title: Droplet Microfluidic Flow Cytometer For Sorting On Transient Cellular Responses Of Genetically-Encoded Sensors.
abstract: Fluorescent biosensors are important measurement tools for in vivo quantification of pH, concentrations of metal ions and other analytes, and physical parameters such as membrane potential. Both the development of these sensors and their implementation in examining cellular heterogeneity requires technology for measuring and sorting cells based on the fluorescence levels before and after chemical or physical perturbations. We developed a droplet microfluidic platform for the screening and separation of cell populations on the basis of the in vivo response of expressed fluorescence-based biosensors after addition of an exogenous analyte. We demonstrate the capability to resolve the responses of two genetically encoded Zn2+sensors at a range of time points spanning several seconds and subsequently sort a mixed-cell population of varying ratios with high accuracy.
authors.affiliation: JILA, University of Colorado and NIST, Boulder Colorado 80309, United States.|Department of Chemistry and Biochemistry, University of Colorado , Boulder Colorado 80309, United States.
authors.forename: Brett L
authors.identifier: 0000-0002-1729-0185
authors.initials: BL
authors.lastname: Fiedler
authors.affiliation: JILA, University of Colorado and NIST, Boulder Colorado 80309, United States.
authors.forename: Steven
authors.initials: S
authors.lastname: Van Buskirk
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado , Boulder Colorado 80309, United States.|BioFrontiers Institute, University of Colorado , Boulder Colorado 80303, United States.
authors.forename: Kyle P
authors.initials: KP
authors.lastname: Carter
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado , Boulder Colorado 80309, United States.|Department of Biological Sciences, University of Denver , Denver, Colorado 80208, United States.
authors.forename: Yan
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado , Boulder Colorado 80309, United States.|BioFrontiers Institute, University of Colorado , Boulder Colorado 80303, United States.
authors.forename: Margaret C
authors.initials: MC
authors.lastname: Carpenter
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado , Boulder Colorado 80309, United States.|BioFrontiers Institute, University of Colorado , Boulder Colorado 80303, United States.
authors.forename: Amy E
authors.initials: AE
authors.lastname: Palmer
authors.affiliation: JILA, University of Colorado and NIST, Boulder Colorado 80309, United States.|Department of Chemistry and Biochemistry, University of Colorado , Boulder Colorado 80309, United States.
authors.forename: Ralph
authors.identifier: 0000-0002-8989-405X
authors.initials: R
authors.lastname: Jimenez
mesh_terms: D015374:Biosensing Techniques; D005434:Flow Cytometry; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D046210:Microfluidic Analytical Techniques; D010316:Particle Size; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics; D015032:Zinc / Q000032:analysis*
chemical_list: D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; C000628846:ZapCV2; C000628847:ZapCV5; D015032:Zinc