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Table of contents
1 General introduction
2 Force production and regulation by the cytoskeleton
2.1 The actin network
2.2 Role of molecular motors
2.3 α-actinin
2.4 Focal adhesions
2.5 Stress fibers formation and dynamics
2.6 Actomyosin cortex
3 Traction force measurement methods:
3.1 Substrate deformation based:
3.2 Cantilever based methods
3.3 Molecular sensors
4 Positioning of the study
5 Methods
5.1 Pattern design
5.2 Mask design
5.3 Silanized coverslip preparation
5.4 Passivation of fluorescent beads
5.5 Polyacrylamide micropatterning
5.6 Cell culture
5.7 Imaging
5.8 Protocol for fixed cell assay
5.9 Fluorescent labelling of cells
5.10 Image processing of fluorescent images
5.11 Translation and rotation correction functions
5.12 Traction force microscopy analysis
5.13 Automated acquisition
5.14 Transfection protocol
5.15 Photoconversion experiment protocol
5.16 Quantification of photoconverted signal in cells
5.17 Data plotting and statistical analysis
6 Results
6.1 Cell cycle analysis
6.2 Actin dynamics results
6.3 Biochemical characterization (Paper draft)
6.4 Extra data
7 General conclusions and perspectives about the work presented in this thesis manuscript.
7.1 The modified TFM assay:
7.2 Biochemical composition study
7.3 Actin dynamics results cannot explain difference in cell forces.
7.4 Perspectives
8 Abbreviations
9 Bibliography
