Gas-shielded trace reagent open type solid-phase synthesis gene chip synthesizer

Abstract

The invention discloses a gas protection trace reagent open type solid-phase synthesis gene chip synthesizer, which belongs to the technical field of gene chips, comprises a motor, and is characterized in that an output shaft of the motor is vertical, a vacuum adsorption turntable is fixedly arranged at the upper end of the output shaft of the motor, and a solid-phase synthesis wafer is arranged at the upper part of the vacuum adsorption turntable; a liquid adding device is arranged above the center position of the solid-phase synthesis wafer, a gas protection cover is arranged outside the solid-phase synthesis wafer, a protection gas steel cylinder is arranged outside the gas protection cover, a transition device is arranged on one side of the protection gas steel cylinder, a reagent bottle is arranged on the side, close to the protection gas steel cylinder, outside the gas protection cover, and a waste liquid bottle is arranged below the gas protection cover. According to the gene chip synthesis device disclosed by the invention, the synthesis quality of a gene chip in an open synthesis environment is effectively ensured, the dosage of a reagent is remarkably reduced, and a good structural basis is provided for integration of synthesis and photoetching and automation of a solid-phase synthesis gene chip process.

Description

Technical Field

[0001] The invention relates to the technical field of gene chips, in particular to a gas-protected trace reagent open solid-phase synthesis gene chip synthesizer. Background Art

[0002] Gene chip is a miniaturized bioanalysis platform. Its technical core lies in fixing a large number of oligonucleotide probes on the surface of a miniature solid structure to form a high-density probe array. Oligonucleotide probes are short-chain nucleic acid molecules used to detect specific DNA or RNA sequences, usually composed of 20-30 nucleotides. Gene chips can be prepared by solid-phase synthesis. Previously, they were mainly synthesized in situ using photosensitive precursors on the surface of a structure called a solid-phase wafer in an open synthesis environment. However, photosensitive precursors are extremely sensitive to oxygen and moisture in the air, which is particularly easy to affect the synthesis quality. In order to solve this problem, it was improved to use a sealed reaction chamber immersion method for synthesis. However, this method requires a large amount of synthesis reagents, and the reaction chamber is sealed, making synthesis and photolithography independent of each other. The front-end and back-end processes can only be connected manually, and the automation of the solid-phase synthesis gene chip process cannot be realized. Based on this situation, how to solve the problem that oxygen and moisture in the air of the open synthesis environment interfere with the synthesis quality and the sealed reaction chamber immersion method requires a large amount of reagents, realize the automation of the solid-phase synthesis gene chip process, and ensure the synthesis quality of the gene chip, is becoming an increasingly urgent problem for relevant technical personnel. Summary of the invention

[0003] In order to solve the above technical problems, the present invention provides a gas-protected trace reagent open solid-phase synthesis gene chip synthesizer.

[0004] A gas-protected trace reagent open solid-phase synthesis gene chip synthesizer comprises a motor, and is characterized in that: the output shaft of the motor is vertical, a vacuum adsorption turntable is fixedly arranged on the upper end of the output shaft of the motor, a solid-phase synthesis wafer is arranged on the upper part of the vacuum adsorption turntable, a liquid charger is arranged above the center position of the solid-phase synthesis wafer, a gas protection cover is arranged outside the solid-phase synthesis wafer, a protection gas cylinder is arranged outside the gas protection cover, a transition device is arranged on one side of the protection gas cylinder, a reagent bottle is arranged on the side of the gas protection cover close to the protection gas cylinder, a waste liquid bottle is arranged below the gas protection cover, a computing control board is arranged on the upper part of the motor, and the motor is electrically connected to the computing control board.

[0005] Furthermore, the output shaft of the motor is hollow, the output shaft of the motor is connected to an external vacuum source, and the solid phase synthesis wafer and the vacuum adsorption turntable are attracted by negative pressure.

[0006] Furthermore, the thickness of the reagent on the surface of the solid phase synthesis wafer ranges from 100 nm to 1000 μm.

[0007] Furthermore, a concentric liquid adding nozzle is fixedly arranged inside the liquid adder, and the lower part of the concentric liquid adding nozzle is retracted inward. The number of the concentric liquid adding nozzles is 4 groups, and the number of the concentric liquid adding nozzles in each group is no more than 4. The concentric liquid adding nozzles are arranged in a circular array, and the concentric liquid adding nozzles correspond to the reagent bottles one by one.

