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Structured illumination microscopy and adaptive optics for 3D dynamic fluorescent imaging
Pierre Vermeulen, doctorant - Ph.D student Crédits : ESPCI ParisTechRecent developments in fluorescence microscopy (spatial resolution improvement, sampleinduced aberrations correction...) offer many opportunities in biological imaging. But spreading of these techniques will require reduction of the restrictions on sample preparation, fluorescent probes, utilisation complexity, or imaging frame rate.
During my Ph.D, I worked on the development and improvement of some of these techniques. Two structured illumination systems have first been built in order to accelerate optical sectioning realisation in thick samples. A second part of my work focused on the implementation of a new reconstruction approach in structured illumination microscopy to overcome the resolution limit imposed by the diffraction. This tool allows the reduction of the number
of images required for reconstruction of a super-resolution image, leading to an increase in imaging speed.
Additionally, an adaptive optics set-up has been developed to correct the sample-induced aberrations, improving imaging abilities in thick samples. The main purpose of this technique
is to provide efficient correction, while protecting the sample through the use of independent fluorescent beads for correction estimation. Finally, an instrument for fluorescence quantum
yield measurement dedicated to the characterization of infrared fluorophores has been studied to overcome the limitations of conventional methods in the infrared spectral range.