CN102703314A - Control system for DNA (Deoxyribose Nucleic Acid) sequencer - Google Patents

Abstract

The invention discloses a control system for a DNA (Deoxyribose Nucleic Acid) sequencer, which comprises a PLC (Programmable Logic Controller), controllers of first and second servo motors, first and second peristaltic pumps and driving motors of first and second multipass reversing valves, wherein the controllers of the first and second servo motors are electrically connected with the PLC; the first and second peristaltic pumps are respectively and electrically connected with the PLC and are used for receiving control signals of the PLC; a CCD (Charge Coupled Device) camera is electrically connected with the PLC and is used for receiving a control signal of the PLC; and a plurality of sensors are respectively and electrically connected with the PLC and are used for sending position signals of the CCD camera to the PLC. According to the control system disclosed by the invention, a reagent supply assembly of the DNA sequencer with a plurality of reaction bins timely and accurately supplies reagents and buffer solution for a plurality of reaction bins and the CCD camera can be ensured to timely read an optical signal in each reaction bin, and therefore, the aim of simultaneously performing a plurality of reactions is fulfilled, so that a plurality of samples can be simultaneously sequenced and the DNA sequencing efficiency is greatly improved.

Description

Control system for DNA sequencer

technical field

The invention relates to the technical field of DNA sequencing, in particular to a control system for a DNA sequencer.

Background technique

In the field of DNA sequencing technology, pyrosequencing technology (pyrosequencing) is a new type of enzymatic cascade sequencing technology developed by Nyren et al. in 1987. Its repeatability and accuracy performance are comparable to Sanger method DNA sequencing technology. , while the speed is greatly improved.

Pyrosequencing technology is an enzyme cascade chemiluminescent reaction in the same reaction system catalyzed by four enzymes. The principle of pyrosequencing technology is: after the primers anneal to the template DNA, the four enzymes DNA polymerase (DNA polymerase), ATP sulfurylase (ATP sulfurylase), luciferase (luciferase) and adenosine triphosphate (Apyrase) Under the synergistic effect of the primer, the polymerization of each dNTP on the primer is coupled with the release of an optical signal, and the purpose of real-time determination of DNA sequence is achieved by detecting the release and intensity of light. The reaction system of pyrosequencing technology consists of reaction substrates, single strands to be tested, sequencing primers and four enzymes. The reaction substrates are 5'-phosphosulfate (adenosine-5'-phosphosulfate, APS) and luciferin.

In each round of sequencing reaction, only one deoxynucleotide triphosphate (dNTP) is added to the reaction system. If it can just pair with the next base of the DNA template, it will be added under the action of DNA polymerase. To the 3' end of the sequencing primer, a molecule of pyrophosphate (PPi) is released at the same time. Under the action of ATP sulfurylase, the generated PPi can combine with APS to form ATP, and under the catalysis of luciferase, the generated ATP can combine with luciferin to form oxyluciferin, and at the same time generate visible light. A specific detection peak can be obtained through a weak light detection device and processing software, and the height of the peak is proportional to the number of matching bases. If the added dNTP cannot pair with the next base of the DNA template, the above reaction will not occur, and no peak will be detected. The remaining dNTPs and a small amount of residual ATP in the reaction system were degraded under the action of Apyrase. After the previous round of reaction is completed, another dNTP is added to repeat the above reaction, and the accurate DNA sequence information can be read according to the obtained peak diagram.

The overall operation process is described as follows: After the DNA sample is crushed, the library construction reagent is used to add adapters, single-strand capture, bind to microspheres, microemulsion PCR amplification, and emulsion breaking to obtain a DNA library built on microspheres. The template lays the library and the enzymes required for the sequencing reaction on the sequencing chip with a micro-reaction pool, installs the sequencing chip and sequencing reagents on the host computer, and starts the sequencing program by controlling the computer according to the number and position of the modules, and automatically performs the sequencing reaction. The data is transmitted to the data analysis computer, and after the sequencing is completed, the calculation and analysis software is used for image processing, sequence readout, quality analysis, sequence splicing, etc., and finally the sequence information of the DNA sample is obtained. The sequencing chip engraved with micro-reaction cells is obtained by etching a single-sided optical fiber core layer on an optical fiber panel with a core diameter of 25 μm and a thickness of 2 mm. The etching depth is 40 μm. There are a total of about 3 million micro-reaction cells on the chip, including about 1.2 million imaging parts micro reaction pool. The micro-reaction pool sequencing chip is the carrier of the sequencing reaction. The DNA Beads carrying the sequencing template and various enzymes for sequencing reactions are located in the sequencing chip with the micro-reaction pool engraved on it.

During the sequencing process, a chemical reaction occurs on the sequencing chip to generate visible light, and the light signal generated by the sequencing reaction is captured by a CCD (Charge Couple Device) camera to obtain the required sequencing information.

The inventors found that the DNA sequencer in the prior art has only one reaction chamber, and one instrument can only perform one test, and the work efficiency is not high. In order to improve work efficiency, the inventor has produced a DNA sequencer with multiple reaction chambers. Due to the existence of multiple reaction chambers, there are timely supply of reaction reagents and buffers for multiple reaction chambers and the use of a CCD camera to timely collect multiple Coordination of optical signals in reaction chambers.

Contents of the invention

The technical problem to be solved by the present invention is to provide a control system for a DNA sequencer with multiple reaction chambers, to control the reagent supply component to provide each reaction chamber with reaction reagents and buffers for sequencing reactions in good time, and to control the CCD camera Movement to read the light signal in each reaction chamber in time.

In order to solve the above problems, the present invention adopts the following technical solutions:

A control system for a DNA sequencer, the DNA sequencer includes a plurality of reaction chambers; a CCD camera for collecting optical signals generated by DNA sequencing reactions in each of the reaction chambers; used to support the A two-dimensionally adjustable support device for the CCD camera, the support device includes a first servo motor that drives the CCD camera to switch between different reaction chambers and a second servo motor that drives the CCD camera close to or away from one of the reaction chambers , the support device is provided with a plurality of sensors for detecting the position of the CCD camera along the direction approaching or away from the reaction chamber; and a reagent supply assembly for providing reaction reagents and buffers for a plurality of the reaction chambers , the reagent supply assembly includes a first peristaltic pump for extracting sequencing reaction reagents, a second peristaltic pump for extracting buffer, a first multi-way reversing valve for selecting reagents, and a second channel for selecting reaction chambers. Two multi-way directional valves, the first multi-way directional valve and the second multi-way directional valve are rotary valves driven by motors;

The control system includes a PLC; the PLC is electrically connected to the controller of the first servo motor and the controller of the second servo motor to control the start, stop and rotation of the first servo motor and the second servo motor direction; the PLC is electrically connected to the first peristaltic pump and the second peristaltic pump to control the start, stop and speed of the first peristaltic pump and the second peristaltic pump; the PLC is respectively connected to the first multi-channel The driving motor of the reversing valve is electrically connected with the driving motor of the second multi-way reversing valve to control the connection position of the first multi-way reversing valve and the second multi-way reversing valve; the PLC is electrically connected to the CCD camera connected to control the on and off state of the CCD camera; the PLC is also connected to the plurality of sensors to receive the position signal of the CCD camera sent by the sensor.

