High-throughput platform for real-time monitoring of biological processes by multicolor single-molecule fluorescence.
Proc Natl Acad Sci U S A
; 111(2): 664-9, 2014 Jan 14.
Article
in En
| MEDLINE
| ID: mdl-24379388
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.
Full text:
1
Database:
MEDLINE
Main subject:
Biophysics
/
Software
/
High-Throughput Screening Assays
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Fluorescence
/
Monitoring, Physiologic
Language:
En
Journal:
Proc Natl Acad Sci U S A
Year:
2014
Type:
Article