High resolution traction force microscopy based on experimental and computational advances

Cell adhesion and migration crucially depend on the transmission of actomyosin-generated forces through sites of focal adhesion to the extracellular matrix. Here we report experimental and computational advances in improving the resolution and reliability of traction force microscopy. First, we intr...

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Bibliographic Details
Main Authors: Sabass, Benedikt (Author) , Schwarz, Ulrich S. (Author)
Format: Article (Journal)
Language:English
Published: 2008
In: Biophysical journal
Year: 2008, Volume: 94, Issue: 1, Pages: 207-220
ISSN:1542-0086
DOI:10.1529/biophysj.107.113670
Online Access:Verlag, kostenfrei, Volltext: http://dx.doi.org/10.1529/biophysj.107.113670
Verlag, kostenfrei, Volltext: http://www.sciencedirect.com/science/article/pii/S000634950870780X
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Author Notes:Benedikt Sabass, Margaret L. Gardel, Clare M. Waterman, and Ulrich S. Schwarz
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Summary:Cell adhesion and migration crucially depend on the transmission of actomyosin-generated forces through sites of focal adhesion to the extracellular matrix. Here we report experimental and computational advances in improving the resolution and reliability of traction force microscopy. First, we introduce the use of two differently colored nanobeads as fiducial markers in polyacrylamide gels and explain how the displacement field can be computationally extracted from the fluorescence data. Second, we present different improvements regarding standard methods for force reconstruction from the displacement field, which are the boundary element method, Fourier-transform traction cytometry, and traction reconstruction with point forces. Using extensive data simulation, we show that the spatial resolution of the boundary element method can be improved considerably by splitting the elastic field into near, intermediate, and far field. Fourier-transform traction cytometry requires considerably less computer time, but can achieve a comparable resolution only when combined with Wiener filtering or appropriate regularization schemes. Both methods tend to underestimate forces, especially at small adhesion sites. Traction reconstruction with point forces does not suffer from this limitation, but is only applicable with stationary and well-developed adhesion sites. Third, we combine these advances and for the first time reconstruct fibroblast traction with a spatial resolution of ∼1μm.
Item Description:Gesehen am 12.12.2017
Physical Description:Online Resource
ISSN:1542-0086
DOI:10.1529/biophysj.107.113670