Preferably, the control system further includes a temperature control module, the temperature control module includes a relay connected to the PLC and a temperature sensor and a semiconductor heater arranged in the reaction chamber, and the PLC receives information from the temperature The signal of the sensor is also connected to the primary circuit of the relay, and the secondary circuit of the relay is connected to the power supply circuit of the semiconductor heater.

Preferably, the control system further includes a host computer connected to the PLC, and the host computer sets a control program run by the PLC and transmits the control program to the PLC.

Preferably, the host computer communicates with the PLC through an RS232 port.

Preferably, the host computer communicates with the CCD camera through a USB port for receiving image data acquired by the CCD camera.

The control system for a DNA sequencer of the present invention enables the reagent supply components of the DNA sequencer to supply reagents and buffers for multiple reaction chambers in a timely and accurate manner, and enables the CCD camera to read the light in each reaction chamber in a timely manner. Signals enable multiple reactions to be carried out at the same time, so that multiple samples can be sequenced at the same time, greatly improving the efficiency of DNA sequencing.

Description of drawings

FIG. 1 is a schematic diagram of a three-dimensional structure of a DNA sequencer using a control system according to an embodiment of the present invention;

2 is a schematic diagram of a reaction chamber assembly of a DNA sequencer using a control system according to an embodiment of the present invention;

Fig. 3 is the F-F direction view of Fig. 2;

4 is an exploded schematic view of one of the reaction chambers in the reaction chamber assembly shown in FIG. 2;

Fig. 5 is a schematic front view of the base of the reaction chamber shown in Fig. 4;

Fig. 6 is a left side view (partial section) schematic diagram of Fig. 5;

Fig. 7 is a structural schematic diagram of the cooperation between the reaction tank body and the guide rod of the reaction chamber shown in Fig. 4;

Fig. 8 is a schematic diagram of the left side view (partial section) of Fig. 7;

Fig. 9 is a schematic structural view of the reaction cell body in Fig. 8;

Fig. 10 is a schematic structural view of the guide rod in Fig. 8;

Fig. 11 is the A-A direction sectional view of Fig. 7;

Fig. 12 is a schematic front view of the mounting seat of the reaction chamber shown in Fig. 4;

Fig. 13 is a schematic left side view (partial section) of Fig. 12;

Fig. 14 is a schematic diagram of the B-B section of Fig. 12;

Figure 15 is a schematic diagram of the C-C direction of Figure 14;

Fig. 16 is the front view of the sliding tongue in Fig. 14;

Fig. 17 is the left side view of Fig. 16;

Fig. 18 is a schematic diagram of the three-dimensional structure of the baffle in Fig. 14;

Figure 19 is a side view of the baffle shown in Figure 18;

Figure 20 is a D-D cross-sectional view of Figure 19;

Fig. 21 is a schematic structural diagram of the CCD camera of the DNA sequencer using the control system of the embodiment of the present invention (the front end of the camera body is connected with a connecting flange);

Figure 22 is a front view of the connecting flange in Figure 21;

Figure 23 is a sectional view taken along E-E of Figure 22;

Fig. 24 is a structural schematic diagram (partial cross-section) when one of the reaction compartments of the DNA sequencer using the control system of the embodiment of the present invention is connected to a CCD camera;

Fig. 25 is a three-dimensional schematic diagram of the connection between the two-dimensionally adjustable support device and the CCD camera of the DNA sequencer using the control system of the embodiment of the present invention;

Fig. 26 is a schematic perspective view of the second linear motion mechanism of the two-dimensionally adjustable support device in Fig. 25;

Fig. 27 is a top view of the second linear motion mechanism shown in Fig. 26;

Figure 28 is a cross-sectional view along the G-G direction of Figure 27;

Figure 29 is a sectional view along the H-H direction of Figure 28;

FIG. 30 is a schematic diagram of the liquid circuit structure of the reagent supply component of the DNA sequencer of the control system of the embodiment of the present invention:

Fig. 31 is a flowchart of the control method of the reagent supply assembly shown in Fig. 30:

FIG. 32 is a schematic structural diagram of a control system for a DNA sequencer according to an embodiment of the present invention:

FIG. 33 is a schematic structural diagram of a temperature control module of a control system for a DNA sequencer according to an embodiment of the present invention (taking a reaction chamber as an example).

Explanation of main reference signs

101-support platform 103-shock absorber 102-shock absorber

104-Reaction chamber components 110-CCD camera 105-Two-dimensionally adjustable support device

1-Camera body 2-Connecting flange 4-Fiber optic panel

5-Reaction cell body 6-Mounting seat 7-Sequencing chip

8-base 9-guide rod 21-convex ring

41-Screw 44-Through Hole 45-Set

49-The third step 51-Sequencing reaction pool 54-Backboard

55-Boss 56-Annular groove 57-Heating chamber

58-Sealing ring 61-Screw 62-Sliding tongue plate

63-seat body 64-baffle 66-ring groove

67-Location ring 69-Installation cavity 80-Cavity

81-first spring 82-second through hole 84-support frame

85-spring rod 86-spring seat 87-rotating shaft

91-Connecting part 92-Sliding part 93-Protruding part

31-The first linear motion mechanism 32-The second linear motion mechanism

35-Support seat 38-Sensor

271-Second lead screw 272-Nut 281-Limiting seat

311-the first guide rail seat 312-the first guide rail 313-the first slider

319-the first limit block 321-the second guide rail seat 322-the second guide rail

323-Second slider 326-Second servo motor 327-Coupling

328-limit rod 380-groove 381-shading piece

510-Liquid inlet 511-Liquid outlet 561-Temperature sensor

571-semiconductor heater 572-closed plate 605-sliding section

606-Installation section 610-First step 612-Screw hole

621-Protrusion 623-Second Step 633-Second Spring

700-main pipeline 701-the first peristaltic pump 702-the first multi-way reversing valve

711-The second peristaltic pump 712-The second multi-way reversing valve 731-The first vacuum defoamer

732-Second vacuum defoamer 741-Reagent bottle 742-Waste liquid bucket

743 buffer bottle 750 reaction chamber 760-Tee connector

841-Screw

Detailed ways

The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments, but not as a limitation of the present invention.

Firstly, the detailed structure of the DNA sequencer adopting the control system of the embodiment of the present invention will be described in detail with reference to the accompanying drawings 1-29. The control system of this embodiment can be applied to a DNA sequencer with multiple reaction chambers, and is not limited to the DNA sequencer described in this embodiment.

