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In this chapter, we present the basic physical principles of the fluorescence anisotropy imaging microscopy (FAIM) and its application to study FP-tagged protein dynamics and interaction in live cells. The Förster mechanism of electronic energy transfer can occur between like chromophores (homo-fluorescence resonance energy transfer, homo-FRET) inducing(More)
Amphiphysin 1, an endocytic adaptor concentrated at synapses that couples clathrin-mediated endocytosis to dynamin-dependent fission, was also shown to have a regulatory role in actin dynamics. Here, we report that amphiphysin 1 interacts with N-WASP and stimulates N-WASP- and Arp2/3-dependent actin polymerization. Both the Src homology 3 and the N-BAR(More)
Quantitative analysis in Förster resonance energy transfer (FRET) imaging studies of protein-protein interactions within live cells is still a challenging issue. Many cellular biology applications aim at the determination of the space and time variations of the relative amount of interacting fluorescently tagged proteins occurring in cells. This relevant(More)
Fö rster Resonance Energy Transfer (FRET) measured with Fluorescence Lifetime Imaging Microscopy (FLIM) is a powerful technique to investigate spatio-temporal regulation of protein-protein interactions in living cells. When using standard fitting methods to analyze time domain FLIM, the correct estimation of the FRET parameters requires a high number of(More)
Förster Resonance Energy Transfer (FRET) measured with Fluorescence Lifetime Imaging Microscopy (FLIM) is a powerful technique to investigate spatio-temporal regulation of protein-protein interactions in living cells. When using standard fitting methods to analyze time domain FLIM, the correct estimation of the FRET parameters requires a high number of(More)
During asymmetric cell divisions, cortical dyneins generate forces essential to position the spindle after polarity cues, prescribing daughter cells fate. In nematode zygote, cortical dynein pulls on microtubules transiently, raising the question of its targeting and dynamics. Tracking and fluorescence correlation spectroscopy revealed that in the(More)
Fluorescence Lifetime Imaging Microscopy (FLIM) could investigate spatio-temporal interactions in living cells by recording the decay of the fluorescence emission at each pixel of imaged sample by phase modulation or by pulsed excitation. In the time domain time gated techniques were developed and the Phasor approach as non-fitting technic is used [1]. The(More)
Overexpression of AURKA is a major hallmark of epithelial cancers. It encodes the multifunctional serine/threonine kinase aurora A, which is activated at metaphase and is required for cell cycle progression; assessing its activation in living cells is mandatory for next-generation drug design. We describe here a Förster's resonance energy transfer (FRET)(More)
Genetically encoded Förster Resonance Energy Transfert (FRET) biosensors are powerful tools for monitoring spatiotemporal biochemical activities in living samples. By labelling a probe protein with a pair of fluorescent proteins, FRET measurement allows to follow a conformational change of the probe sensor to a specific activity. A very exciting challenge(More)
Epithelia represent a unique situation where polarized cells must maintain sufficiently strong cell-cell contacts to guarantee the epithelial integrity indispensable for barrier functions. Nevertheless, epithelia must also keep sufficient plasticity which is crucial during development and morphogenesis. Adherens junctions and mechanical forces produced by(More)