Parallel Fourier ptychographic microscopy for high-throughput screening with 96 cameras (96 Eyes)
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.
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.Attached Files
Published - s41598-019-47146-z.pdf
Submitted - 547265.full.pdf
Supplemental Material - 41598_2019_47146_MOESM1_ESM.pdf
Supplemental Material - 41598_2019_47146_MOESM2_ESM.txt
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Additional details
- Alternative title
- 96 Eyes: Parallel Fourier Ptychographic Microscopy for high throughput screening
- PMCID
- PMC6668459
- Eprint ID
- 92931
- Resolver ID
- CaltechAUTHORS:20190214-102530450
- Caltech
- AMGEN.96EYES
- Created
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2019-02-14Created from EPrint's datestamp field
- Updated
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2023-06-01Created from EPrint's last_modified field