Microscopic particle localization in 3D and in multicolor (Conference Presentation)

Yoav Shechtman

doi:10.1117/12.2506391

4 March 2019 Microscopic particle localization in 3D and in multicolor (Conference Presentation)

Yoav Shechtman

Proceedings Volume 10930, Advanced Fabrication Technologies for Micro/Nano Optics and Photonics XII; 109300I (2019) https://doi.org/10.1117/12.2506391
Event: SPIE OPTO, 2019, San Francisco, California, United States

Abstract

Precise determination of the position of a single point source (e.g. fluorescent molecule/protein, quantum dot) is at the heart of microscopy methods such as single particle tracking and super-resolution localization microscopy ((F)PALM, STORM). Localizing a point source in all three dimensions, i.e. including depth, poses a significant challenge; the depth of field of a standard high-NA microscope is fundamentally limited, and its point-spread-function (PSF), namely, the shape that a point source creates in the image plane, contains little information about the emitter’s depth. Various techniques exist that enable 3D localization, prominent among them being PSF engineering, in which the PSF of a microscope is modified to encode the depth of the source. This is achieved by shaping the wavefront of the light emitted from the sample, using a phase mask in the pupil (Fourier) plane of the microscope. In this talk I will describe the use of PSF engineering for encoding the depth and color of a sub-diffraction fluorescent emitter. I will present experimental results of tracking and imaging synthetic fluorescent sources and bio-molecules, and discuss the unique fabrication considerations and challenges associated with these applications. Different methods of phase modulation will be considered and evaluated.

Conference Presentation

Citation Download Citation

Yoav Shechtman "Microscopic particle localization in 3D and in multicolor (Conference Presentation)", Proc. SPIE 10930, Advanced Fabrication Technologies for Micro/Nano Optics and Photonics XII, 109300I (4 March 2019); https://doi.org/10.1117/12.2506391

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