As shown in Fig. 1, Fig. 2 and Fig. 3, the DNA sequencer used in the control system of the present invention includes a support platform 101, a plurality of shock absorbers 103 and a shock absorber connected to the support platform 101 through a plurality of shock absorbers 103. Shock plate 102; a reaction chamber assembly 104 for performing DNA sequencing reactions, the reaction chamber assembly 104 includes a support frame 84 vertically arranged on the shock absorbing plate and a plurality of reaction chambers arranged side by side on the support frame 84; the present embodiment , taking four reaction chambers as an example for illustration. The CCD camera 110 used to collect the optical signal generated by the DNA sequencing reaction in the reaction chamber; used to support the CCD camera 110 and drive the CCD camera 110 to approach or stay away from each reaction chamber when it is aligned with one of the reaction chambers, and when it is aligned with each reaction chamber A two-dimensionally adjustable support device 105 that can be switched between the aligned positions of each reaction chamber; a control system 109 for providing reaction reagents and buffers for the reaction chamber assembly 104; the reaction chamber assembly 104 and the two-dimensionally adjustable support device 105 are all fixedly arranged on the shock absorbing plate 102, and the control system 109 is arranged on the supporting platform 101. In this embodiment, the support platform 101 is placed on the chassis (not shown in the figure), and the instruments and circuit components used for sequencing analysis are arranged in the chassis, and the shock absorber 103 adopts an air damping shock absorber to isolate external vibrations. To eliminate or weaken its impact on the working quality of the DNA sequencer, in this embodiment, a shock absorber 103 is arranged at each of the four corners of the lower end of the shock-absorbing bottom 102 .

Firstly, the detailed structure and working process of the reaction chamber assembly 104 of the DNA sequencer applied in the control system of this embodiment will be described with reference to FIGS. 1-24 .

As shown in Fig. 2, Fig. 3, Fig. 4 and Fig. 24, in this embodiment, each reaction chamber includes a base 8 that is rotatably connected to a support frame 84, a reaction pool body 5 for performing a sequencing reaction, and a chamber for installing a sequencing reaction. The mounting seat 6 of the sequencing chip of the chip 7; the mounting seat 6 of the sequencing chip is fixedly connected on the reaction cell body 5, and the base 8 and the reaction cell body 5 are provided with a plurality of guide rods 9 passing through both of them, and the guide rods 9 is fixedly connected with the base 8, and the reaction pool body 5 is sleeved on the guide rod 9, slides in the axial direction of the guide rod 9 and can move slightly in the radial direction of the guide rod 9. A first elastic component is provided between the reaction cell body 5 and the base 8, as shown in Figure 1, in this embodiment, the first elastic component is preferably a first spring 81, the spring has the advantages of simple structure and easy installation, and of course It can be realized by selecting other elastic parts that can realize the same function.

As shown in Fig. 4, Fig. 5 and Fig. 6, one side of the base 8 is provided with a cavity 80, and two spring rods 85 are arranged side by side in the cavity 80, and the spring rod 85 is integrally located in the cavity 80, and cannot extend out of the cavity. Cavity 80. A first spring 81 is sleeved on the upper part of each spring rod 85 , and the first spring 81 extends out of the cavity 80 . Preferably, in this embodiment, a spring seat 86 is provided at the bottom of the cavity 80 , and the spring rod 85 is located on the spring seat 86 . The additional spring seat 86 can reduce the required length of the first spring 81 . The cavity 80 is provided to facilitate the installation of other components.

As shown in Fig. 4, Fig. 7 and Fig. 11, the reaction pool body 5 comprises a back plate 54 and a boss 55 which is arranged on the back plate 54 and protrudes to one side of the back plate 54, and the end surface of the boss 55 A closed annular groove 56 is arranged on the top, and the area surrounded by the closed annular groove 56 is a sequencing reaction pool 51; as shown in Figure 7, a sealing ring 58 is embedded in the annular groove 56, and the sealing ring 58 protrudes The end surface of the boss 55 is to enclose the sequencing reaction pool 51, wherein the end surface of the boss 55 is as flat as possible. As shown in Fig. 4, Fig. 7, Fig. 8 and Fig. 9, one side of the reaction cell body 5 provided with a back plate 54 is attached to the base 8, and the boss 55 of the reaction cell body 5 is provided with four first through holes , the first through hole is stepped, the section close to the base is the sliding section 605 , the other section of the first through hole is the installation section 606 , the inner diameter of the installation section 606 is larger than the sliding section 605 , forming a first step 610 . As shown in Figures 7-10, one end of the guide rod 9 is a connecting portion 91 connected to the base 8, and the other end of the guide rod 9 is provided with a raised portion 93 for being clamped at the first step 610. The guide rod 9 The part of the middle part close to the raised part 93 is the sliding part 92 for cooperating with the sliding section 605 . As shown in FIG. 12 , threads are provided on the connecting portion 91 , and as shown in FIG. 7 , the end of the protruding portion 93 of the guide rod 9 is provided with a knife groove for matching a screwdriver. As shown in Figure 1, the guide rod 9 passes through the first through hole on the reaction cell body 5 and the second through hole 82 on the base 8, so that the connecting part 91 exposes the back of the base 8, and then is fastened by a nut, thereby realizing The fixed connection between the guide rod 9 and the base 8. In order to realize sliding connection between the reaction cell body 5 and the guide rod 9, the diameter of the sliding section 605 is slightly larger than the diameter of the sliding part 92, so that the reaction cell body 5 can slide in the axial direction of the guide rod 9, and can The slight movement of the reaction cell body 5 in the radial direction of the guide rod 9 is realized. In this embodiment, as a preferred solution, the guide rod 9 is provided with a raised portion 93, so that when the reaction cell body 5 is slidably connected to the guide rod 9, the position of the raised portion 93 and the first step 610 can be limited. Function, when tightening the nut threaded with the connecting portion 91, the raised portion 93 pushes the reaction cell body 5 to move toward the base 8 to compress the first spring 81 so that there is a certain gap between the reaction cell body 5 and the base 8. The above-mentioned first spring 81 extends out of the cavity 80 so that when the reaction cell body 5 is attached to the base 8, the first spring 81 can give the reaction cell body 5 a certain force.

As shown in Fig. 4, Fig. 12, Fig. 13 and Fig. 14, the mounting seat 6 of the sequencing chip includes a seat body 63 and an installation cavity 69 opened on the seat body 63, and the mounting seat 6 of the sequencing chip is located on the reaction pool body 5 and is provided with a convex cavity. On one side of the platform 55, the seat body 63 and the back plate 54 are fixedly connected so that the boss 55 extends into the installation cavity 69, the sequencing chip 7 is fixed on the outer end of the installation cavity 69, and the end surface of the sequencing chip 7 and the boss 55 There is a gap for the flow of reagents for sequencing, which is sealed by a sealing ring 58 embedded in the annular groove 56. The size of the gap is about 0.2mm, and the sealing ring 58 protrudes from the annular groove 56 and the sequencing Chips 7 are in contact to form a closed sequencing reaction pool 51 for sequencing reaction.

As shown in Fig. 7 and Fig. 11, in this embodiment, the annular groove 56 is hexagonal, and the two ends of the hexagon are respectively provided with a liquid inlet 510 and a liquid outlet 511. When placed vertically, the liquid inlet 510 is located at the lower end of the hexagon, and the liquid outlet 511 is located at the upper end of the hexagon, forming a hydrodynamically dominant shape in which the liquid flows through the reaction chamber in a balanced manner. As shown in Figure 11, a temperature sensor 561 is fixed on the bottom of the sequencing reaction cell 51 through thermally conductive silica gel, and a heating element for accommodating a semiconductor heater 571 is provided on the other side of the reaction cell body 5 where the annular groove 56 is not provided. Chamber 57, the heating chamber 57 is opened on the side of the reaction pool body 5, and a semiconductor heater 571 is fixed on its bottom surface to heat the reaction pool body 5 to ensure that the temperature in the annular groove 56 is controlled at 35±1°C. In this embodiment, the heating chamber 57 is closed by a closing plate 572 so as to be closely attached to the mounting seat 6 of the sequencing chip with the reaction cell body 5 .

