CN102703314B - Control system for DNA (Deoxyribose Nucleic Acid) sequencer - Google Patents
Control system for DNA (Deoxyribose Nucleic Acid) sequencer Download PDFInfo
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- CN102703314B CN102703314B CN2012101648385A CN201210164838A CN102703314B CN 102703314 B CN102703314 B CN 102703314B CN 2012101648385 A CN2012101648385 A CN 2012101648385A CN 201210164838 A CN201210164838 A CN 201210164838A CN 102703314 B CN102703314 B CN 102703314B
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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
Technical field
The present invention relates to the dna sequencing technical field, be specifically related to a kind of Controlling System for the dna sequencing instrument.
Background technology
In the dna sequencing technical field, tetra-sodium sequencing technologies (pyrosequencing), be a kind of novel enzyme cascade sequencing technologies that was grown up in 1987 by people such as Nyren, its repeatable and accurate performance and Sanger method dna sequencing technology compare favourably, and speed improves greatly.
The tetra-sodium sequencing technologies is by the enzyme cascade chemiluminescence reaction in 4 kinds of enzymatic same reaction systems.The principle of tetra-sodium sequencing technologies is: after primer and the template DNA annealing, under the synergy of archaeal dna polymerase (DNA polymerase), ATP sulfurylase (ATP sulfurylase), luciferase (luciferase) and four kinds of enzymes of apyrase (Apyrase), the polymerization of each dNTP on the primer and the release coupling of an optical signal are got up, by detecting release and the intensity of light, reach the purpose of The real time measure dna sequence dna.The reaction system of tetra-sodium sequencing technologies is made of reaction substrate, strand to be measured, sequencing primer and four kinds of enzymes.Reaction substrate be 5 '-phosphinylidyne sulfuric acid (adenosine-5 '-phosphosulfate, APS) and fluorescein (luciferin).
Take turns in the sequencing reaction at each, only add a kind of deoxynucleotide triphosphoric acid (dNTP) in the reaction system, if it can just with the next base pairing of dna profiling, then can be under the effect of archaeal dna polymerase, add 3 ' end of sequencing primer to, discharge the tetra-sodium (PPi) of a molecule simultaneously.Under the effect of ATP sulfurylase, the PPi of generation can with APS in conjunction with forming ATP, under the catalysis of luciferase, the ATP of generation again can with fluorescein in conjunction with forming oxyluciferin, produce visible light simultaneously.Can obtain a special detected peaks by Weak light detection device and process software, the height of peak value then is directly proportional with the base number that is complementary.If the dNTP that adds can not with the next base pairing of dna profiling, then above-mentionedly instead would not take place, just do not have detected peaks yet.Remaining dNTP and residual a small amount of ATP degrade under the effect of Apyrase in the reaction system.After treating that last round of reaction is finished, add another kind of dNTP, above-mentioned reaction is repeated, namely can read dna sequence dna information accurately according to the peak value figure that obtains.
The integrated operation flow process is described below: the DNA sample by fragmentation after, application is built storehouse reagent and is added joint, strand is caught, be bonded to microballoon, the microemulsion pcr amplification, breakdown of emulsion liquid, acquisition is based upon the DNA library on the microballoon, application add model with the laies such as enzyme of library and sequencing reaction needs to the sequence testing chip with micro reaction pool, sequence testing chip and sequencing reagent are mounted on the main frame, start the order-checking program by the control computer according to module number and position, sequencing reaction is carried out in automatization, the data transmission that produces is to the data analysis computer, finish order-checking back application calculating analysis software and carry out the image processing, sequence is read, mass analysis, work such as sequence assembly finally obtain the sequence information of dna sample.The sequence testing chip that is carved with micro reaction pool carries out machine glazing core layer etching by the fibre faceplate of core diameter 25 μ m thickness 2mm and obtains, and etching depth 40 μ m amount to about 3,000,000 micro reaction pools on the chip, wherein about 1,200,000 micro reaction pools of imaging moiety.The micro reaction pool sequence testing chip is the carrier of sequencing reaction, and the DNA Beads and the various sequencing reaction that are loaded with sequencing template all are arranged in the sequence testing chip that is carved with micro reaction pool with enzyme.
In the order-checking process, at sequence testing chip chemical reaction taking place, produces visible light, by CCD(Charge Couple Device) camera catches the optical signal that sequencing reaction produces, and can obtain needed order-checking information.
The contriver finds the dna sequencing instrument of prior art, has only a reaction warehouse, and an instrument can only conduct an experiment, and working efficiency is not high.In order to increase work efficiency, the contriver has made the dna sequencing instrument with a plurality of reaction warehouses, supplies reaction reagent and damping fluids for a plurality of reaction warehouses in good time and uses a CCD camera to gather the coordination problem of the optical signal in a plurality of reaction warehouses in good time because the existence of a plurality of reaction warehouses has just produced.
Summary of the invention
The technical problem to be solved in the present invention is, a kind of Controlling System be used to the dna sequencing instrument with a plurality of reaction warehouses is provided, control agent delivery assembly provides reaction reagent and the damping fluid of sequencing reaction for each reaction warehouse in good time, and control CCD camera motion is in time to read the optical signal in each reaction warehouse.
For addressing the above problem, the present invention adopts following technical scheme:
A kind of Controlling System for the dna sequencing instrument, described dna sequencing instrument comprises a plurality of reaction warehouses; Be used for to gather the CCD camera of the optical signal that the dna sequencing reaction in each described reaction warehouse produces; Be used for to support the bracing or strutting arrangement of can two dimension adjusting of described CCD camera, described bracing or strutting arrangement comprise first servomotor that the driven CCD camera switches between different reaction warehouses and drive described CCD camera near or away from second servomotor of one of them reaction warehouse, described bracing or strutting arrangement upper edge near or be provided with a plurality of sensors for detection of described CCD camera position away from the direction of described reaction warehouse; And the agent delivery assembly that reaction reagent and damping fluid are provided for a plurality of described reaction warehouses, described agent delivery assembly comprise for first peristaltic pump that extracts sequencing reaction reagent, be used for extracting damping fluid second peristaltic pump, 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, described more than first lead to reversing valves and more than second leads to reversing valves all for by motor-driven rotary valve;
Described Controlling System comprises PLC; Described PLC respectively with the controller of the controller of described first servomotor and second servomotor be electrically connected to control described first servomotor and second servomotor startup, stop and turning direction; Described PLC respectively with state startup that first peristaltic pump, second peristaltic pump be electrically connected to control described first peristaltic pump and second peristaltic pump, stop and rotating speed; Described PLC is electrically connected to control the on position that logical reversing valve and more than second more than first leads to reversing valves with the drive-motor of described more than first logical reversing valves and the drive-motor of more than second logical reversing valves respectively; Described PLC and described CCD camera are electrically connected to control the open and close state of described CCD camera; Described PLC also is connected to receive the position signal of the CCD camera that described sensor sends respectively with described a plurality of sensors.
As preferably, described Controlling System also comprises temperature control modules, described temperature control modules comprises the rly. that is connected with described PLC and the temperature sensor and the semiconductor heat booster that are arranged in the described reaction warehouse, described PLC receives and is connected from the signal of described temperature sensor and with the primary return of described rly., and the secondary loop of described rly. is connected in the current supply circuit of described semiconductor heat booster.
