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Holographically Informed Fluorescent Imaging (HIFI) is used to generate 3D fluorescent images. A hologram is acquired via Phase Shifted Digital Holography (PSDH), followed by a fluorescent image. The hologram is numerically propagated, generating a 3D distribution. The optimal depth of focus for localized regions are determined using localized variance in the holographic 3D space. The fluorescent image is correlated with the hologram brought into a single plane and is propagated back to the localized depth of focus, generating a 3D fluorescence image. As a proof of principle, fluorescent labeled polystyrene microspheres are used to generate 3D fluorescent images via HIFI.
Vincent M. Rossi
"Generating 3D fluorescent images via holographically informed fluorescent imaging (Conference Presentation)", Proc. SPIE PC12385, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXX, PC123850H (15 March 2023); https://doi.org/10.1117/12.2649390
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Vincent M. Rossi, "Generating 3D fluorescent images via holographically informed fluorescent imaging (Conference Presentation)," Proc. SPIE PC12385, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXX, PC123850H (15 March 2023); https://doi.org/10.1117/12.2649390