title: High-throughput fluorescence-activated nanoscale subcellular sorter with single-molecule sensitivity.
abstract: Recent single-cell and single-molecule studies have shown that a variety of subpopulations exist within biological systems, such as synaptic vesicles, that have previously been overlooked in common bulk studies. By isolating and enriching these various subpopulations, detailed analysis with a variety of analytical techniques can be done to further understand the role that various subpopulations play in cellular dynamics and how alterations to these subpopulations affect the overall function of the biological system. Previous sorters lack the sensitivity, sorting speed, and efficiency to isolate synaptic vesicles and other nanoscale systems. This paper describes the development of a fluorescence-activated nanoscale subcellular sorter that can sort nearly 10 million objects per hour with single-molecule sensitivity. Utilizing a near-nanoscale channel system, we were able to achieve upward of 91% recovery of desired objects with a 99.7% purity.
authors.affiliation: Department of Chemistry, University of Washington, Seattle, Washington 98195, United States.
authors.forename: Perry G
authors.initials: PG
authors.lastname: Schiro
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Gadd
authors.forename: Gloria S
authors.initials: GS
authors.lastname: Yen
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Chiu
mesh_terms: D000818:Animals; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006834:Hydrazines / Q000737:chemistry; D046210:Microfluidic Analytical Techniques; D018613:Microscopy, Confocal; D036103:Nanotechnology; D051381:Rats; D013572:Synaptic Vesicles / Q000737:chemistry* / Q000378:metabolism
chemical_list: C417561:Alexa 488 hydrazide; D004129:Dimethylpolysiloxanes; D005456:Fluorescent Dyes; D006834:Hydrazines; C013830:baysilon