CN102766574A - Reaction chamber for DNA sequenator - Google Patents
Reaction chamber for DNA sequenator Download PDFInfo
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- CN102766574A CN102766574A CN201210166089XA CN201210166089A CN102766574A CN 102766574 A CN102766574 A CN 102766574A CN 201210166089X A CN201210166089X A CN 201210166089XA CN 201210166089 A CN201210166089 A CN 201210166089A CN 102766574 A CN102766574 A CN 102766574A
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Abstract
The invention discloses a reaction chamber for a DNA sequenator. The reaction chamber comprises a base, a reaction tank body for sequencing reaction and a mounting base. The reaction tank body is provided with an annular groove; the mounting base is provided with a penetrating mounting cavity; a sequencing chip is mounted at one end of the mounting cavity; the mounting cavity is provided with a plurality of positioning grooves; each positioning groove is provided with a sliding tongue plate sliding into the mounting cavity to clamp the sequencing chip; a side provided with the annular groove of the reaction tank body is fixedly connected with the mounting base; the other side of the reaction tank body is affixed to the base; the reaction tank body can slide along an axial direction of a guiding rod, as well as move slightly along a radial direction of the guiding rod; and a first elastic member is arranged between the reaction tank body and the base. The present invention employs the sliding tongue plate to realize automatic clamping of the sequencing chip; the reaction tank body slides axially, and fluctuates slightly and radially on the guiding rod, so as to realize a high precision of depth of parallelism between an outer end face of an optical fiber panel and the sequencing chip, and acquisition of a sequencing signal with high signal to noise ratio.
Description
Technical field
The present invention relates to the dna sequencing technical field, be specifically related to a kind of reaction warehouse that is used for the dna sequencing appearance.
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; Through detecting the 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 up 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 resorcinolphthalein (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 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 combine to form ATP with APS, and under the catalysis of luciferase, the ATP of generation can combine to form oxyluciferin again with resorcinolphthalein, produces visible light simultaneously.Can obtain a special detected peaks through 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 accomplished, add another kind of dNTP, above-mentioned reaction is repeated, promptly 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 through fragmentation after; Application builds that storehouse reagent adds joint, microballoon, microemulsion pcr amplification, breakdown of emulsion liquid are caught, are bonded to strand; 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 sequence testing chip with micro reaction pool, sequence testing chip and sequencing reagent are mounted on the main frame, start the order-checking program through control computer according to module number and position; Sequencing reaction is carried out in robotization; Data transmission to the data analysis computingmachine that produces is accomplished the order-checking back and is used calculating analysis software and carry out work such as picture processing, sequence are read, mass analysis, sequence assembly, finally obtains 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, chemical reaction taking place on sequence testing chip, produce visible light, catch the optical signal that sequencing reaction produced through CCD (Charge Couple Device) camera, can obtain needed order-checking information.
The contriver is realizing process discovery of the present invention; Because the order-checking signal is the optical signal of extremely faint visible light wave range; Be to realize obtaining of high s/n ratio order-checking signal, require to be installed on sequence testing chip outer face and the outer face of the fibre faceplate of CCD camera front end in the reaction warehouse try one's best near, and have the high parallelism precision of trying one's best between the two; Not contaminated to guarantee the order-checking signal, can intactly be received by the CCD camera.For the ease of when sequencing reaction begins or finish, load and unload sequence testing chip fast, increase work efficiency, need a kind of loading and unloading sequence testing chip reaction warehouse easily.
Summary of the invention
To the problems referred to above; The technical problem that the present invention will solve is; A kind of reaction warehouse that is used for the dna sequencing appearance is provided, is convenient to load and unload sequence testing chip, and make the outer face of the fibre faceplate of the sequence testing chip outer face that is installed in the reaction warehouse and CCD camera front end can realize as far as possible near; And have the high parallelism precision of trying one's best, to realize obtaining of high s/n ratio order-checking signal.