As shown in Fig. 4, Fig. 12 and Fig. 13, four third through holes are arranged on the seat body 63 of the mounting base 6 of the sequencing chip, and the mounting base 6 of the sequencing chip passes through the four third through holes passing through the four third through holes. A screw 61 is connected with the four screw holes 612 provided on the back plate 54 of the reaction cell body 5, thereby realizing the fixed connection between the seat body 63 of the mounting seat 6 of the sequencing chip and the back plate 54 of the reaction cell body 5 . As shown in Figures 12-15, the base body 63 of the mounting seat 6 of the sequencing chip is provided with an installation cavity 69 for accommodating the sequencing chip 7, and one end of the installation cavity 69 is provided with a positioning ring for positioning the sequencing chip 7 67, the inner wall of the installation cavity 69 is provided with a positioning groove along its radial direction, and the positioning groove is provided with a positioning groove that slides along the positioning groove and extends into the installation cavity 69 to hold the sequencing chip 7 so that it is attached to the positioning ring 67 or retracted for positioning. The sliding tongue plate 62 of the sequencing chip 7 is loosened in the groove. The distance between the sliding tongue 62 and the positioning ring 67 is exactly equal to the thickness of the sequencing chip when the sliding tongue 62 is stretched out, and the sequencing chip 7 can be tightly attached to the positioning ring 67 when the sliding tongue 62 is stretching out. The positioning grooves are arranged symmetrically on the inner wall of the installation cavity 69 and run through the inner wall. The number of them can be two or four. Evenly distributed in the circumferential direction at the same position. Of course, more positioning grooves can be provided in order to increase the clamping force, but more positioning grooves will cause inconvenience in operation. In this embodiment, there are two positioning grooves, and for the convenience of processing, the positioning grooves are in the shape of round holes. As shown in Figure 5, the size of a section of the positioning groove close to the installation cavity 69 is smaller, forming a second step 623 for limiting the sliding tongue plate 62; that is, the section of the positioning groove close to the installation cavity 69 is a smaller round hole , and the rest is a larger circular hole. As shown in FIGS. 9 and 10 , the sliding tongue 62 is provided with a protruding protrusion 621 . As shown in Figure 7, when the sliding tongue plate 62 extends into the installation cavity 69, the protruding part 621 of the sliding tongue plate 62 abuts against the second step 623, and one end of the sliding tongue plate 62 extends into the installation cavity 69 Hold the sequencing chip 7 with a card. The outer end of the positioning groove is provided with a baffle plate 64 fixedly connected with the mounting base 6, and a second spring 633 is provided between the protrusion 621 of the sliding tongue plate 62 and the baffle plate 64. In this embodiment, the second spring 633 Covered on the other end of the sliding tongue 62 , the two ends of the second spring 633 abut against the protrusion 621 and the baffle 64 respectively. As shown in Figures 13 and 14, an annular groove 66 is formed on the surface of the mount 6 of the sequencing chip, and the annular groove 66 surrounds the outside of the mounting cavity 69, so that the sequencing chip 7 is located in the area surrounded by the annular groove 66 .

As shown in FIG. 8 , the baffle plate 64 is fixedly connected to the mounting base 6 by two screws 41 , which is convenient for installation and disassembly. The second spring 633 is sleeved on the sliding tongue 62 between the protruding portion 621 and the baffle 64, as shown in Figures 11-13, the baffle 64 is provided with the end of the sliding tongue 62 away from the installation cavity 69 to pass through The inner side of the baffle plate 64 is provided with a sleeve 45 with an inner diameter larger than the through hole 44 , and a third step 49 is formed between the sleeve 45 and the through hole 44 for making the end of the second spring 633 lean against it. In this embodiment, the second spring 633 can also be replaced by other second elastic components, and the baffle 64 can also use other components, as long as it can be against the other end of the second spring 633 . When installing the sequencing chip 7, toggle the end of the sliding tongue plate 62 protruding into the installation cavity 69 to retract it into the positioning groove, and the protruding part 621 of the sliding tongue plate 62 squeezes the second spring 633 to push the sequencing chip 7 is put into the installation cavity 69 and pasted on the positioning ring 67, and then the sliding tongue 62 is released. The sliding tongue 62 is stretched out under the action of the second spring 633, and is clamped on the side of the sequencing chip 7, so that the sequencing chip 7 is pasted. It is tight on the positioning ring 67 to ensure that the sequencing chip 7 will not come out even when it is turned upside down.

In order to facilitate the loading and unloading of the sequencing chip 7, preferably, as shown in FIG. 5, the DNA sequencer in this embodiment includes a support frame 84, and the base 8 is connected to the support frame 84 through a rotating shaft 87 as shown in FIGS. 4 and 6. , when installing or removing the sequencing chip 7, let the base 8 be in a horizontal position, when performing a sequencing reaction, the base 8 is in a vertical position as shown in Figure 5, and the base 8 is fixedly connected to the support frame 84 by a screw 841, the screw 841 has a rotary knob on one end and a connecting thread on the other.

As shown in Figure 21, the CCD camera matched with the DNA sequencer of this embodiment includes a camera body 1 and an optical signal located in the camera body 1 for reading the sequencing reaction occurring on the sequencing chip 7. The fiber optic panel 4, one end of the fiber optic panel 4 protrudes from the front end of the camera body 1 and is directly connected to the sequencing chip mounting seat 6 of the DNA sequencer (the structure of the sequencing chip mounting seat 6 is described in detail above) for sequencing One side of the chip 7 is in direct contact to obtain the light signal generated by the sequencing reaction on the other side of the sequencing chip 7 . In this embodiment, preferably, the optical fiber core diameter of the optical fiber panel is 6 μm, and the optical fiber core diameter can be selected within 1-15 μm. The fiber optic panel 4 directly protrudes from the front end of the camera body 1 without using a lens. The fiber optic panel 4 can be directly coupled with the sequencing chip 7 to simplify the optical path, reduce light loss, and improve the quality of the sequencing signal.

