Isolation of Single Fibers for Seahorse Assays: Difference between revisions
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==Muscle Fiber Dissection and Digestion== | ==Muscle Fiber Dissection and Digestion== | ||
* Place ~20 mL KRBH, ~20 mL DMEM and ~100 mL aCSF Media in a 37C water bath to warm. | * Place ~20 mL KRBH, ~20 mL DMEM and ~100 mL aCSF Media in a 37C water bath to warm. | ||
* | * Sterilize surgical tools for ~30 min and move mice to the procedure room. | ||
* Dissect FDB muscles from mice (see [[FDB Muscle Dissection from Mice]]). Place both muscles from a mouse in a single 12-well containing 1 mL of pre-warmed KRBH. | * Dissect FDB muscles from mice (see [[FDB Muscle Dissection from Mice]]). Place both muscles from a mouse in a single 12-well containing 1 mL of pre-warmed KRBH. | ||
* Weigh out enough collagenase to make 1 mL of a 4 mg/mL solution for each mouse. Add DMEM to make the 4 mg/mL solution | * Weigh out enough collagenase to make 1 mL of a 4 mg/mL solution for each mouse. Add DMEM to make the 4 mg/mL solution | ||