[0008] Furthermore, a pipeline is set between the transition device and the reagent bottle, and a driving gas pressure regulating valve is fixedly set on the pipeline. A pipeline is set between the reagent bottle and the liquid adder, and a pulse metering valve is fixedly set on the pipeline. The driving gas pressure regulating valve and the pulse metering valve are both electrically connected to the computing control board.

[0009] Furthermore, a liquid-adding visual inspection camera is fixedly arranged on one side above the solid phase synthesis wafer, and the liquid-adding visual inspection camera is inclined toward the solid phase synthesis wafer. A liquid-adding visual lighting is fixedly arranged on the other side above the solid phase synthesis wafer, and the liquid-adding visual inspection camera and the liquid-adding visual lighting are both electrically connected to the computing control board.

[0010] Furthermore, a pipeline is arranged between the transition device and the gas protection hood, and a protective gas pressure regulating valve, a protective gas heat exchanger, and a protective gas control valve are fixedly arranged in sequence on the pipeline from the transition device to the gas protection hood, and the protective gas pressure regulating valve, the protective gas heat exchanger, and the protective gas control valve are all electrically connected to the computing control board.

[0011] Furthermore, a protective gas detection sensor is arranged inside the gas protection cover, and the protective gas detection sensor is electrically connected to the computing control board. The protective gas detection sensor includes a protective gas high-level detection sensor and a protective gas low-level detection sensor. Helium is arranged inside the protective gas cylinder. The upper part of the gas protection cover is closed and the lower part is open. The protective gas low-level detection sensor is located below the solid phase synthesis wafer. The protective gas high-level detection sensor is located below the protective gas low-level detection sensor and within a reasonable buffer zone of the lower part of the gas protection cover.

[0012] Furthermore, a blow-drying nozzle is arranged on the outside of the liquid charger, and the lower part of the blow-drying nozzle is inclined toward the direction of the solid phase synthesis wafer. A pipeline is arranged between the transition device and the blow-drying nozzle. A blow-drying gas pressure regulating valve and a blow-drying nozzle switch are fixedly arranged on the pipeline from the transition device to the blow-drying nozzle in sequence, and the blow-drying gas pressure regulating valve and the blow-drying nozzle switch are both electrically connected to the computing control board.

[0013] The beneficial effects of the gas-protected trace reagent open solid-phase synthesis gene chip synthesizer of the present invention are:

[0014] 1. Simply start the motor to drive the solid phase synthesis wafer to rotate. The centrifugal force generated by the rotation process combined with the annular array of concentric liquid adding nozzles can well achieve uniform diffusion of reagents on the surface of the solid phase synthesis wafer. The reagent thickness range is controlled at 100nm-1000μm. Compared with the sealed reaction chamber immersion method, the reagent thickness is about 5mm, and the reagent dosage is significantly reduced, effectively reducing the solid phase synthesis cost of gene chips.

[0015] 2. The protective gas cylinder, transition device, pipeline, gas protection cover and helium or nitrogen form a gas protection system. The gas protection cover forms an open synthesis environment filled with inert gas and protected by inert gas, which effectively solves the problem of oxygen and moisture in the air of the open synthesis environment interfering with the synthesis quality. Combined with the adjustment of the helium or nitrogen temperature by the heat exchanger, the synthesis quality of the gene chip in the open synthesis environment is effectively guaranteed;

[0016] 3. The open synthesis environment, excellent synthesis quality, and the drying effect of the drying nozzle provide a good structural foundation for the integration of synthesis and photolithography and the automation of solid-phase synthesis gene chip process;

[0017] 4. The preset reagent supply can be obtained by controlling the driving gas pressure regulating valve and the pulse metering valve. The reagent thickness can be adjusted by adjusting the motor speed. The end position of the reagent diffusion can be controlled by fine-tuning the motor speed according to the image signal of the liquid adding visual detection camera, so that the synthesis quality of the edge gene chip can be guaranteed. According to the feedback of the protective gas detection sensor and the switch of the protective gas control valve, the solid phase synthesis wafer can always be protected by inert gas, thereby ensuring both the convenience of operation and the synthesis quality of the gene chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art, but this does not limit the protection scope of the present invention.

[0019] Figure 1 This is a schematic diagram of the present invention using helium as a protective gas;

[0020] Figure 2 This is a schematic diagram of the present invention using nitrogen as a protective gas;

[0021] Figure 3 A top view of the concentric liquid adding nozzle of the present invention;

[0022] Figure 4 It is a simplified diagram of the working scene of the liquid adding visual detection camera of the present invention.

