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CN-121999254-A - Automatic calibration method and system of DNA synthesis device

CN121999254ACN 121999254 ACN121999254 ACN 121999254ACN-121999254-A

Abstract

The invention discloses an automatic calibration method and an automatic calibration system for a DNA synthesis device, and belongs to the field of synthesis biological devices. Through accurate discernment and correction chip placement error and chip processing error, realize automatic calibration, get rid of the dependence to the manual experience. By optimizing the calibration flow, the single-chip calibration time is greatly shortened, and the method is suitable for mass production scenes of high-flux DNA synthesis. The micron-scale correction of chip placement errors and processing errors is realized, the alignment precision of the printed pattern and the reactive sites is ensured, and the harsh requirement of DNA synthesis is met, so that the problems of low synthesis quality, poor efficiency and the like caused by error runaway in the prior art are solved.

Inventors

  • WEN ZHIMING
  • CHEN LI
  • ZHANG PENGQI

Assignees

  • 捷莱弗科技(杭州)有限公司

Dates

Publication Date
20260508
Application Date
20260127

Claims (10)

  1. 1. An automatic calibration method of a DNA synthesis device is characterized by comprising the following steps: S1, preprocessing a DNA chip, namely marking a preset mark on a preset reference position on the surface of the DNA chip, measuring the relative relation between the center of the preset mark and a reaction site on the DNA chip, and obtaining offset error data of the reaction site; S2, calibrating the focal length and the resolution of the industrial camera, and establishing the relation among a pixel coordinate system, an image coordinate system and a camera coordinate system; the method comprises the steps of calibrating the position of an industrial camera to enable a preset identification center on a DNA chip to be located at the center of a camera visual field, establishing a DNA chip coordinate system by taking the preset identification center as an origin, and establishing the relation between the DNA chip coordinate system and a pixel coordinate system, an image coordinate system and a camera coordinate system; S3, establishing a relation between a DNA chip coordinate system and a printing coordinate system of a printing device, wherein the printing device prints a preset pattern on the DNA chip; S4, acquiring the preset marks and the preset patterns on the DNA chip by using the calibrated industrial camera; s5, acquiring an actual printing offset according to the relative position relation between the preset mark and the preset pattern; S6, adjusting printing parameters of the printing device according to the actual printing offset and the reactive site offset error data; And S7, printing on the reaction sites is completed according to the adjusted printing parameters.
  2. 2. The method of claim 1, wherein the predetermined mark is a cross mark and the predetermined pattern is an annular dot array surrounding the cross mark.
  3. 3. The method of claim 2, wherein step S1 comprises measuring the relative position between the cross marker center and a preset reaction site on the DNA chip by using a high-precision imager to obtain the offset error data of the reaction site caused by mask plate processing.
  4. 4. The method according to claim 1, wherein step S5 comprises comparing the actual position relationship between the preset mark and the preset pattern with the position relationship preset when the printing device prints, thereby obtaining the offset of the actual printing.
  5. 5. The automatic calibration method according to claim 4, wherein the step S5 comprises the steps of realizing high-precision identification by adopting a weighted template matching algorithm during image identification, and improving identification robustness in a complex environment by giving high weight to pattern edges and feature point areas and giving low weight to background interference areas.
  6. 6. The automatic calibration method according to claim 5, wherein the step S5 of realizing high-precision identification by adopting a weighted template matching algorithm further comprises the steps of carrying out region decomposition on the image according to structural features, and extracting feature subregions with high stability and rich information content; in the matching process, carrying out normalization correlation matching on each subarea respectively, and distributing weights according to the contribution degree of the subareas in the overall structure positioning; And the matching results of all the subareas are combined in a weighting manner, and the overall position and the attitude of the target are restored by utilizing geometric constraint, so that the high-precision positioning insensitive to local shielding is realized.
  7. 7. The method according to claim 1, wherein the number of the preset marks in the step S1 is two, namely a preset mark A and a preset mark B; in the step S2, two industrial cameras are provided, namely an industrial camera A and an industrial camera B, the center of a preset mark A is positioned at the center of the visual field of the industrial camera A, the center of the preset mark B is positioned at the center of the visual field of the industrial camera B, and a DNA chip coordinate system is established by taking the center of the preset mark A as an origin; In the step S3, printing two preset patterns on the DNA chip, wherein the two preset patterns are a preset pattern A and a preset pattern B respectively; In the step S4, an industrial camera A acquires a preset mark A and a preset pattern A, and an industrial camera B acquires a preset mark B and a preset pattern B; in step S5, the actual printing offset is obtained according to the relative positional relationship between the preset mark a and the preset mark B and the preset pattern a and the preset pattern B.
  8. 8. The method according to claim 7, wherein in step S2, the industrial camera A is located directly above the preset mark A, and the industrial camera B is located directly above the preset mark B, and the vertical heights of the industrial camera A and the industrial camera B are the same.
  9. 9. An automatic calibration system for a DNA synthesis apparatus, characterized in that the automatic calibration system employs the method of any one of claims 1 to 8.
  10. 10. The automatic calibration system of claim 9, wherein the DNA synthesis apparatus comprises a stage, a printing apparatus, a DNA chip, an industrial camera, and a control unit.

