E) staining was performed to assess capillary density [33 (link)]. In each slide, 10-15 high-magnification fields were selected randomly and results from all fields were averaged. To assess angiogenic signaling, myocardial expression of vascular endothelial growth factor (VEGF, Santa Cruz, Dallas, Texas, 1:200) was measured by western blot in homogenized LV myocardium.
Protocol detail
Cardiac Remodeling Histopathology Analysis
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Angiogenic
Animal
Capillary
Cardiomyocyte
Eosin
Fibrosis
Microscopy
Myocardium
Vegf
Western blot
Wheat germ agglutinin
Corresponding Organization :
Other organizations : Mayo Clinic, Mayo Clinic in Arizona
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Variable analysis
independent variables
- Wheat Germ Agglutinin (WGA) staining
- Trichrome staining
- Hematoxylin & eosin (H&E) staining
- Vascular endothelial growth factor (VEGF) expression measured by western blot
- Cardiomyocyte cross-sectional area
- Fibrosis
- Capillary density
- Angiogenic signaling (VEGF expression)
- Slide (one per animal)
- Positive control: Not specified
- Negative control: Not specified
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