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9 March 2020 Imaging 3D orientation and wobbling of single fluorescent emitters by polarized super-resolved microscopy (Conference Presentation)
Valentina Curcio, Caio Vaz Rimoli, Cesar Augusto Valades Cruz, Pascal Verdier-Pinard, Manos Mavrakis, Sophie Brasselet
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Proceedings Volume 11246, Single Molecule Spectroscopy and Superresolution Imaging XIII; 112460V (2020) https://doi.org/10.1117/12.2545668
Event: SPIE BiOS, 2020, San Francisco, California, United States
Abstract
Measuring 3D orientation properties of single fluorescent emitters including their angle wobbling, as well as their position, is a challenge that would enrich super-resolution techniques with structural molecular information. We present a polarized microscopy technique that provides all 3D orientation parameters unambiguously, using four-polarization splitting of the image plane and intensity filtering in the back focal plane. Using an inverse problem approach we can retrieve 3D orientation, wobbling and 2D position of the fluorophores with high precision. We validated the technique using fluorescent nano-beads and applied it to the structural study of fluorescently labelled F-actin filaments.
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Valentina Curcio, Caio Vaz Rimoli, Cesar Augusto Valades Cruz, Pascal Verdier-Pinard, Manos Mavrakis, and Sophie Brasselet "Imaging 3D orientation and wobbling of single fluorescent emitters by polarized super-resolved microscopy (Conference Presentation)", Proc. SPIE 11246, Single Molecule Spectroscopy and Superresolution Imaging XIII, 112460V (9 March 2020); https://doi.org/10.1117/12.2545668
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KEYWORDS
3D image processing
3D metrology
Microscopy
Polarized microscopy
Stereoscopy
Molecules
Proteins
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