As shown in FIG. 21 and FIG. 24 , in this embodiment, preferably, a connecting flange 2 is connected to the front end of the camera body 1 . As shown in Fig. 21, Fig. 22 and Fig. 523, the front end of the camera body 1 has a protruding ring 21 for inserting into the annular groove 66 provided on the mounting seat 6 of the sequencing chip and closely fitting with the annular groove 66. The connecting flange 2 and the camera body 1 are connected by bolts, and the fiber optic panel 4 in the camera body 1 is located in the area surrounded by the collar 21 , that is, the collar 21 protrudes from the connecting flange 2 and surrounds the fiber optic panel 4 . The end face of the fiber optic panel 4 protrudes from the convex ring 21 for a certain distance, which can be adjusted between 1-3mm. The end face of the sequencing chip can extend into the installation cavity 69 of the mounting seat 6 of the sequencing chip and directly contact with the sequencing chip 7 . In this embodiment, by setting the optical fiber panel 4 that can be directly coupled with the sequencing chip, the weak visible light generated by the sequencing reaction on the sequencing chip 7 can be directly received by the optical fiber panel 4 and converted into an electrical signal, thereby improving the optical coupling efficiency. The coupling efficiency can exceed 70%, ensuring the acquisition of high-quality sequencing signals. In addition, the annular groove 66 matches the shape of the collar 21 so that the insert of the collar 21 into the annular groove 66 can form a sealing fit. As shown in Figure 24, when the CCD camera is brought close to the mounting seat 6 of the sequencing chip, the connecting flange 2 installed on the front end of the camera body 1 is close to the sequencing chip 7 on the mounting seat 6 of the sequencing chip, and the ring on the connecting flange 2 The convex ring 21 enters the annular groove 66 on the mounting base 6 of the sequencing chip. The convex ring 21 matches the shape of the annular groove 66 and can be closely matched to form a sealed structure, thereby forming a light-tight darkroom environment to further ensure the sequencing reaction. The generated light energy is received by the fiber optic panel 4 to ensure the acquisition of sequencing signals. By adding a connecting flange 2 with a protruding ring 21 at the front end of the camera body 1, and opening a corresponding annular groove 66 on the mounting seat 6 of the sequencing chip, a dark room is formed between the mounting seat 6 of the sequencing chip and the camera body 1 The environment ensures that when the sequencing chip 7 is coupled with the optical fiber panel 4 located at the front end of the camera body 1, it is free from external light interference, thereby reducing background noise and further ensuring the acquisition of sequencing signals with a high signal-to-noise ratio.

FIG. 24 shows the structure when a reaction chamber of the DNA sequencer used in the control system of this embodiment is connected to a CCD camera, that is, a schematic structural diagram of the DNA sequencer when it is in a working state. As shown in Figure 24, when performing a sequencing reaction, the camera body 1 of the CCD camera is close to the reaction chamber, and the outer end surface of the optical fiber panel 4 exposing the front end of the camera body 1 is pasted on the sequencing chip 7, along with the movement of the camera body 1 to the reaction chamber direction movement, the optical fiber panel 4 pushes the sequencing chip 7 to move, since the sequencing chip 7 is fixed in the mounting seat 6 of the sequencing chip, the mounting seat 6 of the sequencing chip and the reaction pool body 5 are also fixedly connected, so that the sequencing chip 7 can drive The reaction cell body 5 slides on the guide rod 9 along the axial direction of the guide rod 9. Since the reaction cell body 5 is subjected to the force of the first spring 81 between the reaction cell body 5 and the base 8, the outer end surface of the sequencing chip 7 and the optical fiber The panels 4 are close to each other. Before the outer end face of the sequencing chip 7 is completely parallel to the optical fiber panel 4, due to the force of the first spring 81, the reaction cell body 5 is subjected to the first spring 81 during the process of the outer end face of the optical fiber panel 4 being close to the sequencing chip 7. When the force of a spring 81 is exerted, it slightly moves in the radial direction of the guide rod 9, and drives the sequencing chip 7 to move slightly in the radial direction of the guide rod 9, thereby realizing the parallelism between the outer end surface of the optical fiber panel 4 and the sequencing chip 7 Adjustment. The sequencing chip 7 is adjusted in the axial direction of the guide rod 9, so that the outer end surface of the sequencing chip 7 and the outer end surface of the optical fiber panel 4 are as close as possible, and the reaction cell body 5 moves slightly in the radial direction of the guide rod 9, The highest possible parallelism accuracy between the outer end face of the fiber optic panel 4 and the sequencing chip 7 is achieved, and external optical pollution is prevented to the greatest extent when the optical fiber panel 4 reads the fluorescent signal of the sequencing reaction from the sequencing chip 7 , thereby reducing the background noise and realizing the acquisition of sequencing signals with high signal-to-noise ratio.

The structure and working process of the two-dimensionally adjustable support device of the DNA sequencer applied in the control system of this embodiment will be described below with reference to FIG. 1 , FIG. 25 , and FIG. 26 .

As shown in Fig. 1, Fig. 25, and Fig. 26, the two-dimensionally adjustable support device of the DNA sequencer used in the control system of the present embodiment includes a plurality of parallel CCD cameras 110 for driving the CCD camera 110 in the reaction chamber assembly 104. The first linear motion mechanism 31 that switches between the mounting seats of the reaction chamber and the second linear motion mechanism 32 that drives the CCD camera 110 close to or away from each reaction chamber, wherein the first linear motion mechanism 31 includes a first guide rail seat 311, the first guide rail 312 arranged on the first guide rail seat 311, the first slider 313 that moves linearly along the first guide rail 312, and the first ball screw kinematic pair located in the first guide rail seat 311, the first ball screw The nut (not shown in Fig. 25) of the bar movement pair is fixedly connected with the first slide block 313; The second slider 323 of the second guide rail 322 linearly moves and the second ball screw kinematic pair located in the second guide rail seat 321, the nut of the second ball screw kinematic pair (not shown in FIG. 25, see the description below ) is fixedly connected with the second slider 323; the second rail seat 321 is fixedly connected with the first slider 313 and the second rail 322 is vertically arranged with the first rail 312, and the second slider 323 is connected with the CCD camera 110 above it The supporting seat 35 is fixedly connected. Since the second guide rail 322 is perpendicular to the first guide rail 312, the second linear motion mechanism 32 and the CCD camera 110 fixedly connected to it can switch between different reaction chambers when moving along the first guide rail 312. When the linear motion mechanism 32 moves to the mounting seat facing a certain reaction chamber, driven by the second linear motion mechanism 32, the CCD camera 110 moves toward or away from the mounting seat of the reaction chamber. When the CCD camera 110 moves towards the mounting seat of the reaction chamber and is closely attached to the mounting seat, the sequencing work is carried out to capture the optical signal in the reaction chamber. Driven by 31, the mounting seats of different reaction chambers are switched.

The structure of the first linear motion mechanism 31 and the second linear motion mechanism 32 are basically the same, the difference is that the top of the first slider 313 is fixedly connected to the second guide rail seat 321, and the top of the second slider 23 is fixedly connected is the support seat 35 of the CCD camera 110 . The structure and working process of the second linear motion mechanism 32 will be described in detail below in conjunction with Fig. 26-Fig. 31 on.

As shown in Figures 26-29, the second linear motion mechanism 32 also includes a second servo motor 326 connected to the second lead screw 271 of the second ball screw motion pair. Of course, the first linear motion mechanism also includes a The first servo motor (not shown in the figure) connected to the first lead screw of the first ball screw kinematic pair. The second linear motion mechanism 32 realizes linear motion through two second guide rails 322 arranged in parallel on the second guide rail seat 321, and the second slider 323 slides on the second guide rails 322 to ensure the accuracy of its linear motion. The second servo motor 326 drives the second slider 323 to slide on the second guide rail 322 through the second ball screw kinematic pair, wherein the second lead screw 271 of the second ball screw kinematic pair is located between the two second guide rails 322 Between, the nut 272 of the second ball screw kinematic pair is fixedly connected with the second slider 323 , so that the accuracy of the second linear motion mechanism 32 can be ensured. As shown in FIG. 27 and FIG. 28 , the second lead screw 271 of the second ball screw kinematic pair is connected to the second servo motor 326 through a coupling 273 .

