Abstract: The present disclosure discloses a total internal reflection fluorescence imaging system and a sequencing device. The total internal reflection fluorescence imaging system includes a first imaging system and a second imaging system. The first imaging system includes N separate laser light paths. The second imaging system determines the changes of the height of the sample by means of the differences of the positions at which the second receiving device acquires the second images, and performs automatic compensation according to the changes of the height of the sample, so as to ensure that the sample is always at a focused position. The embodiment can perform light path adjustment of the N separate laser light paths respectively by means of the second imaging system, so as to realize that the N separate laser light paths have the same penetration depth. Therefore no further light path adjustment is required when switching between different light sources, thus saving the time consuming for light path adjustment.

Abstract: A nucleic acid sequencing system is provided, including a base provided with a clamping platform, a reagent storage unit, a fluid control unit, a mobile platform and a total reflection microscope thereon; the clamping platform being provided with a gene sequencing chip thereon; the reagent storage unit being configured to store a gene sequencing reagent, the fluid control unit being configured to pump the gene sequencing reagent from the reagent storage unit to the gene sequencing chip, the mobile platform being configured to drive the clamping platform to move toward or away from the total reflection microscope; and the total reflection microscope being configured to detect a gene sequence of a sample in the gene sequencing chip.

Abstract: The present disclosure discloses a total internal reflection fluorescence imaging system and a sequencing device. The total internal reflection fluorescence imaging system includes a first imaging system and a second imaging system. The first imaging system includes N separate laser light paths. The second imaging system determines the changes of the height of the sample by means of the differences of the positions at which the second receiving device acquires the second images, and performs automatic compensation according to the changes of the height of the sample, so as to ensure that the sample is always at a focused position. The embodiment can perform light path adjustment of the N separate laser light paths respectively by means of the second imaging system, so as to realize that the N separate laser light paths have the same penetration depth. Therefore no further light path adjustment is required when switching between different light sources, thus saving the time consuming for light path adjustment.

Abstract: An embodiment of the present disclosure discloses a method and a device of correcting a single-molecule image, the method includes: acquiring the first target simplified matrix corresponding to the first image according to the intensity matrix corresponding to each of pixels in the first image, and acquiring the second target simplified matrix corresponding to the second image according to the intensity matrix corresponding to each of the pixels in the second image; the two-dimensional DFT is performed to the first target simplified matrix and the second target simplified matrix respectively to acquire the first Fourier matrix corresponding to the first target simplified matrix and the second Fourier matrix corresponding to the second target simplified matrix; the method of correcting a single-molecule image acquire the offset of the second image with respect to the first image according to the first Fourier matrix and the second Fourier matrix; and correcting the second image according to the offset.

Abstract: An embodiment of the present disclosure discloses a method and a device of correcting a single-molecule image, the method includes: acquiring the first target simplified matrix corresponding to the first image according to the intensity matrix corresponding to each of pixels in the first image, and acquiring the second target simplified matrix corresponding to the second image according to the intensity matrix corresponding to each of the pixels in the second image; the two-dimensional DFT is performed to the first target simplified matrix and the second target simplified matrix respectively to acquire the first Fourier matrix corresponding to the first target simplified matrix and the second Fourier matrix corresponding to the second target simplified matrix; the method of correcting a single-molecule image acquire the offset of the second image with respect to the first image according to the first Fourier matrix and the second Fourier matrix; and correcting the second image according to the offset.

Abstract: A sequencing method comprising: (i) combining a template nucleic acid having a first optical detection label at the end with a primer to obtain a first complex; (ii) imaging the first complex to obtain a first image; (iii) mixing the first complex, polymerase, and one or more of nucleotides with a optical detection label to obtain an extension product by polymerization reaction; (iv) imaging the extension product to obtain a second image; (v) removing the cleavable group of the nucleotides with the optical detection label from the extension product to obtain a second complex; (vi) repeating the above steps (ii) to (v) once or more times to determine the template nucleic acid sequence. The sequencing method can achieve single molecule sequencing by capturing template nucleic acids through primers.

Abstract: A nucleic acid sequencing system is provided, including a base provided with a clamping platform, a reagent storage unit, a fluid control unit, a mobile platform and a total reflection microscope thereon; the clamping platform being provided with a gene sequencing chip thereon; the reagent storage unit being configured to store a gene sequencing reagent, the fluid control unit being configured to pump the gene sequencing reagent from the reagent storage unit to the gene sequencing chip, the mobile platform being configured to drive the clamping platform to move toward or away from the total reflection microscope; and the total reflection microscope being configured to detect a gene sequence of a sample in the gene sequencing chip.