Zen black image processing software

Aortic root paraffin sections were deparaffinized and heat-mediated antigen retrieval was performed using Tris/EDTA (10 mM Tris, 1 mM EDTA, 0.05% Tween, pH 9.0). Sections were blocked for 1 hour at RT with 2% BSA in PBS, followed by incubation with anti-αSMA-Alexa Fluor 647 primary antibody at 4°C. After extensive washing in PBS + 0.1% Tween, sections were counterstained for 5 min with 1μg/mL DAPI (Sigma) and mounted with AquaMount (Polysciences, Inc.). Samples were visualized using a Zeiss AX10 confocal microscope. Fibrous cap smooth muscle cells were counted by positive αSMA staining and normalized to fibrous cap area measured using Zen Black image processing software (Zeiss).

Aortic root paraffin sections were deparaffinized and heat-mediated antigen retrieval was performed using Tris/EDTA (10 mM Tris, 1 mM EDTA, 0.05% Tween, pH 9.0). Sections were blocked for 1 h at RT with 2% BSA in PBS, followed by incubation with anti-αSMA-Alexa Fluor 647 primary antibody at 4°C. After extensive washing in PBS + 0.1% Tween, sections were counterstained for 5 min with 1 µg/ml DAPI (Sigma) and mounted with AquaMount (Polysciences, Inc.). Samples were visualized using a Zeiss AX10 confocal microscope. Fibrous cap smooth muscle cells were counted by positive α-SMA staining and normalized to fibrous cap area measured using Zen Black image processing software (Zeiss).