Generation of Porcine Ventricular Slices: A Vibratome-based Technique to Prepare Ultra-thin Slices of Pig Heart Tissue
Transcrição
Mammalian cardiac tissue slices – ultra-thin slices of viable mammalian ventricle heart tissue – retain the architecture and physiology of cardiac tissue, making them model systems for translational cardiovascular research.
To generate porcine cardiac tissue slices, place a freshly harvested porcine heart in a tray containing chilled cardioplegic solution, which temporarily ceases cardiac activity, protecting the heart. Dissect the heart to isolate the left ventricle, followed by cutting it into uniformly-sized tissue blocks. Using tissue glue, secure the tissue block flat to the premade agar block fixed to the base of the vibratome specimen holder, with the cardiac epicardium – the heart's outer layer – facing down.
Transfer the specimen holder to the chilled, oxygenated, buffer-filled, vibratome slicing tray to maintain the tissue's physiological conditions, and cool it during slicing. Adjust the vibratome blade height and cutting parameters based on the desired tissue section thickness. During the run, the vibrating blade advances and penetrates the tissue block.
The agar provides stability to the tissue block during slicing and maintains the blade on one path, generating uniform intact cardiac tissue slices with aligned myocardial fibers. Collect and transfer the tissue slices to an oxygenated buffer bath to reduce tissue damage. The cardiac tissue slices are ready for downstream analysis.
