Format

Send to

Choose Destination

See 1 citation found by title matching your search:

Rev Sci Instrum. 2014 Nov;85(11):113101. doi: 10.1063/1.4900727.

Fluorescence lifetime plate reader: resolution and precision meet high-throughput.

Author information

1
Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA.
2
Fluorescence Innovations, Inc., Minneapolis, Minnesota 55455, USA.

Abstract

We describe a nanosecond time-resolved fluorescence spectrometer that acquires fluorescence decay waveforms from each well of a 384-well microplate in 3 min with signal-to-noise exceeding 400 using direct waveform recording. The instrument combines high-energy pulsed laser sources (5-10 kHz repetition rate) with a photomultiplier and high-speed digitizer (1 GHz) to record a fluorescence decay waveform after each pulse. Waveforms acquired from rhodamine or 5-((2-aminoethyl)amino) naphthalene-1-sulfonic acid dyes in a 384-well plate gave lifetime measurements 5- to 25-fold more precise than the simultaneous intensity measurements. Lifetimes as short as 0.04 ns were acquired by interleaving with an effective sample rate of 5 GHz. Lifetime measurements resolved mixtures of single-exponential dyes with better than 1% accuracy. The fluorescence lifetime plate reader enables multiple-well fluorescence lifetime measurements with an acquisition time of 0.5 s per well, suitable for high-throughput fluorescence lifetime screening applications.

PMID:
25430092
PMCID:
PMC4242087
DOI:
10.1063/1.4900727
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for American Institute of Physics Icon for PubMed Central
Loading ...
Support Center