Hyperspectral phasor analysis enables multiplexed 5D in vivo imaging
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.
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.Attached Files
Supplemental Material - nmeth.4134-S1.pdf
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Supplemental Material - nmeth.4134-sv7.mp4
Supplemental Material - nmeth.4134-sv8.mp4
Supplemental Material - nmeth.4134-sv9.mp4
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Additional details
- Eprint ID
- 73517
- Resolver ID
- CaltechAUTHORS:20170117-091624181
- Gordon and Betty Moore Foundation
- Coulter Foundation
- NIH
- R01 HD075605
- NIH
- R01 OD019037
- Created
-
2017-01-17Created from EPrint's datestamp field
- Updated
-
2021-11-11Created from EPrint's last_modified field