Description

Automatic calibration method and system of DNA synthesis device Technical Field The invention relates to the technical field of synthesis biology, in particular to an automatic calibration method and system of a DNA synthesis device. Background In the field of synthetic biology, automatic synthesis of DNA is an emerging method of DNA synthesis in the field of synthetic biology. Current DNA synthesis devices apply the principle of inkjet printing to DNA synthesis processes. Just like an ink-jet printer which sprays ink onto paper to form a pattern, the DNA synthesis system of the ink-jet technology sprays different nucleotide monomer solutions onto the surface of a specific solid-phase carrier through tiny nozzles, and sequentially adds nucleotides according to a preset sequence to gradually synthesize the required DNA fragments. As shown in fig. 5, the conventional DNA synthesizer includes a stage 100, a printer 500, a chip 200, an industrial camera, and a control unit 600. The stage 100 is used for fixing the chip 200, the stage 100 is an electric stage 100, fine adjustment can be performed, and adjustment can be performed in the horizontal direction, the longitudinal direction and the vertical direction. Printing apparatus 500 for adding nucleotides. The existing printing apparatus 500 is generally controlled by five-axis linkage, wherein the five axes include three linear axes, which may be an X axis, a Y axis, and a Z axis, and two rotation axes, which may be an R axis and an a axis, respectively. The five-axis linkage control is a common control mode, such as CN115097785A, CN102173211B, and the printing devices are all controlled by adopting a five-axis linkage mode. The DNA chip 200 is usually provided with reaction sites in an array on the chip 200. The control of the synthesis device is accomplished by a control unit. In the synthesis of the DNA synthesis apparatus, it is first necessary to prepare a suitable solid phase carrier such as a glass chip or the like and to chemically treat the surface thereof so as to fix the nucleotide. Next, according to DNA sequence information to be synthesized, a printing head of a printing apparatus is controlled to precisely spray a specific nucleotide monomer solution to a specified position on the surface of the carrier. One nucleotide at a time is sprayed and is linked to the nucleotide chain which has been linked to the carrier by a chemical reaction. The above spraying and ligating process is repeated, and different nucleotides are added in sequence until the complete DNA fragment is synthesized. After each addition of a nucleotide, washing and the like are usually also required to remove unreacted substances and impurities. The applicant's self-filed patent CN 121135805A discloses that calibration of inkjet printing is better performed by MARK and identification of reaction sites. In the inkjet printing process, due to factors such as chip placement errors, chip processing errors, manufacturing accuracy of a printing head, installation errors, working environment changes, abrasion after long-time use and the like, deviation between the actual positions of ink points and preset target positions can be caused. These problems seriously affect the quality and accuracy of inkjet printing, reducing the success rate of DNA synthesis. In the above inkjet printing process, the accuracy of the error is required to reach the micrometer level to satisfy the requirement of the reaction. The traditional calibration method generally depends on manual experience, so that the calibration process is complex, the efficiency is low, the calibration precision is difficult to ensure, and the requirement of DNA synthesis on high-precision ink-jet printing cannot be met. Therefore, there is a need for a method and system that can automatically, efficiently and accurately calibrate a DNA synthesis apparatus. Disclosure of Invention The present invention is directed to alleviating or solving at least one aspect or point of the problems discussed above. The invention provides an automatic calibration method of a DNA synthesis device, comprising the following steps of S1, preprocessing a DNA chip, namely marking a preset mark on a preset reference position on the surface of the DNA chip, measuring the relative relation between the center of the preset mark and a reaction site on the DNA chip, and obtaining offset error data of the reaction site; S2, calibrating the focal length and the resolution of the industrial camera, and establishing the relation among a pixel coordinate system, an image coordinate system and a camera coordinate system; the method comprises the steps of calibrating the position of an industrial camera to enable a preset identification center on a DNA chip to be located at the center of a camera visual field, establishing a DNA chip coordinate system by taking the preset identification center as an origin, and establishing the relation between the DNA chip coordina