High-speed in vitro intensity diffraction tomography

Abstract

We demonstrate a label-free, scan-free {\it intensity} diffraction tomography technique utilizing annular illumination (aIDT) to rapidly characterize large-volume 3D refractive index distributions in vitro. By optimally matching the illumination geometry to the microscope pupil, our technique reduces the data requirement by 60$\times$ to achieve high-speed 10 Hz volume rates. Using 8 intensity images, we recover $\sim 350\times 100\times 20\mu$m${}^3$ volumes with near diffraction-limited lateral resolution of 487 nm and axial resolution of 3.4 $\mu$m. Our technique's large volume rate and high resolution enables 3D quantitative phase imaging of complex living biological samples across multiple length scales. We demonstrate aIDT's capabilities on unicellular diatom microalgae, epithelial buccal cell clusters with native bacteria, and live \emph{Caenorhabditis elegans} specimens. Within these samples, we recover macro-scale cellular structures, subcellular organelles, and dynamic micro-organism tissues with minimal motion artifacts. Quantifying such features has significant utility in oncology, immunology, and cellular pathophysiology, where these morphological features are evaluated for changes in the presence of disease, parasites, and new drug treatments. Finally, we simulate our aIDT system to highlight the accuracy and sensitivity of our technique. aIDT shows promise as a powerful high-speed, label-free computational microscopy technique applications where natural imaging is required to evaluate environmental effects on a sample in real-time. We provide example datasets and an open source implementation of aIDT at \href{https://github.com/bu-cisl/IDT-using-Annular-Illumination}{https://github.com/bu-cisl/IDT-using-Annular-Illumination}.

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