As preferably, described Controlling System also comprises the upper computer that is connected with described PLC, and described upper computer arranges the sequence of control of described PLC operation, and transmits described sequence of control to described PLC.
As preferably, described upper computer is by RS232 port and described plc communication.
As preferably, described upper computer is communicated by letter with described CCD camera by USB port, is used for receiving the view data that described CCD camera obtains.
Controlling System for the dna sequencing instrument of the present invention, the agent delivery assembly that makes the dna sequencing instrument is that a plurality of reaction warehouses are supplied reagent and damping fluid timely and accurately, and can make the CCD camera in time read the interior optical signal of each reaction warehouse, realized that a plurality of reactions carry out simultaneously, thereby can check order to a plurality of samples simultaneously, improve dna sequencing efficient greatly.
Description of drawings
Fig. 1 is the perspective view of the dna sequencing instrument of the Controlling System of employing embodiments of the invention;
Fig. 2 is the synoptic diagram of the reaction warehouse assembly of the dna sequencing instrument of the Controlling System of employing embodiments of the invention;
Fig. 3 is that the F-F of Fig. 2 is to view;
Fig. 4 is the decomposing schematic representation of one of them reaction warehouse in the reaction warehouse assembly shown in Figure 2;
Fig. 5 looks synoptic diagram for the master of the base of reaction warehouse shown in Figure 4;
Fig. 6 looks (broken section) synoptic diagram for the left side of Fig. 5;
Fig. 7 is the structural representation that the reaction tank body of reaction warehouse shown in Figure 4 cooperates with tail rod;
Fig. 8 looks (broken section) synoptic diagram for the left side of Fig. 7;
Fig. 9 is the structural representation of the reaction tank body among Fig. 8;
Figure 10 is the structural representation of the tail rod among Fig. 8;
Figure 11 is that the A-A of Fig. 7 is to sectional drawing;
Figure 12 looks synoptic diagram for the master of the mount pad of reaction warehouse shown in Figure 4;
Figure 13 looks (broken section) synoptic diagram for the left side of Figure 12;
Figure 14 is that the B-B of Figure 12 is to sectional schematic diagram;
Figure 15 is that the C-C of Figure 14 is to synoptic diagram;
Figure 16 is the front view of the slip hyoplastron among Figure 14;
Figure 17 is the left view of Figure 16;
Figure 18 is the perspective view of the baffle plate among Figure 14;
Figure 19 is the side-view of baffle plate shown in Figure 180;
Figure 20 is that the D-D of Figure 19 is to sectional view;
Figure 21 is the structural representation (front end of camera body is connected with joint flange) of the CCD camera of the dna sequencing instrument of the Controlling System of employing embodiments of the invention;
Figure 22 is the front view of the joint flange among Figure 21;
Figure 23 is that the E-E of Figure 22 is to sectional view;
Structural representation (broken section) when Figure 24 is connected with the CCD camera for one of them reaction warehouse of the dna sequencing instrument of the Controlling System that adopts embodiments of the invention;
The perspective view that Figure 25 is connected with the CCD camera for the bracing or strutting arrangement of can two dimension adjusting of the dna sequencing instrument of the Controlling System that adopts embodiments of the invention;
Figure 26 is the perspective view of second straight-line motion mechanism of the bracing or strutting arrangement of can two dimension adjusting among Figure 25;
Figure 27 is the vertical view of second straight-line motion mechanism shown in Figure 26;
Figure 28 be Figure 27 along G-G to sectional view;
Figure 29 be Figure 28 along H-H to sectional view;
Figure 30 is the liquid line structure synoptic diagram of agent delivery assembly of dna sequencing instrument of the Controlling System of embodiments of the invention:
Figure 31 is agent delivery assembly control method schema shown in Figure 30:
Figure 32 is the structural representation of the Controlling System that is used for the dna sequencing instrument of embodiments of the invention:
Figure 33 is the structural representation of the temperature control modules of the Controlling System that is used for the dna sequencing instrument of embodiments of the invention (be example with a reaction warehouse).
Main description of reference numerals
101-brace table 103-vibroshock 102-vibration damper plate
104-reaction warehouse assembly 110-CCD camera 105-can the two-dimentional bracing or strutting arrangement of adjusting
1-camera body 2-joint flange 4-fibre faceplate
5-reaction tank body 6-mount pad 7-sequence testing chip
The protruding circle of 8-base 9-tail rod 21-
41-screw 44-through hole 45-cover
49-the 3rd step 51-sequencing reaction pond 54-backboard
55-boss 56-annular recesses 57-heating chamber
58-sealing-ring 61-screw 62-slip hyoplastron
63-pedestal 64-baffle plate 66-ring groove
67-retaining ring 69-installation cavity 80-cavity
The 81-first spring 82-second through hole 84-bracing frame
85-spring lever 86-spring support 87-rotating shaft
91-connection section 92-sliding part 93-lug boss
The 31-first straight-line motion mechanism 32-second straight-line motion mechanism
35-supporting seat 38-sensor
The 271-second leading screw 272-nut 281-limit base
The 311-first track base 312-first guide rail 313-first slide block
The 319-first limited block 321-second track base 322-second guide rail
The 323-second slide block 326-second servomotor 327-shaft coupling
328-gag lever post 380-groove 381-shading piece
510-fluid inlet 511-liquid outlet 561-temperature sensor
571-semiconductor heat booster 572-closure plate 605-glissile segment
606-construction section 610-first step 612-screw
621-protuberance 623-second step 633-second spring
The 700-main pipe line 701-first peristaltic pump 702-more than first logical reversing valves
The 711-second peristaltic pump 712-more than the second logical reversing valve 731-first froth in vacuum devices
The 732-second froth in vacuum device 741-reagent bottle 742-waste liquid barrel
743 damping fluid bottles, 750 reaction warehouse 760-three-way connectors
The 841-screw
Embodiment
Below in conjunction with the drawings and specific embodiments the present invention is described in further detail, but not as a limitation of the invention.
1-Figure 29 describes the detailed construction of the dna sequencing instrument of the Controlling System that adopts embodiments of the invention in detail at first by reference to the accompanying drawings.The Controlling System of present embodiment can be applied in the dna sequencing instrument with a plurality of reaction warehouses, is not limited to the described dna sequencing instrument of present embodiment.
As Fig. 1, Fig. 2 and shown in Figure 3, the applied dna sequencing instrument of Controlling System of the present invention comprises brace table 101, a plurality of vibroshock 103 and the vibration damper plate 102 that is connected with brace table 101 by a plurality of vibroshocks 103; Be used for carrying out the reaction warehouse assembly 104 of dna sequencing reaction, reaction warehouse assembly 104 comprises the bracing frame 84 that vertically is arranged on the vibration damper plate and a plurality of reaction warehouses that are set up in parallel on bracing frame 84; In the present embodiment, be that example describes with four reaction warehouses.Be used for to gather the CCD camera 110 of the optical signal that the dna sequencing reaction in the reaction warehouse produces; Be used for to support CCD camera 110 and drive CCD camera 110 when aiming at one of them reaction warehouse near or away from every reaction warehouse, and and the position that aligns of each reaction warehouse between the bracing or strutting arrangement of can two dimension adjusting 105 that switches; Be used to reaction warehouse assembly 104 that the Controlling System 109 of reaction reagent and damping fluid is provided; Reaction warehouse assembly 104 and the bracing or strutting arrangement of can two dimension adjusting 105 all are fixedly installed on the vibration damper plate 102, and Controlling System 109 is arranged on the brace table 101.In the present embodiment, brace table 101 is positioned on the cabinet (not shown), instrument and circuit element that sequencing analysis is used are set in the cabinet, vibroshock 103 adopts the air damp type vibroshock, reached the isolation external shock, eliminate or weaken it to the influence of dna sequencing instrument work quality, in the present embodiment, respectively settled a vibroshock 103 for four of 102 lower ends jiaos at the bottom of the damping.
