Single fiber dissociation of intact rodent muscles: Difference between revisions
This protocol is for isolating single muscle fibers from intact mouse skeletal muscles. It is optimized for use with the Flexor Digitorum Brevis (superficial foot) muscle. |
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##Optional additional cleaning step: Transfer fibers in incubation medium to 5 mL tubes & allow to settle via gravity sedimentation (approx. 20 min). Carefully remove medium & replace with fresh incubation medium. Repeat once more, before placing cleaned fibers into fresh culture dishes. | ##Optional additional cleaning step: Transfer fibers in incubation medium to 5 mL tubes & allow to settle via gravity sedimentation (approx. 20 min). Carefully remove medium & replace with fresh incubation medium. Repeat once more, before placing cleaned fibers into fresh culture dishes. | ||
#Let muscle fibers recover overnight. They are now ready for use experimentally. | #Let muscle fibers recover overnight. They are now ready for use experimentally. | ||
==References== | |||
This protocol is a modified method of: | |||
*Cho et al., (2016) FASEB J 30(2):674-87 http://www.fasebj.org/content/30/2/674.full | |||
*Spangenburg et al. http://www.seahorsebio.com/resources/tech-writing/techbrief-intact-muscle-fiber.pdf | |||