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Adam Wax

Publications and source records attributed to Adam Wax.

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Structured illumination microscopy for dual-modality 3D sub-diffraction resolution fluorescence and refractive-index reconstruction

Though structured illumination (SI) microscopy is a popular imaging technique conventionally associated with fluorescent super-resolution, recent works have suggested its applicability towards sub-diffraction coherent imaging with quantitative endogenous biological contrast. Here, we demonstrate that SI can efficiently integrate the principles of fluorescent super-resolution and coherent synthetic aperture to achieve 3D dual-modality sub-diffraction resolution, fluorescence and refractive-index (RI) visualizations of biological samples. Such a demonstration can enable future biological studies to synergistically explore molecular and biophysical/biochemical mechanisms at sub-diffraction resolutions. We experimentally demonstrate this framework by introducing a SI microscope capable of 3D sub-diffraction fluorescence and RI imaging, and verify its biological visualization capabilities by experimentally reconstructing 3D RI/fluorescence visualizations of fluorescent calibration microspheres as well as alveolar basal epithelial adenocarcinoma (A549) and human colorectal adenocarcinmoa (HT-29) cells, fluorescently stained for F-actin.

q-bio.QM

Structured illumination multimodal 3D-resolved quantitative phase and fluorescence sub-diffraction microscopy

Sub-diffraction resolution imaging has played a pivotal role in biological research by visualizing key, but previously unresolvable, sub-cellular structures. Unfortunately, applications of far-field sub-diffraction resolution are currently divided between fluorescent and coherent-diffraction regimes, and a multimodal sub-diffraction technique that bridges this gap has not yet been demonstrated. Here we report that structured illumination (SI) allows multimodal sub-diffraction imaging of both coherent quantitative-phase (QP) and fluorescence. Due to the conventional fluorescent applications of SI, we first demonstrated the principle of SI-enabled three-dimensional (3D) QP sub-diffraction imaging with calibration microspheres. Image analysis confirmed enhanced lateral and axial resolutions over diffraction-limited QP imaging, and established striking parallels between coherent SI and conventional optical diffraction tomography. We next introduce an optical system utilizing SI to achieve 3D sub-diffraction, multimodal QP/fluorescent visualization of A549 biological cells fluorescently tagged for F-actin. Our results suggest that SI has unique utility in studying biological phenomena with significant molecular, biophysical, and biochemical components.

physics.bio-ph

Refractive index tomography with structured illumination

This work introduces a novel reinterpretation of structured illumination (SI) microscopy for coherent imaging that allows three-dimensional imaging of complex refractive index (RI). To do so, we show that coherent SI is mathematically equivalent to a superposition of angled illuminations. It follows that raw acquisitions for standard SI-enhanced quantitative-phase images can be processed into complex electric-field maps describing sample diffraction under angled illuminations. Standard diffraction tomography (DT) computation can then be used to reconstruct the sample 3D RI distribution at sub-diffraction resolutions. We demonstrate this concept by using a SI-quantitative-phase imaging system to computationally reconstruct 3D RI distributions of human breast (MCF-7) and colorectal (HT-29) adenocarcinoma cells. Our experimental setup uses a spatial light modulator to generate structured patterns at the sample and collects angle-dependent sample diffraction using a common-path, off-axis interference configuration with no moving components. Furthermore, this technique holds promise for easy pairing with SI fluorescence microscopy, and important future extensions may include multimodal, sub-diffraction resolution, 3D RI and fluorescent visualizations.

physics.optics