Detailed construction and the working process thereof of reaction warehouse assembly 104 of the applied dna sequencing instrument of Controlling System of present embodiment at first is described in conjunction with Fig. 1-Figure 24.
As Fig. 2, Fig. 3, Fig. 4 and shown in Figure 24, in the present embodiment, each reaction warehouse includes the base 8 that is rotatably connected on the bracing frame 84, the reaction tank body 5 that carries out sequencing reaction and is used for installing the mount pad 6 of the sequence testing chip of sequence testing chip 7; The mount pad 6 of sequence testing chip is fixedly connected on the reaction tank body 5, base 8 and reaction tank body 5 are provided with and pass its two a plurality of tail rods 9, and tail rod 9 is fixedlyed connected with base 8, reaction tank body 5 is enclosed within on the tail rod 9, axially sliding and the fine motion that can make progress in the footpath of tail rod 9 of tail rod 9.Be provided with first elastomeric element between reaction tank body 5 and the base 8, as shown in Figure 1, in the present embodiment, first elastomeric element is preferably first spring 81, spring has simple in structure, and the advantage of being convenient to install can also select for use other elastomeric elements that can realize said function to realize certainly.
As Fig. 4, Fig. 5 and shown in Figure 6, a side of base 8 is provided with cavity 80, is provided with two spring levers 85 in the cavity 80 side by side, and spring lever 85 integral body are positioned at cavity 80, can not stretch out cavity 80.Sheathed first spring, 81, the first springs 81 of branch stretch out cavity 80 on each spring lever 85.As preferably, in the present embodiment, the bottom of cavity 80 is provided with spring support 86, and spring lever 85 is positioned on the spring support 86, and the spring support 86 of increase can reduce the required length of first spring 81.It is for the ease of miscellaneous part is installed that cavity 80 is set.
As Fig. 4, Fig. 7 and shown in Figure 11, reaction tank body 5 comprises backboard 54 and is located on the described backboard 54 and boss 55 that a side of toward back plate 54 is protruded, the end face of described boss 55 is provided with the annular recesses 56 of sealing, and the zone that the annular recesses 56 of sealing surrounds is sequencing reaction pond 51; As shown in Figure 7, be embedded with sealing-ring 58 in the annular recesses 56, sealing-ring 58 protrudes the end face of boss 55, and to surround sequencing reaction pond 51, wherein the end face of boss 55 is smooth as much as possible.As Fig. 4, Fig. 7, Fig. 8 and shown in Figure 9, the side that reaction tank body 5 is provided with backboard 54 is attached on the base 8, the boss 55 of reaction tank body 5 is provided with four first through holes, described first through hole is step-like, one section near base is glissile segment 605, another section of first through hole is construction section 606, and the internal diameter of construction section 606 forms first step 610 greater than glissile segment 605.As Fig. 7-shown in Figure 10, one end of tail rod 9 is the connection section 91 that is connected with base 8, the other end of tail rod 9 is provided be used to the lug boss 93 that is held in first step 610 places, and the middle part of tail rod 9 is near the sliding part 92 of part for being used for cooperating with glissile segment 605 of lug boss 93.As shown in figure 12, connection section 91 is provided with screw thread, and as shown in Figure 7, the end of the lug boss 93 of tail rod 9 is provided with for the cutter groove that cooperates screwdriver.As shown in Figure 1, tail rod 9 passes first through hole on the reaction tank body 5 and second through hole 82 on the base 8, and the back that makes connection section 91 expose base 8 is fastening by nut then, thereby realizes fixedlying connected between tail rod 9 and the base 8.Realize being slidingly connected in order to make between reaction tank body 5 and the tail rod 9, the slightly larger in diameter of glissile segment 605 is in the diameter of sliding part 92, like this reaction tank body 5 both can be implemented in tail rod 9 axially on slip, the fine motion that again can realization response pond body 5 makes progress in the footpath of tail rod 9.In the present embodiment, as preferred version, tail rod 9 is provided with lug boss 93, so that when being slidingly connected at reaction tank body 5 on the tail rod 9, by the position-limiting action of lug boss 93 with first step 610, tighten with connection section 91 on during the nut of thread fit, lug boss 93 driving a reaction pond bodies 5 move to the direction of base 8, to compress first spring 81, make between reaction tank body 5 and the base 8 and have certain preload, it is exactly in order to be attached on the base 8 when reaction tank body 5 that above-mentioned first spring 81 stretches out cavity 80, and first spring 81 can be to the certain reactive force of reaction tank body 5.
As Fig. 4, Figure 12, Figure 13 and shown in Figure 14, the mount pad 6 of sequence testing chip comprises pedestal 63 and the installation cavity 69 that is opened on the pedestal 63, the mount pad 6 of sequence testing chip is positioned at the side that reaction tank body 5 is provided with boss 55, fixedlying connected between pedestal 63 and the backboard 54 stretches in the installation cavity 69 boss 55, sequence testing chip 7 is fixed in the outer end of installation cavity 69, be provided with the gap of flowing through with reagent for order-checking between the end face of sequence testing chip 7 and boss 55, this gap is by being embedded at sealing-ring 58 sealings in the annular recesses 56, the size in described gap is about 0.2mm, and sealing-ring 58 convex hole connected in stars 56 contact to surround the sequencing reaction pond 51 for sequencing reaction of sealing with sequence testing chip 7.
As Fig. 7, shown in Figure 11, in the present embodiment, this annular recesses 56 is hexagon, hexagonal two ends have fluid inlet 510 and liquid outlet 511, when reaction tank body 5 is vertically placed, fluid inlet 510 is in hexagonal lower end, and liquid outlet 511 is in hexagonal upper end, forms the liquid stream hydrokinetics advantageous shape that balanced flow is crossed in reaction warehouse.As shown in figure 11, fixedly there is temperature sensor 561 bottom in sequencing reaction pond 51 by heat conductive silica gel, be not provided with the opposite side of annular recesses 56 at reaction tank body 5, offer the heating chamber 57 for ccontaining semiconductor heat booster 571, heating chamber 57 is opened on the side of reaction tank body 5, fixing semiconductor heat booster 571 on its bottom surface with to 5 heating of reaction tank body, guarantees that the temperature that is positioned at annular recesses 56 controls at 35 ± 1 ℃.In the present embodiment, heating chamber 57 is by closure plate 572 sealings, in order to be adjacent on the mount pad 6 of sequence testing chip with reaction tank body 5.
