Dm5500 b 11888817 12 microscope

For quantitative evaluation, three H&E- and Masson-stained sections per rat were used. By applying ImageJ software (NIH, Bethesda, MD, USA), the H&E- and Masson-stained sections were examined to determine the cardiomyocyte area and Masson’s-stained area, respectively. A Leica DMLB2/11888111 microscope equipped with a Leica DFC450 camera was used to acquire the images.
Connexin-43 immunofluorescence was measured by randomly capturing five non-overlapping images per slide. A Leica DM5500 B/11888817/12 microscope, fitted with a Leica HI PLAN 10×/0.25 objective and a Leica DFC450C camera, was used to capture the images. Connexin-43-stained spots were manually counted using the plugin/cell counting tool (Rangan & Tesch, 2007 (link)) in ImageJ software (National Institutes of Health, Bethesda, MD, USA), and the average per field for each rat was then calculated. This measurement was performed in a blind manner by an independent experienced researcher. For statistical analysis and comparison, ten animals were used per experimental group.

For the quantitative assessment of immunostaining, three nonoverlapping images were randomly captured per section using a Leica DML B2/11888111 microscope equipped with a Leica DFC450 camera and Leica C PLAN 10 × 0.22 objective or a Leica DM5500 B/11888817/12 microscope equipped with a Leica DFC450C camera and Leica HI PLAN 10 × 0.25 objective. For each image, the field of view was at 10 × magnification. The number of immunopositive cells in the fields from at least three sections/animal was counted using ImageJ software (US National Institutes of Health, Bethesda, MD, USA) and averaged per field for each animal. This was blindly performed by an independent, experienced researcher. The calculated percentages for at least eight animals per experimental group were considered for comparison and statistical analyses.