Droplet Microfluidic Flow Cytometer For Sorting On Transient Cellular Responses Of Genetically-Encoded Sensors

• Published in: Analytical chemistry (Washington) 2017-01, Vol.89 (1), p.711-719
• Main Authors: Fiedler, Brett L, Van Buskirk, Steven, Carter, Kyle P, Qin, Yan, Carpenter, Margaret C, Palmer, Amy E, Jimenez, Ralph
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Biosensing Techniques; Biosensors; Cells; Droplets; Flow Cytometry; Fluorescence; Fluorescence Resonance Energy Transfer; Fluorescent Dyes - chemistry; HeLa Cells; Humans; Measuring instruments; Microfluidic Analytical Techniques; Microfluidics; Particle Size; Populations; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Sensors; Sorting; Zinc - analysis
• ISSN: 0003-2700; 1520-6882; 1520-6882
• DOI: 10.1021/acs.analchem.6b03235

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.