As Fig. 4, Figure 12 and shown in Figure 13, the pedestal 63 of the mount pad 6 of sequence testing chip is provided with four third through-holes, four set on the backboard 54 of four screws 61 of the mount pad 6 of sequence testing chip by passing these four third through-holes and reaction tank body 5 screws 612 are connected, thereby realize fixedlying connected between the backboard 54 of pedestal 63 and reaction tank body 5 of mount pad 6 of sequence testing chip.As Figure 12-shown in Figure 15, the pedestal 63 of the mount pad 6 of sequence testing chip is provided with the installation cavity 69 that comprises for ccontaining sequence testing chip 7, one end of installation cavity 69 is provided with the retaining ring 67 for location sequence testing chip 7, it radially is provided with locating slot the inwall upper edge of installation cavity 69, be provided with in the locating slot along locating slot slide stretch in the installation cavity 69 with hold sequence testing chip 7 make it be attached on the retaining ring 67 or the withdrawal locating slot in to unclamp the slip hyoplastron 62 of sequence testing chip 7.When slip hyoplastron 62 stretches out and the distance between the retaining ring 67 equal the thickness of sequence testing chip just, sequence testing chip 7 is adjacent on retaining ring 67.The locating slot symmetry is located on the inwall of installation cavity 69 and connects its inwall, and its quantity can be even numbers such as two or four, and in order stable holding to live sequence testing chip 7, a plurality of locating slots evenly distribute along the circumferential direction on the axial same position of installation cavity 69.Certainly, in order to improve holding force more locating slots can also be set, but more locating slot can cause the inconvenience of operation.In the present embodiment, locating slot is two, and for easy to process, locating slot is circle hole shape.As shown in Figure 5, locating slot is less near one section size of installation cavity 69, is formed for the second step 623 of spacing slip hyoplastron 62; One section that is the close installation cavity 69 of locating slot is a less circular hole, and rest part is a bigger circular hole.As Fig. 9 and shown in Figure 10, slip hyoplastron 62 is provided with the protuberance 621 of protrusion.As shown in Figure 7, when slip hyoplastron 62 stretched into to installation cavity 69, the protuberance 621 of slip hyoplastron 62 was resisted against described second step 623 places, and an end of slip hyoplastron 62 stretches in the installation cavity 69 to hold sequence testing chip 7.The outer end of locating slot is provided with the baffle plate 64 of fixedlying connected with mount pad 6, between the protuberance 621 of slip hyoplastron 62 and baffle plate 64, be provided with second spring 633, in the present embodiment, second spring 633 is enclosed within the other end of slip hyoplastron 62, and the two ends of second spring 633 are resisted against respectively on protuberance 621 and the baffle plate 64.As Figure 13, shown in Figure 14, have ring groove 66 on the surface of the mount pad 6 of sequence testing chip, ring groove 66 is centered around the outside of installation cavity 69, makes sequence testing chip 7 be positioned at the zone that ring groove 66 surrounds.
As shown in Figure 8, baffle plate 64 is fixedlyed connected with mount pad 6 by two screws 41, is convenient to installation and removal.Second spring 633 is enclosed within on the slip hyoplastron 62 between protuberance 621 and the baffle plate 64, as Figure 11-shown in Figure 13, baffle plate 64 is provided with the through hole 44 that passes away from an end of installation cavity 69 for slip hyoplastron 62, the inboard of baffle plate 64 is provided with internal diameter greater than the cover 45 of through hole 44, the end that is formed for making second spring 633 between cover 45 and the through hole 44 against the 3rd step 49.In the present embodiment, second spring 633 can also adopt other second elastomeric elements to replace, and baffle plate 64 can also adopt miscellaneous part, as long as can be against the other end of second spring 633.When sequence testing chip 7 is installed, stir the end in the installation cavity 69 of extending into of slip hyoplastron 62, make in its withdrawal locating slot, protuberance 621 extruding second spring 633 of slip hyoplastron 62, unclamp slip hyoplastron 62 after sequence testing chip 7 put into installation cavity 69 and be attached on the retaining ring 67, slip hyoplastron 62 stretches out under the effect of second spring 633, is clamped in the side of sequence testing chip 7, sequence testing chip 7 is adjacent on retaining ring 67, even also unlikelyly when guaranteeing that sequence testing chip 7 is inverted deviate from.
For the ease of loading and unloading sequence testing chip 7, as preferably, as shown in Figure 5, the dna sequencing instrument in the present embodiment comprises bracing frame 84, base 8 is by being rotatably connected on the bracing frame 84 as Fig. 4 and rotating shaft 87 shown in Figure 6, when installing or unloading sequence testing chip 7, allow base 8 be positioned at level attitude, when carrying out sequencing reaction, base 8 is positioned at vertical position as shown in Figure 5, base 8 is fixedlyed connected with bracing frame 84 by screw 841, an end band knob of screw 841, and the other end is provided with connecting thread.
As shown in figure 21, comprise camera body 1 with the CCD camera of the dna sequencing instrument of present embodiment coupling and be positioned at the fibre faceplate 4 that is used for reading the optical signal that occurs in the sequencing reaction on the described sequence testing chip 7 of described camera body 1 that the structure that an end of fibre faceplate 4 stretches out the mount pad 6 of the direct mount pad 6(sequence testing chip with the sequence testing chip that is positioned at the dna sequencing instrument of the front end of described camera body 1 sees above and is described in detail) a side of sequence testing chip 7 directly contact to obtain the optical signal that the sequencing reaction on the opposite side that occurs in sequence testing chip 7 produces.In the present embodiment, as preferably, the optical fiber core diameter of described fibre faceplate is 6 μ m, and the optical fiber core diameter can be selected in 1-15 μ m.Fibre faceplate 4 directly stretches out the front end of camera body 1, does not use camera lens, and fibre faceplate 4 can directly be coupled with sequence testing chip 7, simplifies light path, reduces light loss, improves the order-checking quality of signals.
As Figure 21, shown in Figure 24, in the present embodiment, as preferably, the front end of camera body 1 is connected with joint flange 2.Shown in Figure 21, Figure 22 Figure 52 3, the front end of camera body 1 have for insertion be located on the mount pad 6 of sequence testing chip ring groove 66 and with ring groove 66 close-fitting protruding circles 21.Joint flange 2 and camera body 1 are connected by bolt, and the fibre faceplate 4 in the camera body 1 is positioned at the zone that protruding circle 21 surrounds, and namely protruding circle 21 stretches out the outside that joint flange 2 is centered around fibre faceplate 4.The end face of fibre faceplate 4 stretches out protruding circle 21 certain distances, can between 1-3mm, adjust, in the present embodiment, the distance that the end face of fibre faceplate 4 stretches out described protruding circle 21 is 2mm, directly contacts with sequence testing chip 7 so that the end face of fibre faceplate 4 can stretch in the installation cavity 69 of mount pad 6 of sequence testing chip.Present embodiment by arrange can with the direct-coupled fibre faceplate 4 of sequence testing chip, the faint visible light that the sequencing reaction that carries out at sequence testing chip 7 is produced is directly received by fibre faceplate 4, be converted into electrical signal, improve coupling efficiency, coupling efficiency can surpass 70%, has guaranteed obtaining of high-quality order-checking signal.In addition, ring groove 66 is complementary with the shape of protruding circle 21, makes protruding circle 21 insert can form in the ring groove 66 and is sealed and matched.As shown in figure 24, when the mount pad 6 of the close sequence testing chip of CCD camera, be installed in the joint flange 2 of camera body 1 front end near the sequence testing chip 7 on the mount pad 6 of sequence testing chip, the protruding circle 21 of ring-type on the joint flange 2 enters in the ring groove 66 on the mount pad 6 of sequence testing chip, protruding circle 21 is complementary with the shape of ring groove 66, the formation sealed structure can closely cooperate, thereby form the darkroom environment of light stopping property, guarantee that further the luminous energy that sequencing reaction produces is received by fibre faceplate 4, guarantee obtaining of order-checking signal.By have the joint flange 2 of protruding circle 21 in camera body 1 front end increase, and offer corresponding ring groove 66 at the mount pad 6 of sequence testing chip, thereby between the mount pad 6 of sequence testing chip and camera body 1, form the darkroom environment, when guaranteeing fibre faceplate 4 couplings of sequence testing chip 7 and the front end that is positioned at camera body 1, not disturbed by extraneous light, thereby reduced ground unrest, further guaranteed the obtaining of order-checking signal of high s/n ratio.
