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- W3133180711 abstract "Our goal is to map chromatin dynamics in live cell nuclei by tracking 3D lattices of photoactivated chromatin microdomains. We take a multifocal imaging approach to deduce 3D microdomain motions from 2D camera images. Here we report initial results to implement a novel spiral phase plate (SPP) to achieve the 3D-to-2D mapping. The SPP is a diffractive optical element (DOE) designed to create a rotating point spread function (rPSF) in our optical microscope. The SPP consists of a series of helical ramps that provide orbital angular momentum to the emitted fluorescent light. This translates into a pattern of rPSFs in 2D (on a camera chip) that reveals, through their individual centers and angles of rotation, the 3D locations of the chromatin microdomains. We will report on the characterization of the SPP and our initial attempts at tracking chromatin microdomains in 3D. We will compare our 3D tracking results to previous measurements of 2D diffusion in the same U2OS cell line. We will also report on our use of supervised machine learning to accurately identify the source plane along the optical axis of the microscope of each of the observed fluorescent nuclear microdomains (z-position). This technique, combining a SPP and deep learning, should be applicable to any sample consisting of fluorescent objects transversely separated by several microns, such as fluorescently labeled chromatin microdomains in our project." @default.
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- W3133180711 date "2021-02-01" @default.
- W3133180711 modified "2023-09-26" @default.
- W3133180711 title "Spiral Phase Plate for Tracking 3D Motion in Cells" @default.
- W3133180711 doi "https://doi.org/10.1016/j.bpj.2020.11.1269" @default.
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