For addressing the above problem, the present invention adopts following technical scheme:
A kind of reaction warehouse that is used for the dna sequencing appearance comprises base, carries out the reaction tank body of sequencing reaction and is used to install the mount pad of sequence testing chip; Offer annular recesses on the surface of said reaction tank body one side; Said mount pad is provided with the installation cavity of perforation; Said sequence testing chip is installed in the end of said installation cavity; Wherein, One end of said installation cavity is provided with the retaining ring that is used to locate said sequence testing chip; It radially is provided with the locating slot of the said inwall of a plurality of perforations the inwall upper edge of said installation cavity; A plurality of said locating slots are positioned on the axial same position of said installation cavity, and a plurality of said locating slots are along the circumference uniform distribution of said installation cavity, and being equipped with in each said locating slot slides to stretch in the said installation cavity to hold said sequence testing chip along said locating slot is close on the said retaining ring it and withdraws in the said locating slot to unclamp the slip hyoplastron of said sequence testing chip along said locating slot; The side that said reaction tank body offers annular recesses is fixedly connected with said mount pad towards said installation cavity; Said sequence testing chip and said reaction tank body offer and are provided with the gap that supplies order-checking medication flow to cross between the side of annular recesses, and said gap surrounds the sequencing reaction pond that supplies sequencing reaction through the sealing-ring that is embedded in the said groove and protrude said annular recesses; The another side of said reaction tank body is attached on the said base; And on said reaction tank body and said base, be provided with corresponding a plurality of through holes; Said reaction tank body is connected with said base through the tail rod that passes said through hole; One end of said tail rod is fixedly connected with said base, said reaction tank body through said through hole be enclosed within on the said tail rod can said tail rod axially on slide and the fine motion that can make progress in the footpath of said tail rod; Be provided with first elastomeric element between said reaction tank body and the said base.
As preferably, said reaction tank body comprises backboard and the boss that is located on the said backboard to the side protrusion of said backboard, and said annular recesses is opened on the end face of said boss; Said mount pad comprises pedestal; Said installation cavity is opened on the said pedestal, is fixedly connected between said pedestal and the said backboard to make said boss stretch in the said installation cavity and make to be provided with the said gap that the medication flow that supplies to check order is crossed between said boss and the said sequence testing chip; The side that said reaction tank body is provided with backboard is attached on the said base, and a plurality of said through hole of the corresponding setting with said base of said reaction tank body is opened on the said backboard.
As preferably; Said first elastomeric element is first spring; Said base is provided with cavity with the side that said reaction tank body is connected, and is provided with the spring lever that one or more integral body are positioned at said cavity in the said cavity, and said first spring housing is on said spring lever and stretch out said cavity; The said through hole that is located on the said reaction tank body is step-like; Said through hole is a glissile segment near a section of said base; Another section of said through hole is a construction section, and the internal diameter of said construction section is greater than said glissile segment, and said construction section and said glissile segment junction form first step; One end of said tail rod is the connection section that is connected with said base; The other end of said tail rod is provided with the boss that is used to be held in said first step place, and the middle part of said tail rod is near the sliding part of part in said glissile segment, sliding of said boss; The diameter of said glissile segment is a bit larger tham the diameter of said sliding part.
As preferably, said annular recesses is a hexagon, and when said reaction tank body was vertically placed, said hexagonal upper end was provided with liquid outlet, and said hexagonal lower end is provided with fluid inlet.
As preferably, said dna sequencing appearance comprises the bracing frame of vertical setting, and said base is rotatably connected on the support frame as described above.
As preferably, said locating slot is less near one section size of said installation cavity, is formed for the second step of spacing said slip hyoplastron; Said locating slot is provided with the baffle plate that is fixedly connected with said mount pad away from an end of said installation cavity; It is interior to hold said sequence testing chip that one end of said slip hyoplastron stretches into said installation cavity, and said slip hyoplastron is provided with and when an end of said slip hyoplastron stretches out, is against the tuck on the said second step; Be provided with between said tuck and the said baffle plate and make said slip hyoplastron stretch into second elastomeric element in the said installation cavity.
