Traction Force Microscopy images were acquired as previously described52 (link),63 (link). NorHA hydrogel films were prepared as described with 0.2 μm diameter fluorescent beads at 1% vol/vol (Invitrogen F8810). hMSCs were cultured for 18 hours before TFM analysis. Embedded beads and cells were captured in phase contrast and fluorescence using a DeltaVision Deconvolution Microscope (GE Healthcare Life Sciences, Marlborough, MA). Images were acquired prior and after cell lysis with PBS buffer containing 10% SDS (sodium dodecyl sulfate)/1% Triton X-100. ImageJ was used for TFM data analysis, including stack alignment, particle image velocimetry (PIV), Fourier transform traction cytometry (FTTC))64 (link). For FTTC variables, a Poisson’s ratio of 0.45 was assumed and 1e−9 was utilized as regularization parameter.
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Traction Force Microscopy of hMSCs
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Buffer
Cell
Fluorescence
Force microscopy
Hydrogel films
Microscope
Phase contrast
Sodium dodecyl sulfate
Traction
Triton x 100
Velocimetry
Corresponding Organization :
Other organizations : University of Pennsylvania
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Variable analysis
independent variables
- NorHA hydrogel film preparation (with 0.2 μm diameter fluorescent beads at 1% vol/vol)
- Traction forces exerted by hMSCs, measured using Traction Force Microscopy (TFM)
- HMSCs were cultured for 18 hours before TFM analysis
- Imaging of embedded beads and cells using phase contrast and fluorescence microscopy
- Acquisition of images before and after cell lysis with PBS buffer containing 10% SDS/1% Triton X-100
- Poisson's ratio of 0.45 and regularization parameter of 1e-9 used for Fourier transform traction cytometry (FTTC) analysis
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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