To begin, position the freeze dried muscle sample under the stereo microscope and view it at low magnification. With one pair of fine tissue dissecting forceps, hold the muscle in place and separate the small bundles of fibers with the other forceps. Isolate one bundle of fibers and gently tease them apart until single fiber segments are extracted from the bundle.
Then shift the fiber to a clear area of the Petri dish. Investigate each single fiber segment under 50 times magnification. Attempt to further separate the fiber at one end.
If the fiber begins to break, it is a single fiber. Carefully collect the fiber with a pair of forceps and place it into the previously prepared aliquot of denaturing buffer. Tap the tube firmly on the bench three times.
Afterward, vortex the fiber samples and briefly centrifuge them for five seconds at 2, 500G to ensure the samples settles at the bottom of the tube. Let the samples rest at room temperature for one hour before storing them at minus 80 degrees Celsius for future use.
