Real-Time Analysis and Visualization for Single-Molecule Based Super-Resolution Microscopy |
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| Authors: | Adel Kechkar Deepak Nair Mike Heilemann Daniel Choquet Jean-Baptiste Sibarita |
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| Institution: | 1. University of Bordeaux, Interdisciplinary Institute for Neuroscience, Bordeaux, France.; 2. CNRS UMR 5297, Bordeaux, France.; 3. Goethe-University Frankfurt, Institute of Physical and Theoretical Chemistry, Frankfurt, Germany.; Humboldt University, Germany, |
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| Abstract: | Accurate multidimensional localization of isolated fluorescent emitters is a time consuming process in single-molecule based super-resolution microscopy. We demonstrate a functional method for real-time reconstruction with automatic feedback control, without compromising the localization accuracy. Compatible with high frame rates of EM-CCD cameras, it relies on a wavelet segmentation algorithm, together with a mix of CPU/GPU implementation. A combination with Gaussian fitting allows direct access to 3D localization. Automatic feedback control ensures optimal molecule density throughout the acquisition process. With this method, we significantly improve the efficiency and feasibility of localization-based super-resolution microscopy. |
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