Nucleic acid sequencing by selective excitation of microparticles
First Claim
1. A method for sequencing nucleic acid microparticles, the method comprising the steps of:
- selectively exciting a calibration target by interference of a plurality of illumination beams;
obtaining optical images of the excited calibration target;
processing the optical images of the excited calibration target to generate a first optical image of the excited calibration target;
predicting a second optical image of the excited calibration target based on first calibration parameters;
generating second calibration parameters based on a difference between the first optical image and the second optical image;
performing a sequencing reaction on the nucleic acid microparticles using one or more sequencing reagents, the nucleic acid particles being different from the calibration target;
selectively exciting the nucleic acid microparticles in an excitation pattern using the second calibration parameters;
obtaining optical images of the nucleic acid microparticles at a resolution insufficient to resolve individual microparticles; and
processing the optical images of the nucleic acid microparticles using information on the excitation pattern to determine a presence or absence of at least one optical signature, the presence or absence of said at least one optical signature indicating sequence information of the nucleic acid microparticles.
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Accused Products
Abstract
Nucleic acid microparticles are sequenced by performing a sequencing reaction on the microparticles using one or more reagents, selectively exciting the microparticles in an excitation pattern, optically imaging the microparticles at a resolution insufficient to resolve individual microparticles, and processing the optical images of the microparticles using information on the excitation pattern to determine the presence or absence of the optical signature, which indicates the sequence information of the nucleic acid. An apparatus for optical excitation of the microparticles comprises an optical fiber delivering a first laser beam, and an interference pattern generation module coupled to the optical fiber. The interference pattern generation module splits the first laser beam into second and third laser beams and generates the excitation pattern for selectively exciting the microparticles by interference between the second and third laser beams.
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Citations
16 Claims
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1. A method for sequencing nucleic acid microparticles, the method comprising the steps of:
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selectively exciting a calibration target by interference of a plurality of illumination beams; obtaining optical images of the excited calibration target; processing the optical images of the excited calibration target to generate a first optical image of the excited calibration target; predicting a second optical image of the excited calibration target based on first calibration parameters; generating second calibration parameters based on a difference between the first optical image and the second optical image; performing a sequencing reaction on the nucleic acid microparticles using one or more sequencing reagents, the nucleic acid particles being different from the calibration target; selectively exciting the nucleic acid microparticles in an excitation pattern using the second calibration parameters; obtaining optical images of the nucleic acid microparticles at a resolution insufficient to resolve individual microparticles; and processing the optical images of the nucleic acid microparticles using information on the excitation pattern to determine a presence or absence of at least one optical signature, the presence or absence of said at least one optical signature indicating sequence information of the nucleic acid microparticles. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method for sequencing nucleic acid microparticles, the method comprising the steps of:
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performing a sequencing reaction on the nucleic acid microparticles using one or more sequencing reagents; selectively exciting the nucleic acid microparticles with excitation patterns generated by interference of a plurality of illumination beams, at least one of the illumination beams being modulated in only two or three phases with respect to a period of an interference pattern to generate different sinusoidal interference patterns for use as the excitation patterns; obtaining optical images of the nucleic acid microparticles at a resolution insufficient to resolve individual microparticles; and processing the optical images of the nucleic acid microparticles using information on the excitation patterns to determine a presence or absence of at least one optical signature, the presence or absence of said at least one optical signature indicating sequence information of the nucleic acid microparticles. - View Dependent Claims (15, 16)
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Specification