Structure when the reaction warehouse that Figure 24 shows the applied dna sequencing instrument of Controlling System of present embodiment is connected with the CCD camera, the structural representation when namely the dna sequencing instrument is in running order.As shown in figure 24, when carrying out sequencing reaction, the camera body 1 of CCD camera is near reaction warehouse, the outer face of exposing the fibre faceplate 4 of camera body 1 front end is attached on the sequence testing chip 7, along with the direction of camera body 1 to reaction warehouse moves, fibre faceplate 4 promotes sequence testing chip 7 and moves, because sequence testing chip 7 is fixed in the mount pad 6 of sequence testing chip, also be fixedly connected between the mount pad 6 of sequence testing chip and the reaction tank body 5, thereby sequence testing chip 7 can drive reaction tank body 5 endwisely slipping at tail rod 9 upper edge tail rods 9, because reaction tank body 5 is subjected to the reactive force of first spring 81 between reaction tank body 5 and the base 8, is close to mutually between the outer face of sequence testing chip 7 and the fibre faceplate 4.In the outer face of sequence testing chip 7 with fibre faceplate 4 do not realize fully parallel before, owing to be subjected to the reactive force of first spring 81, be close in the process of sequence testing chip 7 in the outer face of fibre faceplate 4, reaction tank body 5 is when being subjected to the reactive force of first spring 81, fine motion makes progress in the footpath of tail rod 9, drive sequence testing chip 7 fine motion that makes progress in the footpath of tail rod 9, thereby realize the outer face of fibre faceplate 4 and the adjustment of the parallelism between the sequence testing chip 7.Sequence testing chip 7 is by in the adjustment of tail rod 9 on axially, realized pressing close to as much as possible between the outer face of the outer face of sequence testing chip 7 and fibre faceplate 4, reaction tank body 5 fine motion that makes progress in the footpath of tail rod 9, realized having the high parallelism precision of trying one's best between the outer face of fibre faceplate 4 and the sequence testing chip 7, guarantee to greatest extent when fibre faceplate 4 reads the fluorescent signal of sequencing reaction from sequence testing chip 7, prevented external optical pollution, thereby reduced ground unrest, realized obtaining of high s/n ratio order-checking signal.
Structure and the working process thereof of the bracing or strutting arrangement of can two dimension adjusting of the applied dna sequencing instrument of Controlling System of present embodiment is described below in conjunction with Fig. 1, Figure 25, Figure 26.
As Fig. 1, Figure 25, shown in Figure 26, the bracing or strutting arrangement of can two dimension adjusting of the applied dna sequencing instrument of the Controlling System of present embodiment, comprise for drive first straight-line motion mechanism 31 that CCD camera 110 switches between the mount pad of a plurality of reaction warehouses arranged side by side of reaction warehouse assembly 104 and drive CCD camera 110 near or away from second straight-line motion mechanism 32 of each reaction warehouse, wherein, first straight-line motion mechanism 31 comprises first track base 311, be located at first guide rail 312 on first track base 311, along first guide rail, 312 straight-line first slide blocks 313 and the first ball-screw kinematic pair that is positioned at first track base 311, the nut of the first ball-screw kinematic pair (not shown among Figure 25) is fixedlyed connected with first slide block 313; Second straight-line motion mechanism 32 comprises second track base 321, be located at second guide rail 322 on second track base 321, along second guide rail, 322 straight-line second slide blocks 323 be positioned at the second ball-screw kinematic pair of second track base 321, the nut of the second ball-screw kinematic pair (not shown among Figure 25, the description that sees below) is fixedlyed connected with second slide block 323; Second track base 321 is fixedlyed connected with first slide block 313 and second guide rail 322 and the 312 vertical settings of first guide rail, and second slide block 323 is fixedlyed connected with the supporting base 35 of the CCD camera 110 of the side of being located thereon.Because second guide rail 322 and the 312 vertical settings of first guide rail, second straight-line motion mechanism 32 and the CCD camera 110 of fixedlying connected with it are when moving along first guide rail 312, just can between the differential responses storehouse, switch, when second straight-line motion mechanism 32 moves to mount pad over against some reaction warehouses, under the drive of second straight-line motion mechanism 32, CCD camera 110 just towards or away from the direction motion of the mount pad of this reaction warehouse.The mount pad in CCD camera 110 orientating reaction storehouses motion and when being adjacent to mount pad, carry out examining order, optical signal in the catching reaction storehouse, CCD camera 110 is during away from the mount pad of reaction warehouse, just can under the drive of first straight-line motion mechanism 31, between the mount pad of different reaction warehouses, switch.
First straight-line motion mechanism 31 and second straight-line motion mechanism, 32 structures are basic identical, and different is, what the upper fixed of first slide block 313 connected is second track base 321, are the supporting bases 35 of CCD camera 110 and the upper fixed of second slide block 23 connects.Describe structure and the working process of second straight-line motion mechanism 32 in detail below in conjunction with Figure 26-Figure 29, need to prove, to any preferred implementation of second straight-line motion mechanism 32, equally also can be applied on first straight-line motion mechanism 31.
As Figure 26-shown in Figure 29, second straight-line motion mechanism 32 also comprises second servomotor 326 that is connected with second leading screw 271 of the second ball-screw kinematic pair, certainly, first straight-line motion mechanism also comprises the first servomotor (not shown) that is connected with first leading screw of the first ball-screw kinematic pair.Second straight-line motion mechanism 32 is realized straight-line by two second guide rails 322 that are set in parallel on second track base 321, second slide block 323 slides at second guide rail 322 and has guaranteed its straight-line precision, second servomotor 326 drives the slip of second slide block 323 on second guide rail 322 by the second ball-screw kinematic pair, wherein, second leading screw 271 of the second ball-screw kinematic pair is between two second guide rails 322, the nut 272 of the second ball-screw kinematic pair is fixedlyed connected with second slide block 323, can guarantee the precision of second straight-line motion mechanism 32 like this.As Figure 27, shown in Figure 28, be connected by shaft coupling 273 between second leading screw 271 of the second ball-screw kinematic pair and second servomotor 326.
