title: High-speed multiparameter photophysical analyses of fluorophore libraries.
abstract: There is a critical need for high-speed multiparameter photophysical measurements of large libraries of fluorescent probe variants for imaging and biosensor development. We present a microfluidic flow cytometer that rapidly assays 10(4)-10(5) member cell-based fluorophore libraries, simultaneously measuring fluorescence lifetime and photobleaching. Together, these photophysical characteristics determine imaging performance. We demonstrate the ability to resolve the diverse photophysical characteristics of different library types and the ability to identify rare populations.
authors.affiliation: †BioFrontiers Institute, University of Colorado, Boulder, Colorado 80309, United States.|‡Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, United States.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Dean
authors.affiliation: §Department of Physics, University of Tennessee Knoxville, Knoxville, Tennessee 37996, United States.|∥Center for Laser Applications, University of Tennessee Space Institute, Tullahoma, Tennessee 37388, United States.
authors.forename: Lloyd M
authors.initials: LM
authors.lastname: Davis
authors.affiliation: ‡Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, United States.|⊥JILA, NIST, and University of Colorado, Boulder, Colorado 80309, United States.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Lubbeck
authors.affiliation: ‡Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, United States.|⊥JILA, NIST, and University of Colorado, Boulder, Colorado 80309, United States.
authors.forename: Premashis
authors.initials: P
authors.lastname: Manna
authors.affiliation: ‡Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, United States.|⊥JILA, NIST, and University of Colorado, Boulder, Colorado 80309, United States.
authors.forename: Pia
authors.initials: P
authors.lastname: Friis
authors.affiliation: †BioFrontiers Institute, University of Colorado, Boulder, Colorado 80309, United States.|‡Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, United States.
authors.forename: Amy E
authors.initials: AE
authors.lastname: Palmer
authors.affiliation: ‡Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309, United States.|⊥JILA, NIST, and University of Colorado, Boulder, Colorado 80309, United States.
authors.forename: Ralph
authors.initials: R
authors.lastname: Jimenez
mesh_terms: D005434:Flow Cytometry / Q000295:instrumentation*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D056656:Lab-On-A-Chip Devices; D038761:Photobleaching; D013050:Spectrometry, Fluorescence
chemical_list: D005456:Fluorescent Dyes