Cutrale, Francesco and Trivedi, Vikas and Trinh, Le A. and Chiu, Chi-Li and Choi, John M. and Artiga, Marcela S. and Fraser, Scott E. (2017) Hyperspectral phasor analysis enables multiplexed 5D in vivo imaging. Nature Methods, 14 (2). pp. 149-152. ISSN 1548-7091. https://resolver.caltech.edu/CaltechAUTHORS:20170117-091624181
Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20170117-091624181
Abstract
Time-lapse imaging of multiple labels is challenging for biological imaging as noise, photobleaching and phototoxicity compromise signal quality, while throughput can be limited by processing time. Here, we report software called Hyper-Spectral Phasors (HySP) for denoising and unmixing multiple spectrally overlapping fluorophores in a low signal-to-noise regime with fast analysis. We show that HySP enables unmixing of seven signals in time-lapse imaging of living zebrafish embryos.
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Additional Information: | © 2017 Macmillan Publishers Limited, part of Springer Nature. Received 18 July 2016 Accepted 08 November 2016 Published online 09 January 2017. The authors thank T.V. Truong, C. Arnesano, M. Kitano, S. Restrepo (Translational Imaging Center, University of Southern California) and G.H. Bearman for helpful discussions. This work was supported by grants from the Moore Foundation, the Coulter Foundation and the NIH (R01 HD075605, R01 OD019037). We thank C. Paquette for fish care. Author Contributions: F.C. and V.T. performed experiments and analyzed the results. J.M.C., L.A.T. and S.E.F. helped in the experimental design and data analysis. V.T. derived equations. F.C. wrote the software. M.S.A. prepared the samples. V.T., F.C., L.A.T., C.-L.C. and S.E.F. wrote the manuscript. Competing Financial Interests: The authors declare competing financial interests: details are available in the online version of the paper. | ||||||||||||
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Subject Keywords: | Developmental biology; Fluorescence imaging; Image processing | ||||||||||||
Issue or Number: | 2 | ||||||||||||
Record Number: | CaltechAUTHORS:20170117-091624181 | ||||||||||||
Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20170117-091624181 | ||||||||||||
Official Citation: | Hyperspectral phasor analysis enables multiplexed 5D in vivo imaging Francesco Cutrale, Vikas Trivedi, Le A Trinh, Chi-Li Chiu, John M Choi, Marcela S Artiga & Scott E Fraser Nature Methods 14, 149–152 (2017) doi:10.1038/nmeth.4134 | ||||||||||||
Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | ||||||||||||
ID Code: | 73517 | ||||||||||||
Collection: | CaltechAUTHORS | ||||||||||||
Deposited By: | Ruth Sustaita | ||||||||||||
Deposited On: | 17 Jan 2017 19:11 | ||||||||||||
Last Modified: | 03 Oct 2019 16:28 |
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