Abstract
The interactions between biochemical processes and mechanical signaling play important roles during various cellular processes such as wound healing, embryogenesis, metastasis, and cell migration. While traditional traction force measurements have provided quantitative information about cell matrix interactions in two dimensions, recent studies have shown significant differences in the behavior and morphology of cells when placed in three-dimensional environments. Hence new quantitative experimental techniques are needed to accurately determine cell traction forces in three dimensions. Recently, two approaches both based on laser scanning confocal microscopy have emerged to address this need. This study highlights the details, implementation and advantages of such a three-dimensional imaging methodology with the capability to compute cellular traction forces dynamically during cell migration and locomotion. An application of this newly developed three-dimensional traction force microscopy (3D TFM) technique to single cell migration studies of 3T3 fibroblasts is presented to show that this methodology offers a new quantitative vantage point to investigate the three-dimensional nature of cell-ECM interactions.
Highlights
The exchange of physical forces in cell-cell and cell-matrix interactions plays a significant role in regulating a variety of physiological and pathological processes including wound healing, angiogenesis, metastasis and embryogenesis [1,2,3]
This study presents a three-dimensional traction force microscopy (3D TFM) technique capable of measuring cellular deformations in three dimensions with submicron accuracy
Full field displacement measurements were carried out using the laser scanning confocal microscopy (LSCM)-digital volume correlation (DVC) technique applied to migrating 3T3 fibroblast cells on polyacrylamide gels with a Young’s modulus of E *9.64 kPa
Summary
The exchange of physical forces in cell-cell and cell-matrix interactions plays a significant role in regulating a variety of physiological and pathological processes including wound healing, angiogenesis, metastasis and embryogenesis [1,2,3]. Quantification and understanding of the nature of cell-ECM interactions and regulation within three-dimensional environments become important for the development of new biomaterials and clinical diagnostics. Within the last few decades, studies have begun to quantify traction forces that are developed by migrating cells through a variety of techniques. In 1995 Oliver et al and Dembo et al developed a quantitative technique called traction force microscopy (TFM) to study fibroblast migration on two-dimensional substrate surfaces [8,9,10]. While other experimental techniques, such as micropillars and embedded force sensors have made significant contributions in quantifying cellmatrix interactions [2,11], traction force microscopy remains the most widely used methods for measuring cellular traction forces [12,13,14,15,16]
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