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Internet-Based Image Analysis Quantifies Contractile Behavior of Individual Fibroblasts inside Model Tissue

Steven Vanni^*, B. Christoffer Lagerholm^*, Carol Otey, D. Lansing Taylor^* and Frederick Lanni^*

^* Department of Biological Sciences, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania 15213; and Department of Cell and Molecular Physiology, University of North Carolina, Chapel Hill, North Carolina 27599

Correspondence: Address reprint requests to Frederick Lanni, Carnegie Mellon University, 4400 Fifth Ave., Box 32, Pittsburgh, PA 15213. Tel.: 412-268-3460; E-mail: lanni{at}cmu.edu.

In a cell-populated collagen gel, intrinsic fiber structure visible in differential interference contrast images can provide markers for an in situ strain gauge to quantify cell-gel mechanics, while optical sections of fluorescent protein distribution capture cytoskeletal kinematics. Mechanics quantification can be derived automatically from timelapse differential interference contrast images using a Deformation Quantification and Analysis software package accessible online at http://dqa.web.cmu.edu. In our studies, fibroblast contractile machinery was observed to function entirely within pseudopods, while GFP-alpha-actinin concentrated in pseudopod tips and cortex. Complex strain patterns around individual cells showed instances of both elastic and inelastic strain transmission, suggesting a role in observed long-range alignment of cells.

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