[0023] Among them, 1-liquid adding visual inspection camera, 2-gas protection cover, 3-blow drying nozzle, 4-vacuum adsorption turntable, 5-liquid adder, 6-motor, 7-solid phase synthesis wafer, 8-protective gas low level detection sensor, 9-protective gas control valve, 10-protective gas heat exchanger, 11-reagent bottle, 12-protective gas cylinder, 13-protective gas pressure regulating valve, 14-blow drying gas pressure regulating valve, 15-drive gas pressure regulating valve, 16-pulse metering valve, 17-blow drying nozzle switch, 18-protective gas high level detection sensor, 19-transitioner, 20-waste liquid bottle, 21-concentric liquid adding nozzle, 22-liquid adding visual lighting. DETAILED DESCRIPTION

[0024] To make the description clearer, a gas-protected trace reagent open solid-phase synthesis gene chip synthesizer of the present invention is further described in conjunction with the accompanying drawings.

[0025] Example 1

[0026] A gas-protected trace reagent open solid-phase synthesis gene chip synthesizer comprises a motor 6, characterized in that: the output shaft of the motor 6 is vertical, a vacuum adsorption turntable 4 is fixedly arranged on the upper end of the output shaft of the motor 6, a solid-phase synthesis wafer 7 is arranged on the upper part of the vacuum adsorption turntable 4, a liquid charger 5 is arranged above the center position of the solid-phase synthesis wafer 7, a gas protection cover 2 is arranged outside the solid-phase synthesis wafer 7, a protection gas cylinder 12 is arranged outside the gas protection cover 2, a transition device 19 is arranged on one side of the protection gas cylinder 12, a reagent bottle 11 is arranged on the outside of the gas protection cover 2 close to the protection gas cylinder 12, a waste liquid bottle 20 is arranged below the gas protection cover 2, a computing control board is arranged on the upper part of the motor 6, and the motor 6 is electrically connected to the computing control board.

[0027] Furthermore, the output shaft of the motor 6 is hollow, and the output shaft of the motor 6 is connected to an external vacuum source, and the solid phase synthesis wafer 7 and the vacuum adsorption turntable 4 are attracted by negative pressure.

[0028] Furthermore, the thickness of the reagent on the surface of the solid phase synthesis wafer 7 ranges from 100 nm to 1000 μm.

[0029] Furthermore, a concentric liquid adding nozzle 21 is fixedly arranged inside the liquid adding device 5, and the lower part of the concentric liquid adding nozzle 21 is retracted inward. The number of the concentric liquid adding nozzles 21 is 4 groups, and the number of the concentric liquid adding nozzles 21 in each group is no more than 4. The concentric liquid adding nozzles 21 are arranged in a circular array, and the concentric liquid adding nozzles 21 correspond one-to-one to the reagent bottles 11.

[0030] Furthermore, a pipeline is set between the transition device 19 and the reagent bottle 11, and a driving gas pressure regulating valve 15 is fixedly set on the pipeline. A pipeline is set between the reagent bottle 11 and the liquid adder 5, and a pulse metering valve 16 is fixedly set on the pipeline. The driving gas pressure regulating valve 15 and the pulse metering valve 16 are both electrically connected to the computing control board.

[0031] Furthermore, a liquid-adding visual inspection camera 1 is fixedly arranged on one side above the solid phase synthesis wafer 7, and the liquid-adding visual inspection camera 1 is inclined toward the solid phase synthesis wafer 7. A liquid-adding visual lighting 22 is fixedly arranged on the other side above the solid phase synthesis wafer 7, and the liquid-adding visual inspection camera 1 and the liquid-adding visual lighting 22 are both electrically connected to the computing control board.

[0032] Furthermore, a pipeline is arranged between the transition device 19 and the gas protection hood 2, and a protective gas pressure regulating valve 13, a protective gas heat exchanger 10, and a protective gas control valve 9 are fixedly arranged in sequence on the pipeline from the transition device 19 to the gas protection hood 2, and the protective gas pressure regulating valve 13, the protective gas heat exchanger 10, and the protective gas control valve 9 are all electrically connected to the computing control board.

