High Throughput Traction Force Microscopy for Multicellular Islands on Combinatorial Microarrays

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Abstract

The composition and mechanical properties of the cellular microenvironment along with the resulting distribution of cellular devolved forces can affect cellular function and behavior. Traction Force Microscopy (TFM) provides a method to measure the forces applied to a surface by adherent cells. Numerous TFM systems have been described in literature. Broadly, these involve culturing cells on a flexible substrate with embedded fluorescent markers which are imaged before and after relaxion of cell forces. From these images, a displacement field is calculated, and from the displacement field, a traction field. Here we describe a TFM system using polyacrylamide substrates and a microarray spotter to fabricate arrays of multicellular islands on various combinations of extra cellular matrix (ECM) proteins or other biomolecules. A microscope with an automated stage is used to image each of the cellular islands before and after lysing cells with a detergent. These images are analyzed in a semi-automated fashion using a series of MATLAB scripts which produce the displacement and traction fields, and summary data. By combining microarrays with a semi-automated implementation of TFM analysis, this protocol enables evaluation of the impact of substrate stiffness, matrix composition, and tissue geometry on cellular mechanical behavior in high throughput.

Original languageEnglish (US)
JournalBio-protocol
Volume9
Issue number21
DOIs
StatePublished - Nov 5 2019

Keywords

  • Traction force microscopy
  • Cellular forces
  • Mechanobiology
  • Microenvironment
  • Substrate stiffness
  • Microarray
  • Multicellular island
  • TFM

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