Depth-enhanced high-throughput microscopy by compact PSF engineering

Nadav Opatovski, Elias Nehme, Noam Zoref, Ilana Barzilai, Reut Orange Kedem, Boris Ferdman, Paul Keselman, Onit Alalouf and Yoav Shechtman ()
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Nadav Opatovski: Technion - Israel Institute of Technology
Elias Nehme: Technion - Israel Institute of Technology
Noam Zoref: Technion - Israel Institute of Technology
Ilana Barzilai: Technion - Israel Institute of Technology
Reut Orange Kedem: Technion - Israel Institute of Technology
Boris Ferdman: Technion - Israel Institute of Technology
Paul Keselman: Sartorius Stedim North America Inc.
Onit Alalouf: Technion - Israel Institute of Technology
Yoav Shechtman: Technion - Israel Institute of Technology

Nature Communications, 2024, vol. 15, issue 1, 1-11

Abstract: Abstract High-throughput microscopy is vital for screening applications, where three-dimensional (3D) cellular models play a key role. However, due to defocus susceptibility, current 3D high-throughput microscopes require axial scanning, which lowers throughput and increases photobleaching and photodamage. Point spread function (PSF) engineering is an optical method that enables various 3D imaging capabilities, yet it has not been implemented in high-throughput microscopy due to the cumbersome optical extension it typically requires. Here we demonstrate compact PSF engineering in the objective lens, which allows us to enhance the imaging depth of field and, combined with deep learning, recover 3D information using single snapshots. Beyond the applications shown here, this work showcases the usefulness of high-throughput microscopy in obtaining training data for deep learning-based algorithms, applicable to a variety of microscopy modalities.

Date: 2024
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DOI: 10.1038/s41467-024-48502-y

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