Improved-throughput traction microscopy based on fluorescence micropattern for manual microscopy

PLoS One. 2013 Aug 1;8(8):e70122. doi: 10.1371/journal.pone.0070122. Print 2013.

Abstract

Traction force microscopy (TFM) is a quantitative technique for measuring cellular traction force, which is important in understanding cellular mechanotransduction processes. Traditional TFM has a significant limitation in that it has a low measurement throughput, commonly one per TFM dish, due to a lack of cell position information. To obtain enough cellular traction force data, an onerous workload is required including numerous TFM dish preparations and heavy cell-seeding activities, creating further difficulty in achieving identical experimental conditions among batches. In this paper, we present an improved-throughput TFM method using the well-developed microcontact printing technique and chemical modifications of linking microbeads to the gel surface to address these limitations. Chemically linking the microbeads to the gel surface has no significant influence on cell proliferation, morphology, cytoskeleton, and adhesion. Multiple pairs of force loaded and null force fluorescence images can be easily acquired by means of manual microscope with the aid of a fluorescence micropattern made by microcontact printing. Furthermore, keeping the micropattern separate from cells by using gels effectively eliminates the potential negative effect of the micropattern on the cells. This novel design greatly improves the analysis throughput of traditional TFM from one to at least twenty cells per petri dish without losing unique advantages, including a high spatial resolution of traction measurements. This newly developed method will boost the investigation of cell-matrix mechanical interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • HeLa Cells
  • Humans
  • Mechanical Phenomena*
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Microspheres
  • Surface Properties

Grants and funding

The work reported here was supported by the National Basic Research Program of China (Grant Nos. 2013CB933702 and 2011CB809106) and the National Natural Science Foundation of China (Grant Nos.11002003 and 11072004). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.