[0033] Furthermore, a protective gas detection sensor is arranged inside the gas protection cover 2, and the protective gas detection sensor is electrically connected to the computing control board. The protective gas detection sensor includes a protective gas high-level detection sensor 18 and a protective gas low-level detection sensor 8. Helium is arranged inside the protective gas cylinder 12. The gas protection cover 2 is closed at the upper part and opened at the lower part. The protective gas low-level detection sensor 8 is located below the solid phase synthesis wafer 7. The protective gas high-level detection sensor 18 is located below the protective gas low-level detection sensor 8 and within a reasonable buffer zone of the lower part of the gas protection cover 2.

[0034] Furthermore, a blow-drying nozzle 3 is arranged outside the liquid charger 5, and the lower part of the blow-drying nozzle 3 is inclined toward the direction of the solid phase synthesis wafer 7. A pipeline is arranged between the transition device 19 and the blow-drying nozzle 3. A blow-drying gas pressure regulating valve 14 and a blow-drying nozzle switch 17 are fixedly arranged in sequence on the pipeline from the transition device 19 to the blow-drying nozzle 3. The blow-drying gas pressure regulating valve 14 and the blow-drying nozzle switch 17 are electrically connected to the computing control board.

[0035] Example 2

[0036] In Embodiment 1, “helium is arranged inside the protective gas cylinder 12” and related descriptions, there is another design, which is described as follows.

[0037] Furthermore, nitrogen is arranged inside the protective gas cylinder 12, the upper part of the gas protective cover 2 is open and the lower part is closed, the protective gas low-level detection sensor 8 is located at a position higher than the liquid charger 5, and the protective gas high-level detection sensor 18 is located above the protective gas low-level detection sensor 8 and within a reasonable buffer zone of the upper part of the gas protective cover 2.

[0038] The working principle of the gas-protected trace reagent open solid phase synthesis gene chip synthesizer of the present invention is:

[0039] (1) Taking Example 1 as an example, the reagent supply amount is preset by the calculation control board according to the reagent amount required for synthesis and the reagent thickness on the surface of the solid phase synthesis wafer 7;

[0040] (2) The computing control board opens the protective gas pressure regulating valve 13, the protective gas heat exchanger 10, and the protective gas control valve 9, and the helium in the protective gas cylinder 12 is discharged, heated to a preset temperature by the protective gas heat exchanger 10, and then introduced into the gas protection cover 2 until the helium fills the protective gas high-level detection sensor 18 and the space above. When the helium is reduced to the position of the protective gas low-level detection sensor 8 over time, the protective gas control valve 9 automatically opens and replenishes gas into the gas protection cover 2, so that the solid phase synthesis wafer 7 and the structure above it are always protected by helium and the quality of gene chip synthesis will not be affected by oxygen and moisture in the air;

[0041] (3) The calculation control panel opens the driving gas pressure regulating valve 15 and the pulse metering valve 16, and the helium in the protective gas cylinder 12 is introduced into the reagent bottle 11. The internal gas pressure of the reagent bottle 11 increases, and the reagent is pressed to the pulse metering valve 16. When the pulse metering valve 16 is opened, the reagent flows at a set flow rate. By controlling the number of pulses of the pulse metering valve 16, the preset supply amount of reagent can be guided to the concentric liquid adding nozzle 21 and sprayed on the center position of the solid phase synthesis wafer 7;

[0042] (4) The computing control board starts the motor 6, and the output shaft of the motor 6 drives the vacuum adsorption turntable 4 and the solid phase synthesis wafer 7 to rotate as a whole. The rotating solid phase synthesis wafer 7 generates centrifugal force, and the reagent diffuses from the center position of the solid phase synthesis wafer 7 to the edge position. The conventional rotation of the solid phase synthesis wafer 7 cannot make the reagent diffuse stably to the predetermined position every time, but may end at any position. The liquid adding visual detection camera 1 feeds back the diffusion of the reagent on the surface of the solid phase synthesis wafer 7 to the computing control board. The computing control board fine-tunes the speed of the motor 6 according to the received feedback signal. When the predetermined position is not reached, the speed is increased until the reagent diffuses stably to the predetermined position. When the predetermined position has been reached, the speed is reduced and maintained for a certain time, so that the synthesis quality of the gene chip at the edge position of the solid phase synthesis wafer 7 can also be guaranteed, and a synthesis work is completed at the same time;

[0043] (5) The reagent has high transparency. If the liquid adding visual detection camera 1 is placed upright, the recognition signal will be insufficient. The inclined liquid adding visual detection camera 1 cooperates with the liquid adding visual lighting 22, combined with the difference in refractive index between the reagent position and the reagent-free position of the solid phase synthesis wafer 7, so that the signal is easy to identify, thereby completing accurate feedback on the reagent diffusion situation;

[0044] (6) The computing control board opens the blow-drying gas pressure regulating valve 14 and the blow-drying nozzle switch 17. The helium in the protective gas cylinder 12 is directed to the blow-drying nozzle 3 and blown obliquely toward the solid phase synthesis wafer 7 until the surface of the solid phase synthesis wafer 7 is dried and ready for the next photolithography process.

