title: Multiplexed labeling system for high-throughput cell sorting.
abstract: Flow cytometry and fluorescence activated cell sorting techniques were designed to realize configurable classification and separation of target cells. A number of cell phenotypes with different functionalities have recently been revealed. Before simultaneous selective capture of cells, it is desirable to label different samples with the corresponding dyes in a multiplexing manner to allow for a single analysis. However, few methods to obtain multiple fluorescent colors for various cell types have been developed. Even when restricted laser sources are employed, a small number of color codes can be expressed simultaneously. In this study, we demonstrate the ability to manifest DNA nanostructure-based multifluorescent colors formed by a complex of dyes. Highly precise self-assembly of fluorescent dye-conjugated oligonucleotides gives anisotropic DNA nanostructures, Y- and tree-shaped DNA (Y-DNA and T-DNA, respectively), which may be used as platforms for fluorescent codes. As a proof of concept, we have demonstrated seven different fluorescent codes with only two different fluorescent dyes using T-DNA. This method provides maximum efficiency for current flow cytometry. We are confident that this system will provide highly efficient multiplexed fluorescent detection for bioanalysis compared with one-to-one fluorescent correspondence for specific marker detection.
authors.affiliation: School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, South Korea.
authors.forename: Seung Won
authors.initials: SW
authors.lastname: Shin
authors.affiliation: School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, South Korea.
authors.forename: Kyung Soo
authors.initials: KS
authors.lastname: Park
authors.affiliation: School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, South Korea.
authors.forename: In Hyun
authors.initials: IH
authors.lastname: Song
authors.affiliation: School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, South Korea.
authors.forename: Woo Jung
authors.initials: WJ
authors.lastname: Shin
authors.affiliation: School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, South Korea.
authors.forename: Byung Woo
authors.initials: BW
authors.lastname: Kim
authors.affiliation: Division of Vascular Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul 06351, South Korea.
authors.forename: Dong-Ik
authors.initials: DI
authors.lastname: Kim
authors.affiliation: School of Chemical Engineering, Sungkyunkwan University, Suwon 440-746, South Korea; SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon 440-746, South Korea. Electronic address: sh.um@skku.edu.
authors.forename: Soong Ho
authors.initials: SH
authors.lastname: Um
mesh_terms: D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D003124:Colorimetry; D004269:DNA, Bacterial / Q000737:chemistry; D005434:Flow Cytometry / Q000295:instrumentation; D013194:Staining and Labeling / Q000295:instrumentation / Q000379:methods*
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA
keywords: Cell sorting; DNA nanostructure; Fluorescence-activated cell sorting; Multilabeled fluorescence marker