Random access parallel microscopy.

Mishal Ashraf,Kiamehr Rahemipour,Simon Thibault,Byu Ri Sim,Alexander D Corbett,Denis Brousseau,Anthony Tam,Sharika Mohanan,Gil Bub

doi:10.7554/elife.56426

Mishal Ashraf, Kiamehr Rahemipour + Show 7 more

Open Access

https://doi.org/10.7554/elife.56426

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Journal: eLife	Publication Date: Jan 12, 2021
Citations: 14	License type: CC BY 4.0

Affiliation: McGill University, Université Laval, University of Exeter

Abstract

We introduce a random-access parallel (RAP) imaging modality that uses a novel design inspired by a Newtonian telescope to image multiple spatially separated samples without moving parts or robotics. This scheme enables near-simultaneous image capture of multiple petri dishes and random-access imaging with sub-millisecond switching times at the full resolution of the camera. This enables the RAP system to capture long-duration records from different samples in parallel, which is not possible using conventional automated microscopes. The system is demonstrated by continuously imaging multiple cardiac monolayer and Caenorhabditis elegans preparations.

Highlights

Conventional multi-sample imaging modalities either require movement of the sample to the focal plane of the imaging system (Klimas et al, 2016; Yemini et al, 2013; Kopljar et al, 2017; Hortigon-Vinagre et al, 2018), movement of the imaging system itself (Likitlersuang et al, 2012; Hansen et al, 2010), or use a wide-field approach to capture several samples in one frame (Larsch et al, 2013; Taute et al, 2015)
Schemes that move the sample or the imaging system can be mechanically complex and are inherently slow, while wide-field imaging systems have poor light collection efficiency and resolution compared to systems that image a single sample at a given time point
The random-access parallel (RAP) system uses a large parabolic reflector and objective lenses positioned at their focal distances above each sample

Summary

Introduction

Conventional multi-sample imaging modalities either require movement of the sample to the focal plane of the imaging system (Klimas et al, 2016; Yemini et al, 2013; Kopljar et al, 2017; Hortigon-Vinagre et al, 2018), movement of the imaging system itself (Likitlersuang et al, 2012; Hansen et al, 2010), or use a wide-field approach to capture several samples in one frame (Larsch et al, 2013; Taute et al, 2015). Schemes that move the sample or the imaging system can be mechanically complex and are inherently slow, while wide-field imaging systems have poor light collection efficiency and resolution compared to systems that image a single sample at a given time point. A fast light-emitting diode (LED) array sequentially illuminates samples to generate images that are captured with a single camera placed at the focal point of the reflector. This optical configuration allows each sample to fill a sensor’s field of view. Since each LED illuminates a single sample and LED switch times are very fast, images from spatially separated samples can be captured at rates limited only by the camera’s frame rate or the system’s ability to store data. We demonstrate the system in two low-magnification, low-resolution settings using single-element lenses and other sourced components

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