Single molecule detection and sequencing using fluorescence lifetime imaging
OwnerJoseph Beechem, Theofilos Kotseroglou +2inventors
InventorsJoseph Beechem, Theofilos Kotseroglou, William Michael Lafferty, Mark F. Oldham
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Abstract
A nucleic acid detection system and method are provided, in which excitation energy is transmitted from a pulsed excitation source to a reaction site including a fluorescence resonance energy transfer (FRET)-based dye system to generate a fluorescent signal at the reaction site, the fluorescent signal is detected by a detector from the reaction site, and detection of the fluorescent signal is respectively blocked and permitted at the detector by a detector gate this is timed based on an emission start time of the transmitted excitation energy.
Claims22
A nucleic acid detection system, comprising:
- a pulsed excitation source transmitting excitation energy to a reaction site including a fluorescence resonance energy transfer (FRET)-based dye system to generate a fluorescent signal at the reaction site;
- a detector configured to detect the fluorescent signal from the reaction site;
- and
- a detector gate configured to respectively block and permit detection of the fluorescent signal at the detector, said detector gate being timed based on an emission start time of the transmitted excitation energy.
A method of detecting a nucleic acid molecule sequence, comprising:
- reacting at a reaction site a nucleic acid molecule with a fluorescence resonance energy transfer (FRET)-based dye system;
- turning on an excitation source and transmitting excitation energy to the FRET-based dye system to generate fluorescent emissions at the reaction site;
- preventing detection, by a detector gate, of the fluorescent emissions at a detector after an emission start time of the transmitted excitation energy;
- turning off the excitation source;
- permitting detection of the fluorescent emissions after a first predetermined amount of time from the emission start time has elapsed;
- detecting the fluorescent emissions with the detector to form a detected signal;
- and
- determining a character or sequence of the DNA molecule based on the detected signal.
A method of detecting a nucleic acid molecule sequence, comprising:
- reacting at a reaction site a nucleic acid molecule with a fluorescence resonance energy transfer (FRET)-based dye system;
- transmitting excitation energy to the reaction site to excite the FRET-based dye system to generate fluorescent emissions at the reaction site;
- and
- syncing timing of the excitation energy transmission with detection of the fluorescent emissions by a detector and detecting the fluorescent emissions at the detector after a predetermined delay time from an emission start time of the transmitted excitation energy.
Dependencies are inferred from the claim text.