title: 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.
authors.affiliation: Translational Imaging Center, University of Southern California, Los Angeles, California, USA.|Molecular and Computational Biology, University of Southern California, Los Angeles, California, USA.
authors.forename: Francesco
authors.initials: F
authors.lastname: Cutrale
authors.affiliation: Translational Imaging Center, University of Southern California, Los Angeles, California, USA.|Molecular and Computational Biology, University of Southern California, Los Angeles, California, USA.|Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California, USA.|Department of Genetics, University of Cambridge, Cambridge, UK.
authors.forename: Vikas
authors.identifier: 0000-0003-0953-0553
authors.initials: V
authors.lastname: Trivedi
authors.affiliation: Translational Imaging Center, University of Southern California, Los Angeles, California, USA.|Molecular and Computational Biology, University of Southern California, Los Angeles, California, USA.
authors.forename: Le A
authors.initials: LA
authors.lastname: Trinh
authors.affiliation: Lawrence J. Ellison Institute for Transformative Medicine, Center for Applied Molecular Medicine, University of Southern California, Los Angeles, California, USA.
authors.forename: Chi-Li
authors.initials: CL
authors.lastname: Chiu
authors.affiliation: Translational Imaging Center, University of Southern California, Los Angeles, California, USA.|Molecular and Computational Biology, University of Southern California, Los Angeles, California, USA.
authors.forename: John M
authors.initials: JM
authors.lastname: Choi
authors.affiliation: Translational Imaging Center, University of Southern California, Los Angeles, California, USA.|Molecular and Computational Biology, University of Southern California, Los Angeles, California, USA.
authors.forename: Marcela S
authors.initials: MS
authors.lastname: Artiga
authors.affiliation: Translational Imaging Center, University of Southern California, Los Angeles, California, USA.|Molecular and Computational Biology, University of Southern California, Los Angeles, California, USA.
authors.forename: Scott E
authors.initials: SE
authors.lastname: Fraser
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D003116:Color; D004625:Embryo, Nonmammalian; D005583:Fourier Analysis; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D012984:Software; D059008:Time-Lapse Imaging / Q000379:methods*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics
chemical_list: D049452:Green Fluorescent Proteins