High-throughput Platform for Real-time Monitoring of Biological Processes by Multicolor Single-molecule Fluorescence

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2014 Jan 1

PMID 24379388

Citations 72

Authors

Jin Chen

Ravindra V Dalal

Alexey N Petrov

Albert Tsai

Sean E OLeary

Karen Chapin

Janice Cheng

Mark Ewan

Pei-Lin Hsiung

Paul Lundquist

Stephen W Turner

David R Hsu

Joseph D Puglisi

Affiliations

Soon will be listed here.

Abstract

Zero-mode waveguides provide a powerful technology for studying single-molecule real-time dynamics of biological systems at physiological ligand concentrations. We customized a commercial zero-mode waveguide-based DNA sequencer for use as a versatile instrument for single-molecule fluorescence detection and showed that the system provides long fluorophore lifetimes with good signal to noise and low spectral cross-talk. We then used a ribosomal translation assay to show real-time fluidic delivery during data acquisition, showing it is possible to follow the conformation and composition of thousands of single biomolecules simultaneously through four spectral channels. This instrument allows high-throughput multiplexed dynamics of single-molecule biological processes over long timescales. The instrumentation presented here has broad applications to single-molecule studies of biological systems and is easily accessible to the biophysical community.

Citing Articles

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