Interaction between intracellular compartments is based on complex vesicular transport processes. Fluorescence based techniques now reach single molecule resolution in living cells. To avoid fluorophore photobleaching limitations, quantitative phase imaging techniques are conventionally used to produce long-term high contrast cell images.
We used an improved wave front sensing set-up called QWLSI-HD, which resolution is high enough to reach nanometric resolution for vesicle localization.
We also added specificity to quantitative phase imaging by training artificial intelligence algorithms to recognize intracellular vesicles. We compared performances between the algorithms and also between fixed and living samples.
We propose to study the behavior of vesicles at the intracellular scale with a non-invasive quantitative phase imaging technology which enables to follow vesicles during a large amount of time without modifying their behavior. We developed specific tools to localize and track cells that we compare to fluorescence-based tracking method. We studied their interaction with the cytoskeleton by inhibiting it using drugs and we implemented statistical tools to quantify and understand intracellular processes at the vesicular level. The resolution and the high sensitivity brought by the High Definition wave front sensor we developed enables to follow all individual vesicles through time follow up, while imaging the whole cell.
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