Shown in Figure 26-29, a side of second track base 321 is provided with a plurality of sensors 38 along the length direction of second guide rail 322.Sensor 38 is photo-sensor, one side of second track base 321 is provided with five grooves 380 side by side along the length direction of second guide rail 322, the opening of groove 380 upwards, the length direction of groove 380 is along the length direction setting of second guide rail 322, one side of groove 380 is emission side, and opposite side is fixedly connected with the shading piece 381 that can pass through for the induction side from groove 380 on second slide block 323, as Figure 26, shown in Figure 29, shading piece 381 is one " L " shape board.When 323 motions of second slide block, when drive shading piece 381 passed through each groove 380 successively, the induction side of corresponding groove 380 was just sent corresponding signal to control device.The position of groove 380 on second guide rail, 322 length directions is according to the actual needs setting, be provided with five grooves in the present embodiment, the groove at two ends respectively corresponding allow to leave or near mount pad farthest and closest range, when shading piece 381 reached these two positions, control device just sent guard signal.Three positions of the middle corresponding second servomotor works better of three grooves, when shading piece 381 reached these three positions, induction side direction control device sent corresponding signal.Certainly, a side of first track base 311 sensor that can respective numbers also be set according to quantity and the spacing needs of mount pad.
As Fig. 1, shown in Figure 25, one end of first track base 311 is provided with first limited block 319, one end of second track base 321 is provided with second limited block 329, the other end is provided with gag lever post 328, second track base 321 fixedly has limit base 281 near an end of mount pad, gag lever post 328 is located in the limit base 281 along the length direction of second guide rail 322, and between an end of gag lever post 328 and limit base 281, be provided with the spring of shock absorption, when sensor breaks down, when second slide block 323 can not stop automatically to the motion of the direction of bracing frame 84, stop it to move on by gag lever post 328 and limit base 281, avoid CCD camera 100 and 104 collisions of reaction warehouse assembly, prevent from damaging.
Describe the working process of the agent delivery assembly of present embodiment in detail below in conjunction with Fig. 1, Figure 30 and Figure 31.
As shown in figure 30, the agent delivery assembly comprises: be used for the buffering pipe 720 of the preparation pipeline 710 of a complete set of reaction reagent of the ccontaining question response of arranging in order, the damping fluid that is used for flowing through and be used to the main pipe line 700 of the mixed solution of described reaction warehouse supply reaction reagent and damping fluid; Described preparation pipeline 710, buffering pipe 720 are connected by a three-way connector 760 with main pipe line 700.A complete set of reaction reagent refers to all reagent that carry out the single reaction in the reaction warehouse.
Be used for extracting first peristaltic pump 701 of sequencing reaction reagent; Have more than first logical reversing valves 702 of a plurality of imports and an outlet, have more than second of an import and a plurality of outlets and lead to reversing valves 712; Each import of logical reversing valve 702 more than first is communicated with a reagent bottle 741 respectively, and the outlet of logical reversing valve 702 more than first is connected with preparation pipeline 710 by first peristaltic pump 701; Be used for being connected with buffering pipe 720 from second peristaltic pump, 711, the second peristaltic pumps 711 that damping fluid bottle 743 extracts damping fluid.The import of logical reversing valve 712 more than second is communicated with main pipe line 700, and an outlet of logical reversing valve 712 more than second is communicated with waste liquid barrel 742, is used for bypass; The outlet of other of logical reversing valve 712 more than second is communicated with the fluid inlet of a reaction warehouse 750 respectively; The liquid outlet of each reaction warehouse 750 is communicated with waste liquid barrel.As shown in figure 30, whole liquid road relies on first peristaltic pump 701 and second peristaltic pump 711 that flow stream pressure is provided, first peristaltic pump 701 and second peristaltic pump 711 are driven by corresponding stepper-motor respectively, different reagent are selected by more than first logical reversing valves 702 successively according to the sequential of programdesign, by three-way connector 760 time, converge dilution with damping fluid, enter main pipe line 700, enter one of four reaction warehouses 750 or bypass liquid road (bypass liquid road is exactly directly to enter waste liquid barrel 742 from more than second logical reversing valves 712 without reaction warehouse 750) respectively by more than second logical reversing valves 712.As preferably, in the present embodiment, main pipe line 700 is provided with the first froth in vacuum device 731, and to subdue dissolved gases, accurately control enters reagent in the reaction warehouse and the amount of damping fluid.In order to eliminate gas dissolved and bubble in the liquid road better, because the flow velocity of damping fluid is very fast, reach 4mL/min, easier generation bubble, the pipeline between damping fluid bottle 743 and second peristaltic pump 711 is provided with the second froth in vacuum device 732.The second froth in vacuum device 732 is removed bubble and the most of gas dissolved that exists in the damping fluid.The first froth in vacuum device 731 and the second froth in vacuum device 732 all provide the gas negative pressure by connected vacuum pump.Logical reversing valve 702 more than first and more than second logical reversing valves 712 can adopt magnetic valve.
Reagent bottle 741 has ten, nine kinds of reagent such as nine dATP, ATP that are used for holding the dCTP, the dGTP that carry out sequencing reaction, dTTP, α position sulfo-, substrate (fluorescein and APS), apyrase, apyrase inhibitor wherein also have a reagent bottle to be used for holding damping fluid for isolating described reagent.(damping fluid in this reagent bottle be used for to be isolated adjacent reagent, and is different with the effect of damping fluid in the damping fluid bottle 743), holds dilution in the damping fluid bottle 743 and clean the damping fluid of usefulness.Wherein, reagent such as dATP, the ATP of dCTP, dGTP, dTTP, α position sulfo-, substrate (fluorescein and APS), apyrase and apyrase inhibitor are connected with the import of more than first logical reversing valves 702 respectively, in the present embodiment, logical reversing valve 702 more than first and more than second logical reversing valves 712 are ten 11 logical valves, in order to reserve standby passage.According to test reaction sequential, the outlet of logical reversing valve 702 more than first is communicated with one of them reagent bottle, driven by first peristaltic pump 701, and flow velocity 0.7mL/min, front and back first peristaltic pump 701 that at every turn switches reagent all stops 1s, to avoid the crossed contamination of reagent.
Damping fluid in the damping fluid bottle 743 is driven by second peristaltic valve 731, enters three-way connector 760, goes into main pipe line 700 with the reagent mix dilution is laggard.The pipeline that is exported to three-way connector 760 entrances of first peristaltic pump 701, the length of namely preparing pipeline 710 is 285mm, carrying out a complete set of reagent of single reaction in reaction warehouse will arrange in preparation pipeline 710 according to the order of sequence, enter three-way connector 760 then successively, with inject reaction warehouse 750 continuously through main pipe line 700 and follow-up liquid road after damping fluid mixes and participate in sequencing reaction, avoided in primary first-order equation, needing repeatedly to stop the situation of second peristaltic pump 711, solved because the reagent of secondary response lacks power and causes the problem of reacting inhomogeneous before in the termination of pumping process in reaction warehouse.In the present embodiment, the length of preparation pipeline 710 can be carried out a complete set of reagent that single reacts in the ccontaining reaction warehouse at least.Mixed solution in the main pipe line 700 enters more than second logical reversing valve 712, and enters reaction warehouse 750 by program selection from the fluid inlet of one of them reaction warehouse 750, carries out sequencing reaction, and the liquid outlet of reaction warehouse 750 is communicated with waste liquid barrel 742.If what carry out is matting, then the liquid that comes out from more than second logical reversing valves 712 directly enters bypass liquid road, waste liquid barrel 742 is directly connected on bypass liquid road, in the present embodiment, four outlets of logical reversing valve 712 more than second are communicated with the fluid inlet of four reaction warehouses 750 respectively, and an outlet of logical reversing valve 712 more than second is communicated with waste liquid barrel 742 by bypass liquid road, in order to use the damping fluid flushing pipe, other four outlets are reserved with reaction warehouse 750 and are communicated with, and flow out for reaction reagent.
