To begin, inspect the muscles of interest dissected from the sacrificed mouse for the absence of bone chips or ragged edges and trim them away with sharp scissors if needed. Weigh the cleaned muscles using an analytical balance and record their weight. After placing the muscle in at least 0.1 milliliters of 1%SDS per milligram of weight, place the well plate on a rocking shaker set to 50 to 80 Hertz.
Visually inspect the SDS solution periodically. When the solution becomes cloudy, remove the solution with a pipette without aspirating the muscles and replace it with an equal volume of fresh 1%SDS. When the solution remains clear for 24 hours, remove the final SDS solution from the decellularized muscles and replace it with an equal volume of PBS.
Inspect the decellularized muscles under a stereo microscope and carefully remove any hair or debris stuck to the muscle using forceps. Carefully remove the PBS and replace it with an equal volume of 3.7%formaldehyde before returning the plate to the rocking shaker for 24 hours.
