2.5. Confocal Microscopy

Paraffin embedded 0.5 µm heart sections were incubated with anti-MMP-9 (diluted 1:500 in PBS 1.5% BSA) primary antibody overnight at 4 °C. After washing three times with PBS, the slides were incubated with Alexa-Fluor-647 conjugated secondary antibody for 1 h at room temperature. Slides were washed three times with PBS and mounted in PBS media, containing Hoechst for nuclei visualization. Images were taken using a Leica TCS SP5 confocal microscope. At least three different fields per condition were obtained.