Preferred as further; Said second elastomeric element is second spring; Said second spring housing is on the said slip hyoplastron between said tuck and the said baffle plate; Said baffle plate is provided with the through hole that supplies said slip hyoplastron to pass away from an end of said installation cavity, and the inboard of said baffle plate is provided with the cover of internal diameter greater than said through hole, the end that is formed for making said second spring between said cover and the through hole against the 3rd step.
As preferably, the outer side of said mount pad is provided with the ring groove that is centered around said sequence testing chip; The front end that is used to read the CCD camera of the optical signal that occurs in the sequencing reaction on the said sequence testing chip is connected with joint flange, and the front end of said joint flange has and is used to the protruding circle that inserts said ring groove and be sealed and matched with said ring groove.
Preferred as further; Said CCD camera comprises camera body and is positioned at the intravital fibre faceplate that is used to read the optical signal that occurs in the sequencing reaction on the said sequence testing chip of said camera that the front end that one end of said fibre faceplate stretches out said camera body directly directly contacts the optical signal that sequencing reaction was produced that on the another side of sequence testing chip, carries out to obtain with a side of the sequence testing chip that is positioned at said mount pad.
As preferably; The zone that annular recesses surrounded of said reaction tank body is provided with one or more TPs; The side that said reaction tank body is connected with said base is provided with the heating chamber to said reaction tank body indent, is provided with semiconductor heat booster in the said heating chamber.
The reaction warehouse that is used for the dna sequencing appearance of the present invention; Realize the automatic chucking of sequence testing chip through the slip hyoplastron, be convenient to load and unload sequence testing chip, when the CCD camera to reaction warehouse near the time; When the outer face of the sequence testing chip in being installed on reaction warehouse contacts with the fibre faceplate outer end of CCD camera front end; Fibre faceplate drives mount pad and moves to the direction of base with the reaction tank body through promoting sequence testing chip, under the retroaction of elastomeric element, make outer face and the sequence testing chip of fibre faceplate try one's best near.And; Owing to leave the gap between the through hole of reaction tank body and the tail rod; Fibre faceplate can also drive radially unsteady a little at tail rod of mount pad and reaction tank body through promoting sequence testing chip; Make between outer face and the sequence testing chip of fibre faceplate to have the high parallelism precision of trying one's best, prevented external optical pollution, thereby reduced ground unrest, realize obtaining of high s/n ratio order-checking signal.
Description of drawings
Fig. 1 is the decomposing schematic representation of the reaction warehouse that is used for the dna sequencing appearance of embodiments of the invention;
Fig. 2 looks synoptic diagram for the master of the base of the reaction warehouse that is used for the dna sequencing appearance of embodiments of the invention;
Fig. 3 looks (broken section) synoptic diagram for the left side of Fig. 2;
Fig. 4 is the structural representation that the reaction tank body of the reaction warehouse that is used for the dna sequencing appearance of embodiments of the invention cooperates with tail rod;
Fig. 5 looks (broken section) synoptic diagram for the left side of Fig. 4;
Fig. 6 is the structural representation of the reaction tank body among Fig. 5;
Fig. 7 is the structural representation of the tail rod among Fig. 5;
Fig. 8 is that the A-A of Fig. 4 is to sectional drawing;
Fig. 9 looks synoptic diagram for the master of the mount pad of the reaction warehouse that is used for the dna sequencing appearance of embodiments of the invention;
Figure 10 looks (broken section) synoptic diagram for the left side of Fig. 9;
Figure 11 is that the B-B of Fig. 9 is to sectional schematic diagram;
Figure 12 is that the C-C of Figure 11 is to synoptic diagram;
Figure 13 is the front view of the slip hyoplastron among Figure 11;
Figure 14 is the left view of Figure 13;
Figure 15 is the perspective view of the baffle plate among Figure 11;
Figure 16 is the side-view of baffle plate shown in Figure 15;
Figure 17 is that the D-D of Figure 16 is to sectional view;
Figure 18 is the structural representation (front end of camera body is connected with joint flange) of the CCD camera that is complementary with the reaction warehouse that is used for the dna sequencing appearance of embodiments of the invention;
Figure 19 is the front view of the joint flange among Figure 18;
Figure 20 is that the E-E of Figure 19 is to sectional view;
Structural representation (broken section) when Figure 21 is connected with the CCD camera for the reaction warehouse that is used for the dna sequencing appearance of embodiments of the invention.
