To begin, embed the lung tissue in agarose. To prepare the lung tissue for sectioning, first, attach each tissue block to the specimen disc of the vibratome with super glue and add ice to the surrounding bath. Then carefully install the specimen disc into the buffer tray and fill the tray with PBS.
Next, set the vibratome parameters, keeping the thickness at 200 micrometers, the frequency at 100 hertz, the amplitude of the blade at 1.3 millimeters, and the forward speed of the blade between 0.02 and 0.03 millimeters per second. Commence the sectioning of the lung tissue. Once enough slices have been created, use a brush to gently lift the tissue slice from the vibratome tray.
Transfer the slices to a 24 well plate with PBS azide solution. Preserve the lung slices at four degrees Celsius.
