title: Multiplexed 3D FRET imaging in deep tissue of live embryos.
abstract: Current deep tissue microscopy techniques are mostly restricted to intensity mapping of fluorophores, which significantly limit their applications in investigating biochemical processes in vivo. We present a deep tissue multiplexed functional imaging method that probes multiple Förster resonant energy transfer (FRET) sensors in live embryos with high spatial resolution. The method simultaneously images fluorescence lifetimes in 3D with multiple excitation lasers. Through quantitative analysis of triple-channel intensity and lifetime images, we demonstrated that Ca(2+) and cAMP levels of live embryos expressing dual FRET sensors can be monitored simultaneously at microscopic resolution. The method is compatible with a broad range of FRET sensors currently available for probing various cellular biochemical functions. It opens the door to imaging complex cellular circuitries in whole live organisms.
authors.affiliation: College of Optical Sciences, University of Arizona, 1630 East University Blvd., Tucson, AZ 85721, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Department of Pediatrics and Communicable Diseases, University of Michigan, 1150 W. Medical Center Drive, Ann Arbor, MI 48109, USA.
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Wan
authors.affiliation: College of Optical Sciences, University of Arizona, 1630 East University Blvd., Tucson, AZ 85721, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Pediatrics and Communicable Diseases, University of Michigan, 1150 W. Medical Center Drive, Ann Arbor, MI 48109, USA.
authors.forename: Weibin
authors.initials: W
authors.lastname: Zhou
authors.affiliation: College of Optical Sciences, University of Arizona, 1630 East University Blvd., Tucson, AZ 85721, USA.|Department of Molecular and Cellular Biology, University of Arizona, 1007 E. Lowell St., Tucson, AZ 85721, USA.
authors.forename: Leilei
authors.initials: L
authors.lastname: Peng
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002477:Cells / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D015027:Zebrafish / Q000196:embryology*
chemical_list: D005456:Fluorescent Dyes