Isolation of Single Muscle Fibers in Preparation for Loading into Microfluidic Devices

Abstract

Modeling and simulation of microfluidic devices to capture single muscle fibers has led to several design possibilities. Before testing the fabricated devices, however, a procedure for the isolation of single muscle fibers from muscle tissue must be developed. The aim is to optimize a collagenase treatment that can yield consistent fibers of large quantity and dimensions that correspond to device tapering regions. Optimization of collagenase concentration and trituration showed that a 0.6% (w/v) solution produced the most fibers and the wide bore pipette tip method of trituration produced the longest fibers. Further analysis of centrifugation reinforced the theory of fiber separation and clean up based upon length. Future work includes testing of a larger sample size of muscle tissue/type, testing fiber functionality, preparing new methods of collagenase preparation and treatment, and seeing fiber response/analysis in the fabricated devices.