Prof. Nathalie Westbrook Profile

Prof. Nathalie Westbrook

Professor at Institut d'Optique Graduate School/CNRS

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Publications (3)

SPIE Journal Paper | 11 July 2023 Open Access

Förster resonance energy transfer efficiency measurements on vinculin tension sensors at focal adhesions using a simple and cost-effective setup

Camille Dubois, Ludivine Houel-Renault, Marie Erard, Nada Boustany, Nathalie Westbrook

JBO, Vol. 28, Issue 08, 082808, (July 2023) https://doi.org/10.1117/12.10.1117/1.JBO.28.8.082808

KEYWORDS: Fluorescence resonance energy transfer, Focal adhesions, Calibration, Sensors, Tunable filters, Resonance energy transfer, Optical filters, Cameras, Photobleaching, Energy efficiency

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SignificanceForces inside cells play a fundamental role in tissue growth, affecting important processes such as cancer cell migration or tissue repair after injury. Förster resonance energy transfer (FRET)-based tension sensors are a remarkable tool for studying these forces and should be made easier to use.AimWe prove that absolute FRET efficiency can be measured on a simple setup, an order of magnitude more cost-effective than a standard FRET microscopy setup, by applying it to vinculin tension sensors (VinTS) at the focal adhesions of live CHO-K1 cells.ApproachOur setup located at Université Paris-Saclay acquires donor and acceptor fluorescence in parallel on two low-cost CMOS cameras and uses two LEDs for rapid switching of the excitation wavelength at a reduced cost. The calibration required to extract FRET efficiency was achieved using a single construct (TSMod). FRET efficiencies were measured for VinTS and the tail-less control VinTL, lacking the actin-binding domain of vinculin. Measurements were confirmed on the same cell type using a more standard intensity-based setup located at Rutgers University.ResultsThe average FRET efficiency of VinTS (22.0 % ± 4 % ) over more than 10,000 focal adhesions is significantly lower (p < 10 − 6) than that of VinTL (30.4 % ± 5 % ), our control that is insensitive to force, in agreement with the force exerted on vinculin at focal adhesions. Attachment of the CHO-K1 cells on fibronectin decreases FRET efficiency, thus increasing the force, compared with poly-lysine. FRET efficiency for the VinTL control is consistent with all measurements currently available in the literature, confirming the validity of our measurements and hence of our simpler setup.ConclusionsForce measurements, resolved spatially inside a cell, can be achieved using FRET-based tension sensors with a cost effective intensity-based setup. This will facilitate combining FRET with techniques for applying controlled forces such as optical tweezers.

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Proceedings Article | 9 September 2019 Presentation

Precise calibration of optical tweezers using the interference pattern in the backscattered signal and application to the microrheology of blood clots (Conference Presentation)

Nathalie Westbrook, Flavie Gillant, Julien Moreau, Karen Perronet, Maximilian Richly, Antigoni Alexandrou, Laura Wolff, Jean-Marc Allain

Proceedings Volume 11083, 110830H (2019) https://doi.org/10.1117/12.2530645

KEYWORDS: Microscopes, Bayesian inference, Spectrum analysis, Acoustics, Calibration, Precision calibration, Optical tweezers, Blood, Optical calibration, Dielectrics

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Proceedings Article | 16 September 2014 Paper

Optical tweezers calibration with Bayesian inference

Silvan Türkcan, Maximilian Richly, Antoine Le Gall, Nicolas Fiszman, Jean-Baptiste Masson, Nathalie Westbrook, Karen Perronet, Antigoni Alexandrou

Proceedings Volume 9164, 916415 (2014) https://doi.org/10.1117/12.2064375

KEYWORDS: Calibration, Bayesian inference, Optical tweezers, Diffusion, Optical calibration, Particles, Objectives, Receptors, Photodiodes, Microscopes

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