As shown in FIGS. 26-29 , a plurality of sensors 38 are provided on one side of the second guide rail seat 321 along the length direction of the second guide rail 322 . The sensor 38 is a photoelectric sensor, and one side of the second guide rail seat 321 is provided with five grooves 380 side by side along the length direction of the second guide rail 322. Set in the length direction, one side of the groove 380 is the light-emitting side, and the other side is the sensing side, and the second slider 323 is fixedly connected with a light-shielding member 381 that can pass through the groove 380, as shown in Figure 26 and Figure 29 , The shading member 381 is an "L" shaped sheet object. When the second slider 323 moves and drives the shade 381 to pass through each groove 380 in turn, the sensing side of the corresponding groove 380 sends a corresponding signal to the control device. The position of the groove 380 in the length direction of the second guide rail 322 is set according to actual needs. In this embodiment, there are five grooves. When piece 381 reaches these two positions, control device just sends alarm signal. The three grooves in the middle correspond to the three normal working positions of the second servo motor. When the light shielding member 381 reaches these three positions, the inductive lateral control device sends out corresponding signals. Of course, a corresponding number of sensors can also be provided on one side of the first guide rail seat 311 according to the number of mounting seats and the need for position limitation.

As shown in Fig. 1 and Fig. 25, one end of the first guide rail seat 311 is provided with a first stop block 319, one end of the second guide rail seat 321 is provided with a second stop block 329, and the other end is provided with a stop bar 328. Two guide rail seats 321 fix the limit seat 281 near one end of the mounting seat, and the limit rod 328 is passed in the limit seat 281 along the length direction of the second guide rail 322, and at one end of the limit rod 328 and the limit seat 281 is provided with the spring that plays buffering effect, when sensor breaks down, when the second slide block 323 can't stop automatically to the direction motion of support frame 84, stop it to continue to advance by limit bar 328 and limit seat 281, avoids The CCD camera 100 collides with the reaction chamber assembly 104 to prevent damage.

The working process of the reagent supply assembly of this embodiment will be described in detail below with reference to FIG. 1 , FIG. 30 and FIG. 31 .

As shown in Figure 30, the reagent supply assembly includes: a preparatory pipeline 710 for accommodating a complete set of reaction reagents to be reacted in sequence, a buffer pipeline 720 for flowing through a buffer, and a reaction chamber for supplying the reaction chamber. The main pipeline 700 of the mixed solution of the reagent and the buffer solution; the preparation pipeline 710 , the buffer pipeline 720 and the main pipeline 700 are connected through a three-way connector 760 . A complete set of reaction reagents refers to all reagents for a single reaction in a reaction chamber.

The first peristaltic pump 701 for extracting sequencing reaction reagents; the first multi-way reversing valve 702 with multiple inlets and one outlet, the second multi-way reversing valve 712 with one inlet and multiple outlets; the first multi-way reversing valve 712 Each inlet of the reversing valve 702 is communicated with a reagent bottle 741 respectively, and the outlet of the first multi-way reversing valve 702 is connected with the preliminary pipeline 710 through the first peristaltic pump 701; it is used to extract buffer solution from the buffer solution bottle 743 The second peristaltic pump 711, the second peristaltic pump 711 is connected with the buffer pipeline 720. The inlet of the second multi-way reversing valve 712 communicates with the main pipeline 700, and an outlet of the second multi-way reversing valve 712 communicates with the waste liquid barrel 742 for bypassing; The outlets are respectively communicated with the liquid inlet of one reaction chamber 750; the liquid outlet of each reaction chamber 750 is communicated with the waste liquid bucket. As shown in Figure 30, the overall liquid circuit relies on the first peristaltic pump 701 and the second peristaltic pump 711 to provide liquid flow pressure. The first peristaltic pump 701 and the second peristaltic pump 711 are respectively driven by corresponding stepping motors. The designed sequence is sequentially selected through the first multi-way reversing valve 702, and the buffer solution is merged and diluted when passing through the three-way connector 760, enters the main pipeline 700, and enters the four reaction chambers respectively through the second multi-way reversing valve 712 One of 750 or the bypass liquid path (the bypass liquid path is directly entering the waste liquid barrel 742 from the second multi-way reversing valve 712 without passing through the reaction chamber 750). As a preference, in this embodiment, the main pipeline 700 is provided with a first vacuum defoamer 731 to reduce dissolved gas and precisely control the amount of reagents and buffers entering the reaction chamber. In order to better eliminate dissolved gas and air bubbles in the liquid path, because the flow rate of the buffer solution is fast, reaching 4mL/min, it is easier to generate air bubbles. Vacuum defoamer 732. The second vacuum defoamer 732 removes the bubbles and most of the dissolved gases present in the buffer. Both the first vacuum defoamer 731 and the second vacuum defoamer 732 are provided with a negative gas pressure through a vacuum pump connected thereto. The first multi-way reversing valve 702 and the second multi-way reversing valve 712 can be electromagnetic valves.

There are ten reagent bottles 741, nine of which are used to contain nine kinds of dCTP, dGTP, dTTP, α-thio dATP, ATP, substrates (fluorescein and APS), apyrase, and apyrase inhibitors for sequencing reactions reagents, and a reagent bottle for buffers used to isolate said reagents. (The buffer solution in this reagent bottle is used to isolate adjacent reagents, which is different from the buffer solution in the buffer solution bottle 743.) The buffer solution bottle 743 holds the buffer solution for dilution and cleaning. Among them, reagents such as dCTP, dGTP, dTTP, dATP of α-position thio, ATP, substrate (fluorescein and APS), apyrase and apyrase inhibitor are respectively connected to the inlet of the first multi-way reversing valve 702. In this embodiment Among them, the first multi-way reversing valve 702 and the second multi-way reversing valve 712 are both tens and eleven-way valves, so as to reserve spare channels. According to the test reaction sequence, the outlet of the first multi-way reversing valve 702 is connected to one of the reagent bottles, driven by the first peristaltic pump 701, the flow rate is 0.7mL/min, and the first peristaltic pump 701 stops for 1s before and after each reagent switch to avoid cross-contamination of reagents.

The buffer solution in the buffer solution bottle 743 is driven by the second peristaltic valve 731 , enters the three-way connector 760 , and enters the main pipeline 700 after being mixed with the reagent and diluted. The pipeline from the outlet of the first peristaltic pump 701 to the inlet of the three-way connector 760, that is, the length of the preliminary pipeline 710 is 285 mm, and a complete set of reagents for a single reaction in a reaction chamber will be arranged in sequence in the preliminary pipeline 710, and then enter the The three-way connector 760, mixed with the buffer solution, is continuously injected into the reaction chamber 750 through the main pipeline 700 and the subsequent liquid path to participate in the sequencing reaction, which avoids the need to stop the second peristaltic pump 711 multiple times in one reaction, and solves the problem of During the process of stopping the pump, the reagents of the previous reaction lack power in the reaction chamber, resulting in the problem of uneven reaction. In this embodiment, the length of the preparatory pipeline 710 can at least accommodate a complete set of reagents for a single reaction in one reaction chamber. The mixed liquid in the main pipeline 700 enters the second multi-way reversing valve 712, and enters the reaction chamber 750 from the liquid inlet port of one of the reaction chambers 750 through program selection, and performs sequencing reaction. The liquid outlet port of the reaction chamber 750 is connected with the waste liquid Barrel 742 communicates. If the cleaning process is performed, the liquid from the second multi-way reversing valve 712 directly enters the bypass liquid path, and the bypass liquid path is directly connected to the waste liquid bucket 742. In this embodiment, the second multi-way reversing valve The four outlets of the valve 712 are communicated with the liquid inlets of the four reaction chambers 750 respectively, and one outlet of the second multi-way reversing valve 712 is communicated with the waste liquid bucket 742 through the bypass liquid path, so as to flush the pipeline with buffer solution, and the other Four outlets are reserved to communicate with the reaction chamber 750 for the reaction reagents to flow out.

