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US 8,802,424 B2Granted
Methods and systems for analysis of fluorescent reactions with modulated excitation
AssigneePacific Biosciences of California, Inc.
InventorsKhai Luong, Paul Lundquist, Ravi Dalal, John Lyle, Stephen Turner
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Abstract
Methods, systems and their components for monitoring fluorescent signals and particularly transient fluorescent signals from reaction mixtures of interest, which methods and systems employ modulated excitation light sources to reduce impacts of excessive illumination on the reaction components or the data obtained therefrom.
Claims20
A system for analysis of fluorescent materials, comprising:
- a reaction region for containing a fluorescent reaction mixture;
- at least first and second excitation light sources;
- at least one detector;
- and
- an optical train for directing excitation light from the at least first and second excitation light sources to the reaction region and collecting fluorescent signals from the reaction region and directing the fluorescent signals to the detector,
- wherein the optical train comprises a beam modulation component that modulates the excitation light from the at least first and second excitation light sources, the optical train is configured to provide at least first and second modulated beams of excitation light to the reaction region at different times, the optical beam modulation component is configured to modulate the excitation light at a frequency of between about 20 Hz and about 1000 Hz, and
- the detector is configured to detect each type of the fluorescent signals corresponding to each of the at least first and second modulated beams of the excitation light at different times, wherein the detector is synchronized with the at least first and second modulated beams of excitation light to have a sampling rate that is greater than the frequency of modulation of the at least first and second modulated beams, thus substantially reducing the background noise from reflected excitation lights and autofluorescence.
4 more elements
A system for analysis of fluorescent materials, comprising:
- a reaction region containing a reaction mixture that comprises at least first and second fluorescent reactants, the first and second fluorescent reactants having at least first and second distinct excitation spectra;
- at least first and second excitation light sources configured to provide at least first and second excitation light beams at the at least first and second excitation spectra, respectively;
- and
- an optical train for modulating the at least first and second excitation light beams from the at least first and second excitation light sources at a frequency of at least 50 Hz to provide at least first and second modulated excitation light beams and for directing the at least first and second modulated excitation light beams to the reaction region at different times,
- wherein a detector is configured to detect each type of the fluorescent signals corresponding to each of the at least first and second modulated beams of the excitation light at different times,
- wherein the detector is synchronized with the at least first and second modulated beams of excitation light to have a sampling rate that is greater than the frequency of modulation of the at least first and second modulated beams, thus substantially reducing the background noise from reflected excitation lights and autofluorescence.
3 more elements
Dependencies are inferred from the claim text.