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Published August 1, 2016 | Published
Journal Article Open

Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy

Abstract

Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period.

Additional Information

© 2016 Optical Society of America. Received 23 Apr 2016; revised 7 Jul 2016; accepted 17 Jul 2016; published 22 Jul 2016. Funding: National Institute of Health (NIH) (R01AI6226-01); Caltech Agency Award (AMGEN.96EYES). We would like to thank Xiaoze Ou and Jaebum Chung for helpful discussions, and Daniel Martin for fabrication of a Siemens star phase target. Jinho Kim's work was supported by the Kwanjeong Educational Foundation.

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August 20, 2023
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