Main description of reference numerals
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-boss
510-fluid inlet 511-liquid outlet 561-TP
571-semiconductor heat booster 572-shut
605-glissile segment 606-construction section 610-first step
612-screw 621-tuck 623-second step
The 633-second spring 841-screw
Embodiment
Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail, but not as to qualification of the present invention.
As shown in Figure 1, the reaction warehouse that is used for the dna sequencing appearance of embodiments of the invention comprises base 8, carries out the reaction tank body 5 of sequencing reaction and is used to install 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 fixedly connected with base 8; Reaction tank body 5 is enclosed within on the tail rod 9, tail rod 9 axially on slide and the fine motion that can make progress in the footpath 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.Specify below in conjunction with accompanying drawing 1-21 present embodiment the reaction warehouse that is used for the dna sequencing appearance detailed structure with and working process when being connected with the CCD camera.
Like Fig. 1, Fig. 2 and shown in Figure 3, 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.Each spring lever 85 is attend sheathed one first spring, 81, the first springs 81 of branch and is stretched out cavity 80.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.
Like Fig. 1, Fig. 4 and shown in Figure 8; Reaction tank body 5 comprises backboard 54 and is located on the said backboard 54 and the boss 55 of the side protrusion of toward back plate 54; The end face of said boss 55 is provided with the annular recesses 56 of sealing, and the zone that the annular recesses 56 of sealing is surrounded is sequencing reaction pond 51; As shown in Figure 4, be embedded with sealing-ring 58 in the annular recesses 56, the end face of sealing-ring 58 protrusion boss 55, to surround sequencing reaction pond 51, wherein the end face of boss 55 is smooth as much as possible.Like Fig. 1, Fig. 4, Fig. 5 and shown in Figure 6; The side that reaction tank body 5 is provided with backboard 54 is attached on the base 8, and the boss 55 of reaction tank body 5 is provided with four first through holes, and said first through hole is step-like; One section near base is glissile segment 605; Another section of first through hole is a construction section 606, and the internal diameter of construction section 606 forms first step 610 greater than glissile segment 605.Like Fig. 4-shown in Figure 7; 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 with the boss 93 that is used to be held in first step 610 places, and the middle part of tail rod 9 is to be used for the sliding part 92 that cooperates with glissile segment 605 near the part of boss 93.As shown in Figure 9, connection section 91 is provided with screw thread, and is as shown in Figure 4, and the end of the boss 93 of tail rod 9 is provided with the cutter groove that is used to cooperate screwdriver.As shown in Figure 1, tail rod 9 passes first through hole and second through hole 82 on the base 8 on the reaction tank body 5, and the back that makes connection section 91 expose base 8 is fastening through nut then, thereby realizes being fixedly 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 somewhat 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 boss 93; So that reaction tank body 5 is being slidingly connected at 9 last times of tail rod; Through boss 93 position-limiting action with first step 610, tighten with connection section 91 on during the nut of thread fit, 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 to have certain preload, it is exactly in order to be attached to 8 last times of base when reaction tank body 5 that above-mentioned first spring 81 stretches out cavity 80, and first spring 81 can be given reaction tank body 5 certain function power.
