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Multiplexed labeling system for high-throughput cell sorting

Author:
Shin, Seung Won, Park, Kyung Soo, Song, In Hyun, Shin, Woo Jung, Kim, Byung Woo, Kim, Dong-Ik, Um, Soong Ho
Source:
Analytical biochemistry 2016 v.508 pp. 124-128
ISSN:
0003-2697
Subject:
color, flow cytometry, fluorescence, fluorescent dyes, nanomaterials, oligonucleotides, phenotype, transfer DNA
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.
Agid:
5285861