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Parallel Fourier ptychographic microscopy for high-throughput screening with 96 cameras (96 Eyes)

Chan, Antony C. S. and Kim, Jinho and Pan, An and Xu, Han and Nojima, Dana and Hale, Christopher and Wang, Songli and Yang, Changhuei (2019) Parallel Fourier ptychographic microscopy for high-throughput screening with 96 cameras (96 Eyes). Scientific Reports, 9 . Art. No. 11114. ISSN 2045-2322. PMCID PMC6668459. https://resolver.caltech.edu/CaltechAUTHORS:20190214-102530450

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Abstract

We report the implementation of a parallel microscopy system (96 Eyes) that is capable of simultaneous imaging of all wells on a 96-well plate. The optical system consists of 96 microscopy units, where each unit is made out of a four element objective, made through a molded injection process, and a low cost CMOS camera chip. By illuminating the sample with angle varying light and applying Fourier Ptychography, we can improve the effective brightfield imaging numerical aperture of the objectives from 0.23 to 0.3, and extend the depth of field from ±5 μm to ±15 μm. The use of Fourier Ptychography additionally allows us to computationally correct the objectives’ aberrations out of the rendered images, and provides us with the ability to render phase images. The 96 Eyes acquires raw data at a rate of 0.7 frame per second (all wells) and the data are processed with 4 cores of graphical processing units (GPUs; GK210, Nvidia Tesla K80, USA). The system is also capable of fluorescence imaging (excitation = 465 nm, emission = 510 nm) at the native resolution of the objectives. We demonstrate the capability of this system by imaging S1P_1-eGFP-Human bone osteosarcoma epithelial (U2OS) cells.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1038/s41598-019-47146-zDOIArticle
https://doi.org/10.1101/547265DOIDiscussion Paper
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6668459PubMed CentralArticle
ORCID:
AuthorORCID
Chan, Antony C. S.0000-0002-5104-261X
Hale, Christopher0000-0002-2360-7759
Yang, Changhuei0000-0001-8791-0354
Alternate Title:96 Eyes: Parallel Fourier Ptychographic Microscopy for high throughput screening
Additional Information:© 2019 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 15 February 2019; Accepted 11 July 2019; Published 31 July 2019. We would like to thank Daniel Martin for fabricating the Siemens star phase target, and to thank Albert Chung for the suggestion of an adaptive step size for more accurate pupil recovery. We also thank Kevin Wong and Richard Graham from ClearBridge Biophotonics for the technical advices on coding the image stitching algorithm and background interference removal. Caltech Agency Award (AMGEN.96EYES). Author Contributions: Conceived and designed the hardware/software/experiments: A.C.S.C., J.K. and A.P. Contributed reagents/materials/analysis tools: H.X., D.N. and C.H. Wrote the paper: A.C.S.C. and A.P. Supervised the project: S.W. and C.Y. The authors declare no competing interests.
Funders:
Funding AgencyGrant Number
CaltechAMGEN.96EYES
PubMed Central ID:PMC6668459
Record Number:CaltechAUTHORS:20190214-102530450
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190214-102530450
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:92931
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:14 Feb 2019 18:34
Last Modified:03 Oct 2019 20:49

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