Like Fig. 1, Fig. 9, Figure 10 and shown in Figure 11; The mount pad 6 of sequence testing chip comprises pedestal 63 and is opened in the installation cavity 69 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; Being fixedly 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, is provided with the gap that supplies order-checking medication flow to cross between the end face of sequence testing chip 7 and boss 55, and this gap is through being embedded at sealing-ring 58 sealings in the annular recesses 56; The size in said gap is about 0.2mm, and sealing-ring 58 convex hole connected in stars 56 contact the sequencing reaction pond 51 with the confession sequencing reaction that surrounds sealing with sequence testing chip 7.
Like Fig. 4, shown in Figure 8; In the present embodiment, this annular recesses 56 is a hexagon, and 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 flow hydrokinetics advantageous shape that balanced flow is crossed in reaction warehouse.As shown in Figure 8; Bottom in sequencing reaction pond 51 is fixed with TP 561 through heat conductive silica gel, is not provided with the opposite side of annular recesses 56 at reaction tank body 5, offers the heating chamber 57 that is used 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 is controlled at 35 ± 1 ℃.In the present embodiment, heating chamber 57 is through shut 572 sealings, so that be adjacent on the mount pad 6 of sequence testing chip with reaction tank body 5.
Like Fig. 1, Fig. 9 and shown in Figure 10; The pedestal 63 of the mount pad 6 of sequence testing chip is provided with four third through-holes; Four set screws 612 are connected on the backboard 54 of four screws 61 of the mount pad 6 of sequence testing chip through passing these four third through-holes and reaction tank body 5, thereby realize being fixedly connected between the backboard 54 of pedestal 63 and reaction tank body 5 of mount pad 6 of sequence testing chip.Like Fig. 9-shown in Figure 12; The pedestal 63 of the mount pad 6 of sequence testing chip is provided with and comprises the installation cavity 69 that is used for ccontaining sequence testing chip 7; One end of installation cavity 69 is provided with the retaining ring 67 that is used to locate 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; Its quantity can be even numbers such as two or four; In order stable holding to live sequence testing chip 7, the circumferential direction uniform distribution of a plurality of locating slots on the axial same position of installation cavity 69.Certainly, more a plurality of locating slots can also be set, but more locating slot can cause the inconvenience of operation in order to improve holding force.In the present embodiment, locating slot is two, and for easy to process, locating slot is a circle hole shape.As shown in Figure 2, locating slot is less near one section size of installation cavity 69, is formed for the second step 623 of spacing slip hyoplastron 62; Be locating slot near a section of installation cavity 69 be a less circular hole, rest part is a bigger circular hole.Like Fig. 6 and shown in Figure 7, slip hyoplastron 62 is provided with the tuck 621 of protrusion.As shown in Figure 4, when slip hyoplastron 62 when installation cavity 69 stretches into, the tuck 621 of slip hyoplastron 62 is resisted against said second step 623 places, 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 that is fixedly connected with mount pad 6; Between the tuck 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 tuck 621 and the baffle plate 64.Like Figure 10, shown in Figure 11, on the surface of the mount pad 6 of sequence testing chip, have ring groove 66, 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 is surrounded.
As shown in Figure 5, baffle plate 64 is fixedly connected with mount pad 6 through two screws 41, is convenient to installation and removal.Second spring 633 is enclosed within on the slip hyoplastron 62 between tuck 621 and the baffle plate 64; Like Fig. 8-shown in Figure 10; Baffle plate 64 is provided with the through hole 44 that supplies slip hyoplastron 62 to pass away from an end of installation cavity 69; The inboard of baffle plate 64 is provided with the cover 45 of internal diameter greater than 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 that extend into of slip hyoplastron 62, make in its withdrawal locating slot; Tuck 621 extruding second spring 633 of slip hyoplastron 62; Unclamp slip hyoplastron 62 after putting into sequence testing chip 7 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,, shown in figure 21 as preferably; Dna sequencing appearance in the present embodiment comprises bracing frame 84, and base 8 is rotatably connected on the bracing frame 84 through rotating shaft as shown in figures 1 and 3 87, when installing or unloading sequence testing chip 7; Let base 8 be positioned at level attitude; When carrying out sequencing reaction, base 8 is positioned at vertical position shown in figure 21, and base 8 is fixedly connected with bracing frame 84 through screw 841; One end band knob of screw 841, the other end is provided with connecting thread.