In order to realize effect preferably, the length of preparation pipeline 710 should be sufficiently long to a complete set of reagent (nine kinds) that can hold the single reaction.As preferred version, the length of preparation pipeline 710 also should be sufficiently long to the damping fluid between the different reagent in a complete set of reagent that holds single reaction and a complete set of reagent.
Below in conjunction with schema shown in Figure 31, describe the control method of above-mentioned agent delivery assembly in detail, comprise the steps:
S1: open described first peristaltic pump;
S2: according to the order of sequencing reaction, open an import of described more than first logical reversing valves, the described reagent bottle of its outlet first kind reagent required with being contained with sequencing reaction is communicated with, the reagent that extracts in the described reagent bottle enters the preparation pipeline that is connected with the described first wriggling pump outlet, the extraction order of reagent is carried out according to the test requirements document of sequencing reaction, the reaction reagent that each reagent bottle is corresponding different, and corresponding with an import of described more than first logical reversing valves;
S3: when first kind of reagent reaches predetermined amount, stopped for first peristaltic pump for some time, as 1s, can select between second at 1-2, avoid the interference between the adjacent reagent;
S4: open described first peristaltic pump, open other imports of described more than first logical reversing valves successively and extract other reagent respectively and enter described preparation pipeline, and when every kind of reagent reaches predetermined amount, stop described first peristaltic pump for some time, in a reaction warehouse, carry out a complete set of reagent of single reaction and all send into till the described preparation pipeline;
Order according to sequencing reaction, extract needed second kind of reagent, and then stop first peristaltic pump, extract then the third reagent, the 4th kind of reagent ... so circulation, in a needed reaction warehouse, carry out a complete set of reagent of single reaction and all send into described preparation pipeline, in the present embodiment, totally nine kinds of a complete set of reagent are dATP, ATP, fluorescein, APS, apyrase and the apyrase inhibitor of nine kinds of reagent: dCTP, dGTP carrying out sequencing reaction, dTTP, α position sulfo-successively.The amount of every kind of reagent, namely the opening time of first peristaltic pump sets in advance according to the requirement of sequencing reaction, it is for realize can be in a reaction warehouse carries out the cycle of once sequencing reaction that a complete set of reagent all is arranged on the described preparation pipeline, needn't stop any pump, avoid the liquid in the place ahead that termination of pumping causes to lack power, reduce flow velocity and influence sequencing reaction quality in the reaction warehouse; In order to prevent adjacent two kinds of reagent phases mutual interference, in the present embodiment, after finishing any one reagent of extraction, open described first peristaltic pump, the import of described more than first logical reversing valves is communicated with a reagent bottle that holds damping fluid (wherein having a reagent bottle to be contained with damping fluid), be used for to extract damping fluid and form fluid column, this fluid column between two kinds of adjacent reagent to isolate two kinds of adjacent reagent.
S5: open described first peristaltic pump simultaneously and be used for to extract described second peristaltic pump of damping fluid, described a complete set of reagent and damping fluid are together through entering main pipe line with described first peristaltic pump with the three-way connector that second peristaltic pump is connected;
S6 a: outlet of opening described more than second logical reversing valves, be positioned at the reagent of main pipe line and the mixed solution of damping fluid and enter corresponding reaction warehouse, the opening sequence of logical reversing valve arranges according to the reaction sequence of reaction warehouse more than second, in the present embodiment, four reaction warehouses are first, second, third, fourth reaction warehouse according to its position number consecutively.
Just realized being followed successively by the purpose that each reaction warehouse provides a complete set of reagent according to above-mentioned control method.
As priority scheme, the control method of the agent delivery assembly of present embodiment also comprises the steps:
S7: judge whether a complete set of reagent that a reaction warehouse carries out single reaction all enters this reaction warehouse, if, performing step S8;
S8: the outlet that described more than second logical reversing valves are communicated with waste liquid barrel is communicated with the import of described more than second logical reversing valves, the import of described more than first logical reversing valves be communicated with for one of them the described reagent bottle that holds damping fluid, wash described preparation pipeline and main pipe line.After a complete set of agent delivery that a reaction warehouse carries out single reaction finishes, wash described preparation pipeline and main pipe line, for a complete set of reagent of next reaction warehouse supply is prepared.
In order to prevent the interference between the adjacent reagent, in the present embodiment, also between step S3 and S4, increase following step:
S31: make the outlet of described more than first logical reversing valves and air connect for some time, this time calculates according to the time situation, select between the 0.5s-1.5s, the requirement that does not influence sequencing reaction with the adjacent reagent of the satisfied isolation of the length of formed gas column in preparing pipeline is as the criterion.
Following with reference to Figure 32 and the Controlling System that is used for the dna sequencing instrument of present embodiment is described in conjunction with Fig. 1-Figure 31.
As mentioned above, the dna sequencing instrument in the present embodiment comprises a plurality of reaction warehouses; Be used for to gather the CCD camera 110 of the optical signal that the dna sequencing reaction in each reaction warehouse produces; Be used for to support the bracing or strutting arrangement of can two dimension adjusting of CCD camera 110, bracing or strutting arrangement comprise the first servomotor (not shown) that driven CCD camera 110 switches between different reaction warehouses and driven CCD camera 110 near or away from second servomotor 326 of one of them reaction warehouse, the bracing or strutting arrangement upper edge near or be provided with a plurality of sensors 38 for detection of the CCD camera position away from the direction of reaction warehouse; And the agent delivery assembly that reaction reagent and damping fluid are provided for each reaction warehouse, the agent delivery assembly comprises that leading to reversing valves 702 and more than second for first peristaltic pump 701 that extracts sequencing reaction reagent, second peristaltic pump 711 that is used for extracting damping fluid, the more than second logical reversing valves more than 712, the first that are used for more than first logical reversing valves 702 of selective reagents and are used for the selective reaction storehouse leads to reversing valves 712 all for by motor-driven rotary valve.Namely rotated by the electric motor driving spool, one of them import of logical reversing valve 702 more than first is communicated with outlet, the reagent bottle that be connected with this import this moment just is communicated with the outlet of more than first logical reversing valves 702, when needs changed reaction reagent, the electric motor driving rotary spool rotated to desired import of opening is got final product with exporting to be communicated with.The working process of logical reversing valve 712 more than second and more than first logical reversing valves 702 are similar.When different is the motor rotation, change the position of the outlet of connecting with the import of more than second logical reversing valves 712, because each reaction warehouse is communicated with an outlet of more than second logical reversing valves 712, thereby change the reaction warehouse that is communicated with more than second logical reversing valves 712.
