Flexible and uniform illumination for wide field microscopy from TIRF to epiflourescence excitation (Conference Presentation)

Adrien Mau; Nicolas Bourg; Sandrine Leveque-Fort

doi:10.1117/12.2545231

9 March 2020 Flexible and uniform illumination for wide field microscopy from TIRF to epiflourescence excitation (Conference Presentation)

Adrien Mau, Nicolas Bourg, Sandrine Leveque-Fort

Author Affiliations +

Proceedings Volume 11245, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII; 112450B (2020) https://doi.org/10.1117/12.2545231
Event: SPIE BiOS, 2020, San Francisco, California, United States

Abstract

We propose a new hybrid scanning/wide field excitation implementation, developed on a classical inverted wide field fluorescence microscope. Compatible from epifluorescence to TIRF excitation it provides flexible and uniform illumination over wide field of views (200x200 µm²), with uniform sectioning depth. This permits to enhance TIRF imaging and control photobleaching in classical microscopy. In single molecule localization microscopy, one can enhance the localization precision by balancing the density of activated fluorophores and the photon count, and extract information from many quantitative datas thanks to the wide uniform illumination. The required laser power to achieve efficient blinking is also considerably reduced.

Conference Presentation

Citation Download Citation

Adrien Mau, Nicolas Bourg, and Sandrine Leveque-Fort "Flexible and uniform illumination for wide field microscopy from TIRF to epiflourescence excitation (Conference Presentation)", Proc. SPIE 11245, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXVII, 112450B (9 March 2020); https://doi.org/10.1117/12.2545231

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