Traction Assays for Studies of Cell Mechanotransduction V. Damljanović 1 , B. Lagerholm 1, M. Dembo 2 & K. Jacobson 1 1 Cell & Developmental Biology, University of North Carolina, Chapel Hill, NC; 2 Biomedical Engineering, Boston University, Boston, MA
Traction Assays for Studies of Cell Mechanotransduction V. Damljanovi ć 1 , B. Lagerholm 1, M. Dembo 2 & K. Jacobson 1 1 Cell & Developmental Biology, University of North Carolina, Chapel Hill, NC; 2 Biomedical Engineering, Boston University, Boston, MA. Cell Mechanotransduction. - PowerPoint PPT Presentation
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Traction Assays for Studies of Cell Mechanotransduction
V. Damljanović1, B. Lagerholm1, M. Dembo2 & K. Jacobson1
1Cell & Developmental Biology, University of North Carolina, Chapel Hill, NC; 2Biomedical Engineering, Boston University, Boston, MA
Cell Mechanotransduction
Sense environment
Correlate w/ cell state
Feedback
Signal Cytoskeleton
Apply tractions
Motion
Cell Tractions
Tractions are determined from the deformation of substrate
Adhesion moleculesElastic
substrate
“Frictional” tractions
“Propulsive” tractions
Direction of migration
Substrate tractions
Polyacrylamide gel on 22 x 22 mm coverslip (modified protocol of Yu-li Wang)
Experimental Requirements(must match theoretical assumptions)
• Gel must be flat, with free edges and bottom fixed on the coverslip
• Gel thickness must be orders of magnitude greater than average displacement, but small enough for optics (70-100m optimal)
• Fluorescent markers must be small (we use 0.2m) and only at the top (not really the case)
• Must keep the focus always at the same set of beads (difficult)
• Must be isolated from all vibrations—translation is tolerable, but not rotation