High-throughput Single-Molecule Spectroscopy in Free Solution

Abstract

A high-speed high-throughput single-molecule imaging technique for identifying molecules in free solution based on differences in their fluorescence emission spectra is presented. Unlike previous reports, the entire spectrum, rather than selected wavelengths through optical filters, is recorded. Furthermore, the millisecond data acquisition time means that the molecules do not need to be immobilized or spatially confined. In one example, individual λDNA molecules labeled with YOYO-I, POPO-III, or a combination of the two dyes can be distinguished from one another. In another example, biotinylated 2.1- kb DNA labeled with YOYO-I was reacted with avidinconjugated R-phycoerythrin. The two different reactant molecules and the product molecule can be simultaneously imaged and identified by their spectroscopic characteristics. This technique can therefore be used for screening single molecules for disease markers and for monitoring individual molecular interactions at a rate of thousands of molecules per second.

Department(s)

Chemistry

Sponsor(s)

National Institutes of Health (U.S.)

Keywords and Phrases

DNA; Fluorescent Dye; Chemical Analysis; DNA Determination; Fluorescence; Molecular Interaction; Molecule; Spectroscopy; Biotinylation; Signal Processing, Computer-Assisted; Solutions; Spectrometry, Fluorescence

International Standard Serial Number (ISSN)

0003-2700

Document Type

Article - Journal

Document Version

Citation

File Type

text

Language(s)

English

Rights

© 2000 American Chemical Society (ACS), All rights reserved.

Publication Date

01 Oct 2000

Link to Full Text

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