In order to achieve better results, the length of the preparation line 710 should be long enough to accommodate a full set of reagents (nine) for a single reaction. As a preferred solution, the length of the preparatory pipeline 710 should also be long enough to accommodate the complete set of reagents for a single reaction and the buffer between different reagents in the complete set of reagents.

The control method of the above-mentioned reagent supply component will be described in detail below in conjunction with the flow chart shown in FIG. 31 , including the following steps:

S1: Turn on the first peristaltic pump;

S2: According to the order of the sequencing reaction, open an inlet of the first multi-way reversing valve, make its outlet communicate with the reagent bottle containing the first reagent required for the sequencing reaction, and extract the reagent bottle The reagents in the bottle enter the preliminary pipeline connected to the outlet of the first peristaltic pump, and the extraction sequence of the reagents is carried out according to the test requirements of the sequencing reaction. Each reagent bottle corresponds to a different reaction reagent, and is reversed with the first multi-channel. One inlet of the valve corresponds to;

S3: When the first reagent reaches the predetermined amount, stop the first peristaltic pump for a period of time, such as 1s, which can be selected between 1-2 seconds to avoid interference between adjacent reagents;

S4: Turn on the first peristaltic pump, open the other inlets of the first multi-way reversing valve in turn, draw other reagents into the preparation pipeline, and stop the first peristaltic pump when each reagent reaches a predetermined amount. A peristaltic pump for a period of time, until a complete set of reagents for a single reaction in a reaction chamber are all sent into the preparation pipeline;

According to the order of the sequencing reaction, extract the second reagent required, then stop the first peristaltic pump, then extract the third reagent, the fourth reagent, ..., and so on, until the required one reaction chamber The complete set of reagents for a single reaction are all fed into the preparation pipeline. In this embodiment, there are nine kinds of reagents in total, and they are nine kinds of reagents for the sequencing reaction in sequence: dCTP, dGTP, dTTP, dATP with α-position thio, ATP , fluorescein, APS, apyrase and apyrase inhibitors. The amount of each reagent, that is, the opening time of the first peristaltic pump is pre-set according to the requirements of the sequencing reaction. The whole set of reagents is arranged on the preparation pipeline in order to realize the cycle of performing a sequencing reaction in one reaction chamber without stopping Any pump, to avoid the lack of power of the front liquid caused by stopping the pump, reducing the flow rate and affecting the quality of the sequencing reaction in the reaction chamber; in order to prevent the two adjacent reagents from interfering with each other, in this embodiment, after completing the extraction of any reagent , turning on the first peristaltic pump, so that the inlet of the first multi-way reversing valve communicates with a reagent bottle containing a buffer (one of which contains a buffer), and is used to draw the buffer to form a A liquid column, which is located between two adjacent reagents to isolate the two adjacent reagents.

S5: Simultaneously turn on the first peristaltic pump and the second peristaltic pump for drawing the buffer, and the complete set of reagents and buffer are connected together through the three-way connection connected with the first peristaltic pump and the second peristaltic pump The device enters the main pipeline;

S6: Open an outlet of the second multi-way reversing valve, the mixed solution of the reagent and the buffer solution in the main pipeline enters the corresponding reaction chamber, and the opening sequence of the second multi-way reversing valve is carried out according to the reaction order of the reaction chamber Setting, in this embodiment, the four reaction chambers are sequentially numbered as the first, second, third, and fourth reaction chambers according to their positions.

According to the above control method, the purpose of providing a complete set of reagents for each reaction chamber in turn is realized.

As a preferred solution, the method for controlling the reagent supply assembly of this embodiment further includes the following steps:

S7: Judging whether a complete set of reagents for a single reaction in a reaction chamber have all entered the reaction chamber, if yes, perform step S8;

S8: The outlet of the second multi-way reversing valve connected to the waste liquid barrel is connected to the inlet of the second multi-way reversing valve, and the inlet of the first multi-way reversing valve is connected to the buffer fluid One of the reagent bottles is connected, and the preparation pipeline and the main pipeline are flushed. After a complete set of reagents for a single reaction in one reaction chamber is supplied, the preparation pipeline and the main pipeline are flushed to prepare for the supply of a complete set of reagents in the next reaction chamber.

In order to prevent interference between adjacent reagents, in this embodiment, the following steps are also added between steps S3 and S4:

S31: Make the outlet of the first multi-way reversing valve connected with air for a period of time, the time is calculated according to the time situation, and selected from 0.5s-1.5s to form the air column in the preparation pipeline The length meets the requirements of isolating adjacent reagents and does not affect the sequencing reaction.

The control system for the DNA sequencer of this embodiment will be described below with reference to FIG. 32 and in combination with FIGS. 1-31 .

As mentioned above, the DNA sequencer in this embodiment includes a plurality of reaction chambers; a CCD camera 110 for collecting the optical signal generated by the DNA sequencing reaction in each reaction chamber; Adjustable supporting device, the supporting device includes a first servo motor (not shown) that drives the CCD camera 110 to switch between different reaction chambers and a second servo motor 326 that drives the CCD camera 110 to approach or move away from one of the reaction chambers, supporting The device is provided with a plurality of sensors 38 for detecting the position of the CCD camera along the direction approaching or away from the reaction chamber; and a reagent supply assembly for providing reaction reagents and buffer solutions for each reaction chamber, the reagent supply assembly includes a The first peristaltic pump 701 for reagents, the second peristaltic pump 711 for pumping buffer, the first multi-way reversing valve 702 for selecting reagents and the second multi-way reversing valve 712 for selecting reaction chambers, the second Both the first multi-way reversing valve 702 and the second multi-way reversing valve 712 are rotary valves driven by motors. That is, the motor drives the spool to rotate, so that one of the inlets of the first multi-way reversing valve 702 is communicated with the outlet. At this time, the reagent bottle connected to the inlet is communicated with the outlet of the first multi-way reversing valve 702. When changing the reaction reagent, the motor drives the rotary spool to rotate until the desired inlet and outlet are connected. The working process of the second multi-way reversing valve 712 is similar to that of the first multi-way reversing valve 702 . The difference is that when the motor rotates, the position of the outlet connected to the inlet of the second multi-way reversing valve 712 is changed, because each reaction chamber is communicated with an outlet of the second multi-way reversing valve 712, thereby changing the connection with the second multi-way reversing valve 712. The multi-way reversing valve 712 communicates with the reaction chamber.