[0045] The above are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that are not conceived through creative work should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope defined in the claims.

Claims

1. A gas-protected trace reagent open solid-phase synthesis gene chip synthesizer, including a motor, characterized in that: The output shaft of the motor is vertical, and a vacuum adsorption turntable is fixedly arranged on the upper end of the output shaft of the motor, a solid phase synthesis wafer is arranged on the upper part of the vacuum adsorption turntable, a liquid charger is arranged above the center position of the solid phase synthesis wafer, a gas protection cover is arranged outside the solid phase synthesis wafer, a protective gas cylinder is arranged outside the gas protection cover, a transition device is arranged on one side of the protective gas cylinder, a reagent bottle is arranged on the side of the gas protection cover close to the protective gas cylinder, a waste liquid bottle is arranged below the gas protection cover, a computing control board is arranged on the upper part of the motor, and the motor is electrically connected to the computing control board.

2. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: The output shaft of the motor is hollow, and the output shaft of the motor is connected to an external vacuum source. The solid phase synthesis wafer is attracted to the vacuum adsorption turntable by negative pressure.

3. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: The thickness of the reagent on the surface of the solid phase synthesis wafer ranges from 100 nm to 1000 μm.

4. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: A concentric liquid adding nozzle is fixedly arranged inside the liquid adder, and the lower part of the concentric liquid adding nozzle is retracted inward. There are 4 groups of the concentric liquid adding nozzles, and the number of the concentric liquid adding nozzles in each group is no more than 4. The concentric liquid adding nozzles are arranged in a circular array, and the concentric liquid adding nozzles correspond to the reagent bottles one by one.

5. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: A pipeline is set between the transition device and the reagent bottle, and a driving gas pressure regulating valve is fixedly set on the pipeline. A pipeline is set between the reagent bottle and the liquid adder, and a pulse metering valve is fixedly set on the pipeline. The driving gas pressure regulating valve and the pulse metering valve are both electrically connected to the computing control board.

6. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: A liquid adding visual inspection camera is fixedly arranged on one side above the solid phase synthesis wafer, and the liquid adding visual inspection camera is tilted toward the solid phase synthesis wafer. A liquid adding visual lighting is fixedly arranged on the other side above the solid phase synthesis wafer, and the liquid adding visual inspection camera and the liquid adding visual lighting are both electrically connected to the computing control board.

7. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: A pipeline is arranged between the transition device and the gas protection hood, and a protective gas pressure regulating valve, a protective gas heat exchanger, and a protective gas control valve are fixedly arranged in sequence on the pipeline from the transition device to the gas protection hood. The protective gas pressure regulating valve, the protective gas heat exchanger, and the protective gas control valve are all electrically connected to the computing control board.

8. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: A protective gas detection sensor is arranged inside the gas protection cover, and the protective gas detection sensor is electrically connected to the computing control board. The protective gas detection sensor includes a protective gas high-level detection sensor and a protective gas low-level detection sensor. Helium is arranged inside the protective gas cylinder. The upper part of the gas protection cover is closed and the lower part is open. The protective gas low-level detection sensor is located below the solid phase synthesis wafer. The protective gas high-level detection sensor is located below the protective gas low-level detection sensor and within a reasonable buffer zone of the lower part of the gas protection cover.

9. A gas-protected trace reagent open solid phase synthesis gene chip synthesizer according to claim 1, characterized in that: A blow-drying nozzle is arranged outside the liquid charger, and the lower part of the blow-drying nozzle is inclined toward the direction of the solid phase synthesis wafer. A pipeline is arranged between the transition device and the blow-drying nozzle. A blow-drying gas pressure regulating valve and a blow-drying nozzle switch are fixedly arranged in sequence on the pipeline from the transition device to the blow-drying nozzle. The blow-drying gas pressure regulating valve and the blow-drying nozzle switch are both electrically connected to the computing control board.

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