Shown in figure 32, the Controlling System of present embodiment comprises the PLC(programmable logic controller); PLC respectively with the controller of the controller of first servomotor and second servomotor be electrically connected to control first servomotor and second servomotor startup, stop and turning direction; PLC respectively with state startup that first peristaltic pump, second peristaltic pump be electrically connected to control first peristaltic pump and second peristaltic pump, stop and rotating speed; PLC leads to the drive-motor of reversing valves and the on position that more than second drive-motor that lead to reversing valves are electrically connected to control logical reversing valve more than first and more than second logical reversing valves with more than first respectively: PLC control more than first is led to the on position of reversing valves 702 with the reagent bottle of selection with its connection, and the on position of PLC control more than second logical reversing valves 712 is with the reaction warehouse of selection and its connection.PLC is electrically connected to control the open and close state of CCD camera with the CCD camera; PLC also connects the position signal of the CCD camera that sends with receiving sensor respectively with a plurality of sensors (sensor only is shown, and other sensors all are connected with PLC) in Figure 32.When CCD camera 110 is fitted on the reaction warehouse and reach the reaction times of expectation, open the CCD camera and obtain the optical signal that sequencing reaction produces, form image.
Shown in figure 32, the Controlling System of present embodiment also comprises the upper computer that is connected with described PLC, and described upper computer is used for arranging the sequence of control of described PLC operation, and transmits described sequence of control to described PLC.Sequence of control is the workflow of PLC, that is to say that PLC to by the control object, comprises the controller of first servomotor, controller, first peristaltic pump 701, second peristaltic pump 711, the drive-motor of more than first logical reversing valves 702, the drive-motor of more than second logical reversing valves 712 and kind and the order that CCD camera 110 transmits control signal of second servomotor.In the present embodiment, upper computer carries out serial communication by RS232 port and PLC.Upper computer is communicated by letter with the CCD camera by USB port, is used for receiving the view data that the CCD camera obtains.
Following with reference to Figure 33, and the temperature control modules of the Controlling System that is used for the dna sequencing instrument of embodiments of the invention is described in conjunction with Figure 11.
As shown in figure 33, temperature control modules comprises the rly. that is connected with PLC and the temperature sensor 561, the semiconductor heat booster 571 that are arranged in the reaction warehouse; PLC receives and is connected from the signal of temperature sensor 561 and with the primary return of rly., and the secondary loop of rly. is connected in the current supply circuit of semiconductor heat booster 571.When the temperature in the annular recesses 56 is lower than preset temperature; PLC sends connection signal to rly., the current supply circuit of semiconductor heat booster 571 is connected, when the temperature in the annular recesses 56 is higher than another preset temperature, PLC sends signal to rly. disconnects the current supply circuit of semiconductor heat booster 571, guarantees to be positioned at the temperature control of annular recesses 56 accordingly at target temperature range.In the present embodiment, rly. is electromagnetic relay.Only show the composition of the temperature control modules in the reaction warehouse among Figure 33, other reaction warehouses are identical therewith.As shown in figure 11, temperature sensor 561 and semiconductor heat booster 571 are arranged in the reaction warehouse, fixedly there is temperature sensor 561 bottom in sequencing reaction pond 51 by heat conductive silica gel, be not provided with the opposite side of annular recesses 56 at reaction tank body 5, offer the heating chamber 57 for ccontaining semiconductor heat booster 571, heating chamber 57 is opened on the side of reaction tank body 5, and fixing semiconductor heat booster 571 on its bottom surface is with to 5 heating of reaction tank body.
In this enforcement, PLC adopts the CP1H-40XDT-D-CH type of Omron, controls first servomotor and second servomotor by pulse signal respectively, and first servomotor and second servomotor are stepper-motor; PLC controls the motor speed of first peristaltic pump and second peristaltic pump by sending pulse signal.Omron CP1H-40XDT-D-CH belongs to 40 PLC of rly. output type, 8 output expansions have been introduced simultaneously again, be used for the BCD(binary coded decimal that more than first logical reversing valves and more than second lead to reversing valves) control, by carrying out the switching etc. that BCD controls to realize the on position of valve.PLC makes the current supply circuit of semiconductor heat booster control temperature by the break-make of relay.Controlling System of the present invention, make the agent delivery assembly of the dna sequencing instrument with a plurality of reaction warehouses for supplying reagent and damping fluid timely and accurately, and can make the CCD camera in time read the interior optical signal of each reaction warehouse, realized that a plurality of reactions carry out simultaneously, can check order to a plurality of samples simultaneously, improve dna sequencing efficient greatly.
Certainly, more than be preferred implementation of the present invention, should be understood that; for those skilled in the art; under the prerequisite that does not break away from the principle of the invention, can also make some improvements and modifications, these improvements and modifications also are considered as protection scope of the present invention.
Claims (4)
1. Controlling System that is used for the dna sequencing instrument, described dna sequencing instrument comprises a plurality of reaction warehouses; Be used for to gather the CCD camera of the optical signal that the dna sequencing reaction in each described reaction warehouse produces; Be used for to support the bracing or strutting arrangement of can two dimension adjusting of described CCD camera, described bracing or strutting arrangement comprise first servomotor that the driven CCD camera switches between different reaction warehouses and drive described CCD camera near or away from second servomotor of one of them reaction warehouse, described bracing or strutting arrangement upper edge near or be provided with a plurality of sensors for detection of described CCD camera position away from the direction of described reaction warehouse; And the agent delivery assembly that reaction reagent and damping fluid are provided for a plurality of described reaction warehouses, described agent delivery assembly comprise for first peristaltic pump that extracts sequencing reaction reagent, be used for extracting damping fluid second peristaltic pump, 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, described 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 described Controlling System comprises: PLC;
Described PLC respectively with the controller of the controller of described first servomotor and second servomotor be electrically connected to control described first servomotor and second servomotor startup, stop and turning direction;
Described PLC respectively with state startup that first peristaltic pump, second peristaltic pump be electrically connected to control described first peristaltic pump and second peristaltic pump, stop and rotating speed;
Described PLC is electrically connected to control the on position that logical reversing valve and more than second more than first leads to reversing valves with the drive-motor of described more than first logical reversing valves and the drive-motor of more than second logical reversing valves respectively;
Described PLC and described CCD camera are electrically connected to control the open and close state of described CCD camera;
Described PLC also is connected to receive the position signal of the CCD camera that described sensor sends respectively with described a plurality of sensors;
Described Controlling System also comprises temperature control modules, described temperature control modules comprises the rly. that is connected with described PLC and the temperature sensor and the semiconductor heat booster that are arranged in the described reaction warehouse, described PLC receives and is connected from the signal of described temperature sensor and with the primary return of described rly., and the secondary loop of described rly. is connected in the current supply circuit of described semiconductor heat booster.
2. Controlling System as claimed in claim 1 is characterized in that, also comprises the upper computer that is connected with described PLC, and described upper computer arranges the sequence of control of described PLC operation, and transmits described sequence of control to described PLC.
3. Controlling System as claimed in claim 2 is characterized in that, described upper computer is by RS232 port and described plc communication.
4. Controlling System as claimed in claim 3 is characterized in that, described upper computer is communicated by letter with described CCD camera by USB port, is used for receiving the view data that described CCD camera obtains.
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| CN105629780B (en) * | 2014-12-01 | 2018-06-01 | 深圳华大生命科学研究院 | Control device, method and the gene sequencer of gene sequencer |
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