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Field dipole interaction and polarization effects in light-sheet optical fluorescence microscopy

Kumar, P and Mondal, PP (2023) Field dipole interaction and polarization effects in light-sheet optical fluorescence microscopy. In: Journal of Applied Physics, 133 (22).

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Official URL: https://doi.org/10.1063/5.0153038


Polarization plays a crucial role in understanding the interaction of fluorescent molecules in a light field. We report the study on the effect of a field-dipole interaction under polarization light-sheet fluorescence microscopy using the vectorial theory of light. The molecule is suitably modeled as a radiating electric dipole in a polarized electric field (both linear and random), and the system point spread function (PSF) is determined for different orientations of the dipole (both fixed and random). PSF analysis and contour plots suggest distinct nature of a field distribution in each case, indicating the importance of a field-dipole interaction for high-quality fluorescence imaging. The analysis suggests that the field spreads gradually along the polarization axis at a high numerical aperture (NA) of the objective lens, whereas it is more isotropic and homogeneous at low NA. Moreover, fast changes are not observed at low NA (i.e., far from the central lobe in the field contour plots), suggesting the absence of high-frequency components. However, sidelobes are prominent for linear polarized (along x ) light. On the other hand, rapid variations are evident for randomly polarized light, depicting the presence of high spatial frequencies in the system optical transfer function. The other significant observation is the distinct frequency spectrum (both k x and k y ) for random and fixed dipoles, indicating the significance of dipole orientation in a light-sheet field. Compared to the point-illumination-based fluorescence microscopy, sheet based polarization technique provides a high signal-to-noise ratio, a uniform field, an order large field of view, and critical information (related to the micro-environment of a dipole and its short-range interactions). The study is expected to facilitate polarization-sensitive investigation of large biological specimens (both fixed and live).

Item Type: Journal Article
Publication: Journal of Applied Physics
Publisher: American Institute of Physics Inc.
Additional Information: The copyright for this article belongs to the American Institute of Physics Inc.
Keywords: Electric fields; Fluorescence imaging; Fluorescence microscopy; Light polarization; Molecules; Quality control; Signal to noise ratio, Contour plot; Dipole interaction; Dipole polarization; Fluorescent molecules; Interaction effect; Light sheet; Numerical aperture; Optical fluorescence; Point-Spread function; Polarization effect, Optical transfer function
Department/Centre: Division of Physical & Mathematical Sciences > Instrumentation Appiled Physics
Date Deposited: 18 Jul 2023 10:23
Last Modified: 18 Jul 2023 10:23
URI: https://eprints.iisc.ac.in/id/eprint/82458

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