Deformation Tracking in PAA Gels using TFM

For tracking deformation in PAA gels^{11 (link)} during TFM analysis, we modified patterned PAA platforms by adding 0.2 µm fluorescent nanobeads (Polysciences, Cat#BLI832-1) into PAA solutions before gel polymerization. Before and after removal of cells, images of the PAA micropatterns were taken with live confocal laser scanning at the interface planes between cells and the adhesion ligand patterns. Live cell imaging was performed in a microclimate-controlled stage top incubator (Tokai Hit, Japan) at 37 °C in 5% CO₂. Bead displacements and corresponding traction forces fields were calculated using an iterative particle image velocimetry algorithm and an unconstrained Fourier transform traction cytometry algorithm, respectively (ImageJ plugins).

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Tabdanov E.D., Puram V.V., Win Z., Alamgir A., Alford P.W, & Provenzano P.P. (2018). Bimodal sensing of guidance cues in mechanically distinct microenvironments. Nature Communications, 9, 4891.

Publication 2018

microclimate-controlled stage top incubator

Cat 1 Cell Displacements Ligand Microclimate Polymerization Traction Velocimetry

Corresponding Organization :

Other organizations : University of Minnesota

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Variable analysis

independent variables

Modification of patterned PAA platforms by adding 0.2 µm fluorescent nanobeads (Polysciences, Cat#BLI832-1) into PAA solutions before gel polymerization

dependent variables

Bead displacements and corresponding traction forces fields

control variables

Live cell imaging performed in a microclimate-controlled stage top incubator (Tokai Hit, Japan) at 37 °C in 5% CO2

controls

Before and after removal of cells, images of the PAA micropatterns were taken with live confocal laser scanning at the interface planes between cells and the adhesion ligand patterns.

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