As shown in Figure 32, the control system of this embodiment includes a PLC (programmable logic controller); the PLC is electrically connected to the controller of the first servo motor and the controller of the second servo motor to control the first servo motor and the second servo motor. The start, stop and rotation direction of the servo motor; the PLC is electrically connected with the first peristaltic pump and the second peristaltic pump to control the start, stop and speed of the first peristaltic pump and the second peristaltic pump; the PLC is respectively connected with the first multi-channel The driving motor of the reversing valve is electrically connected with the driving motor of the second multi-way reversing valve to control the on-position of the first multi-way reversing valve and the second multi-way reversing valve: PLC controls the first multi-way reversing valve 702 to select the reagent bottle connected to it, and the PLC controls the connected position of the second multi-way reversing valve 712 to select the reaction chamber connected to it. The PLC is electrically connected to the CCD camera to control the on and off state of the CCD camera; the PLC is also connected to a plurality of sensors (only one sensor is shown in Figure 32, and the other sensors are connected to the PLC) to receive the CCD camera signal sent by the sensor. location signal. When the CCD camera 110 is attached to the reaction chamber and reaches the expected reaction time, the CCD camera is turned on to acquire the light signal generated by the sequencing reaction to form an image.

As shown in FIG. 32 , the control system of this embodiment further includes a host computer connected to the PLC, and the host computer is used to set a control program operated by the PLC and transmit the control program to the PLC. The control program is the working process of PLC, that is, the PLC to the controlled object, including the controller of the first servo motor, the controller of the second servo motor, the first peristaltic pump 701, the second peristaltic pump 711, the first multi-channel Types and sequences of control signals sent by the drive motor of the reversing valve 702 , the drive motor of the second multi-way reversing valve 712 and the CCD camera 110 . In this embodiment, the upper computer performs serial communication with the PLC through the RS232 port. The host computer communicates with the CCD camera through the USB port to receive the image data acquired by the CCD camera.

The temperature control module of the control system for the DNA sequencer according to the embodiment of the present invention will be described below with reference to FIG. 33 and in conjunction with FIG. 11 .

As shown in Figure 33, the temperature control module includes a relay connected to the PLC and a temperature sensor 561 and a semiconductor heater 571 arranged in the reaction chamber; the PLC receives the signal from the temperature sensor 561 and is connected with the primary circuit of the relay, and the secondary circuit of the relay The stage circuit is connected in the power supply circuit of the semiconductor heater 571 . When the temperature in the annular groove 56 was lower than the predetermined temperature; the PLC sent a connection signal to the relay, so that the power supply circuit of the semiconductor heater 571 was connected; when the temperature in the annular groove 56 was higher than another predetermined temperature, the PLC A signal is sent to the relay to disconnect the power supply circuit of the semiconductor heater 571, thereby ensuring that the temperature in the annular groove 56 is controlled within the target temperature range. In this embodiment, the relay is an electromagnetic relay. Figure 33 only shows the composition of the temperature control module in one reaction chamber, and the other reaction chambers are the same. As shown in FIG. 11 , the temperature sensor 561 and the semiconductor heater 571 are arranged in the reaction chamber, and the temperature sensor 561 is fixed on the bottom of the sequencing reaction cell 51 through thermal silica gel, and the reaction cell body 5 is not provided with an annular groove 56 on the other side On one side, there is a heating chamber 57 for accommodating a semiconductor heater 571. The heating chamber 57 is opened on the side of the reaction cell body 5, and the semiconductor heater 571 is fixed on its bottom surface to heat the reaction cell body 5.

In this implementation, the PLC adopts Omron's CP1H-40XDT-D-CH type, and controls the first servo motor and the second servo motor through pulse signals respectively. Both the first servo motor and the second servo motor are stepping motors; the PLC sends out The pulse signal controls the motor speeds of the first peristaltic pump and the second peristaltic pump. Omron CP1H-40XDT-D-CH belongs to the relay output type 40-point PLC, and at the same time introduces 8-point output expansion, which is used for BCD (binary coded decimal) control of the first multi-way directional valve and the second multi-way directional valve , By performing BCD control to realize switching of the on position of the valve, etc. PLC controls the temperature by controlling the power supply circuit of the semiconductor heater by controlling the on and off of the relay. The control system of the present invention enables the reagent supply assembly of a DNA sequencer with multiple reaction chambers to supply reagents and buffers in a timely and accurate manner, and enables the CCD camera to read the optical signal in each reaction chamber in a timely manner, thereby realizing Multiple reactions are carried out at the same time, and multiple samples can be sequenced at the same time, which greatly improves the efficiency of DNA sequencing.

Of course, the above are the preferred embodiments of the present invention. It should be pointed out that for those skilled in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications are also considered Be the protection scope of the present invention.

Claims (5)

1. system that is used for the dna sequencing appearance, said dna sequencing appearance comprises a plurality of reaction warehouses; Be used to gather the CCD camera of the optical signal that dna sequencing reaction produced in each said reaction warehouse; Be used to support said CCD camera can the two dimension adjustment bracing or strutting arrangement; Said bracing or strutting arrangement comprise first servomotor that the driven CCD camera switches between different reaction warehouses with drive said CCD camera near or away from second servomotor of one of them reaction warehouse, said bracing or strutting arrangement upper edge near or be provided with a plurality of transmitters that are used to detect said CCD camera position away from the direction of said reaction warehouse; And be the agent delivery assembly that a plurality of said reaction warehouses provide reaction reagent and damping fluid; Said agent delivery assembly comprises first peristaltic pump, second peristaltic pump that is used to extract damping fluid that are used to extract sequencing reaction reagent, be used for more than first logical reversing valves of selective reagents and be used for the more than second logical reversing valves in selective reaction storehouse, and said more than first lead to reversing valves and more than second leads to reversing valves all for by motor-driven rotary valve;

It is characterized in that said system comprises: PLC;

Said PLC is electrically connected with the startup of controlling said first servomotor and second servomotor respectively, stops and turning direction with the unit of the unit of said first servomotor and second servomotor;

Said PLC respectively with state first peristaltic pump, second peristaltic pump is electrically connected with the startup of controlling said first peristaltic pump and second peristaltic pump, stops and rotating speed;

Said PLC is electrically connected to control the on position of logical reversing valve more than first and more than second logical reversing valves with the drive-motor of said more than first logical reversing valves and the drive-motor of more than second logical reversing valves respectively;

Said PLC is electrically connected with said CCD camera to control the open and close state of said CCD camera;

Said PLC also is connected with said a plurality of transmitters respectively to receive the position signal of the CCD camera that said transmitter sends.

2. system as claimed in claim 1; It is characterized in that; Also comprise temperature control modules; Said temperature control modules comprises the rly. that is connected with said PLC and is arranged at TP and the semiconductor heat booster in the said reaction warehouse, and said PLC receives and is connected from the signal of said TP and with the primary return of said rly., and the secondary loop of said rly. is connected in the current supply circuit of said semiconductor heat booster.

3. system as claimed in claim 1 is characterized in that, also comprises the upper computer that is connected with said PLC, and said upper computer is provided with the sequence of control of said PLC operation, and transmits said sequence of control to said PLC.

4. system as claimed in claim 3 is characterized in that, said upper computer is through RS232 port and said plc communication.

5. system as claimed in claim 3 is characterized in that, said upper computer is communicated by letter with said CCD camera through USB port, is used to receive the view data that said CCD camera is obtained.

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