Quantifying Myotube Contractility Dynamics

After 14 days in differentiation medium, mutant and WT myotubes cultures in petri dishes were placed on a 37 °C heated stage (ThorLabs TC200) on an upright phase-contrast microscope (Zeiss Hal 100). Myotube contraction was elicited using silver electrode-driven, 1 V broad field stimulation at frequencies of 0.3, 0.5, 1.0, 2.0 and 4.0 Hz for 15 s. Myotube contractions were captured using a high-speed acquisition digital camera (Hamamatsu model C8484-05G) from a selected region of interest (ROI). This procedure is described in detail by Santhanam et al.^{57 (link)}. Contraction synchrony was calculated by dividing the number of contractions by the total number of expected contractions at a given frequency.

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Badu-Mensah A., Guo X., McAleer C.W., Rumsey J.W, & Hickman J.J. (2020). Functional skeletal muscle model derived from SOD1-mutant ALS patient iPSCs recapitulates hallmarks of disease progression. Scientific Reports, 10, 14302.

Publication 2020

TC200 Hal 100

Dishes High Myotube Phase contrast microscope Silver

Corresponding Organization :

Other organizations : University of Central Florida, Hesperos (United States)

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Variable analysis

independent variables

Stimulation frequency (0.3, 0.5, 1.0, 2.0, 4.0 Hz)

dependent variables

Myotube contraction
Contraction synchrony

control variables

Mutant and WT myotube cultures
Petri dish culture conditions
37°C heated stage
Upright phase-contrast microscope
Silver electrode-driven, 1 V broad field stimulation
High-speed acquisition digital camera

controls

Mutant and WT myotube cultures

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