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Digital Inline Holographic Microscopy DIHM of Weakly-scattering Subjects
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Wide-field three-dimensional optical imaging using temporal focusing for holographically trapped microparticles.

Roman Spesyvtsev, Helen A Rendall, Kishan Dholakia

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    This study introduces a novel method for optical imaging and stimulation of cells and particles in 3D. It enables precise, depth-resolved control for applications in colloidal science and optogenetics.

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    Area of Science:

    • Optics and Photonics
    • Biophysics
    • Colloidal Science

    Background:

    • Simultaneous optical imaging or stimulation of small numbers of cells or colloidal particles in arbitrary geometries presents a significant challenge.
    • Existing methods often lack precise control over depth and spatial arrangement.

    Purpose of the Study:

    • To demonstrate a method for depth-resolved two-photon imaging and stimulation of trapped objects in arbitrary 3D geometries.
    • To enable independent position control of multiple objects using holographic beam shaping.
    • To achieve wide-field, depth-selective activation of fluorescent samples.

    Main Methods:

    • Utilizing temporal focusing with holographic optical tweezers.
    • Employing holographic beam shaping for independent position control of multiple trapped objects.
    • Demonstrating wide-field, depth-resolved illumination of trapped fluorescent beads and HL60 cells.

    Main Results:

    • Achieved depth-resolved two-photon imaging of trapped objects in arbitrary 3D geometries with a single objective.
    • Demonstrated full 3D positioning control for multiple trapped objects.
    • Showcased wide-field, depth-resolved illumination compatible with scattering media.

    Conclusions:

    • The developed approach offers precise depth control and wide-area photoactivation for colloidal science and optogenetics.
    • This technique facilitates targeted manipulation and imaging of microscale objects in complex arrangements.
    • The method is robust and adaptable for studies involving scattering media.