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Computational Methods for High Resolution 3D Fluorescence Microscopy

D. Sage, F. Soulez

The BioImage Analysis Community Conference "Workflows and Open Tools" (NEUBIAS2020'17), Lisbon, Portuguese Republic, February 12-17, 2017, in press.

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Epifluorescence microscopy results in blurry images with a very coarse optical sectioning; this limits its usefulness for cellular imaging, where resolving subcellular structures requires resolution close to or even beyond Abbe's diffraction limit. Techniques such as confocal microscopy (CLSM) and selective plane illumination microscopy (SPIM) have been proposed to reduce the out-of-focus light and to improve the resolution. Several other modalities, such as single-molecule localization microscopy (SMLM) and structured illumination microscopy (SIM) make the use of multiple acquisitions, trading time for resolution. These last techniques have been recently extended to 3D imaging.

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