Snapshot volumetric imaging with engineered point-spread functions

Olesker, D., Harvey, A. R. and Taylor, J. M. (2022) Snapshot volumetric imaging with engineered point-spread functions. Optics Express, 30(19), pp. 33490-33501. (doi: 10.1364/OE.465113)

Abstract

The biological world involves intracellular and intercellular interactions that occur at high speed, at multiple scales and in three dimensions. Acquiring 3D images, however, typically requires a compromise in either spatial or temporal resolution compared to 2D imaging. Conventional 2D fluorescence imaging provides high spatial resolution but requires plane-by-plane imaging of volumes. Conversely, snapshot methods such as light-field microscopy allow video-rate imaging, but at the cost of spatial resolution. Here we introduce 3D engineered point-spread function microscopy (3D-EPM), enabling snapshot imaging of real-world 3D extended biological structures while retaining the native resolution of the microscope in space and time. Our new computational recovery strategy is the key to volumetrically reconstructing arbitrary 3D structures from the information encapsulated in 2D raw EPM images. We validate our technique on both point-like and extended samples, and demonstrate its power by imaging the intracellular motion of chloroplasts undergoing cyclosis in a sample of Egeria densa. Our technique represents a generalised computational methodology for 3D image recovery which is readily adapted to a diverse range of existing microscopy platforms and engineered point-spread functions. We therefore expect it to find broad applicability in the study of rapid biological dynamics in 3D.

• Item Type: Articles
• Additional Information: Funding: Engineering and Physical Sciences Research Council (EP/M028135/1, EP/N509668/1, EP/R513222/1).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Olesker, Mr Daniel and Taylor, Dr Jonathan and Harvey, Professor Andy
• Authors: Olesker, D., Harvey, A. R., and Taylor, J. M.
• College/School: College of Science and Engineering > School of Physics and Astronomy
• Journal Name: Optics Express
• Publisher: Optica
• ISSN: 1094-4087
• ISSN (Online): 1094-4087
• Published Online: 29 August 2022
• Copyright Holders: Copyright © 2022 The Author(s)
• First Published: First published in Optics Express 30(19): 33490-33501
• Publisher Policy: Reproduced under a Creative Commons License