Shown in figure 18; Be used for comprising camera body 1 and the fibre faceplate 4 that is used to read the optical signal that occurs in the sequencing reaction on the said sequence testing chip 7 that is positioned at said camera body 1 that the side that an end of fibre faceplate 4 stretches out the sequence testing chip 7 of the direct mount pad 6 (structure of the mount pad 6 of sequence testing chip sees above and is described in detail) with the sequence testing chip that is positioned at the dna sequencing appearance of the front end of said camera body 1 directly contacts to obtain the optical signal that sequencing reaction was produced on the opposite side that occurs in sequence testing chip 7 with the CCD camera of the reaction warehouse coupling that is used for the dna sequencing appearance of present embodiment.In the present embodiment, as preferably, the optical fiber core diameter of said fibre faceplate is 6 μ m, and the optical fiber core diameter can be selected between 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.
Like Figure 18, shown in Figure 21, in the present embodiment, as preferably, the front end of camera body 1 is connected with joint flange 2.Like Figure 18, Figure 19 and shown in Figure 20, the front end of camera body 1 have the ring groove 66 that is used to insert on the mount pad 6 of being located at sequence testing chip and with ring groove 66 close-fitting protruding circles 21.Joint flange 2 and camera body 1 are connected through bolt, and the fibre faceplate 4 in the camera body 1 is positioned at the zone that protruding circle 21 is surrounded, and promptly 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 said 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 through be provided with can with the direct-coupled fibre faceplate 4 of sequence testing chip; The faint visible light that sequencing reaction produced that on sequence testing chip 7, carries out 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.Shown in figure 21; When the CCD camera during near the mount pad 6 of sequence testing chip; The joint flange 2 that is installed in camera body 1 front end is near the sequence testing chip 7 on the mount pad 6 of sequence testing chip, and in the ring groove 66 on the mount pad 6 of the protruding circle 21 entering sequence testing chips of the ring-type on the joint flange 2, 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, the luminous energy that further guarantees sequencing reaction and produced is received by fibre faceplate 4, guarantees obtaining of order-checking signal.Through have the joint flange 2 of protruding circle 21 in camera body 1 front end increase; And on the mount pad 6 of sequence testing chip, offer corresponding annular groove 66; 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 is used for the dna sequencing appearance that Figure 21 shows present embodiment is connected with the CCD camera promptly is used for the reaction warehouse of the dna sequencing appearance structural representation when in running order.Shown in figure 21; When carrying out sequencing reaction; The camera body 1 of CCD camera is near reaction warehouse, and 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 fixed connection 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 receives the reactive force of first spring 81 between reaction tank body 5 and the base 8, is close to each other 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 receive 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 receiving the reactive force of first spring 81, and 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 through in the adjustment of tail rod 9 on axially; Realized pressing close to as much as possible between the outer face of outer face and fibre faceplate 4 of sequence testing chip 7; 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 outer face and the sequence testing chip 7 of fibre faceplate 4, guaranteed to greatest extent at fibre faceplate 4 to have prevented external optical pollution when sequence testing chip 7 reads the fluorescent signal of sequencing reaction; Thereby reduced ground unrest, realized obtaining of high s/n ratio order-checking signal.
Certainly, the above is a 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 improvement and retouching, these improvement and retouching also are regarded as protection scope of the present invention.
Claims (10)
1. a reaction warehouse that is used for the dna sequencing appearance is characterized in that, comprises base, carries out the reaction tank body of sequencing reaction and is used to install the mount pad of sequence testing chip;
Offer annular recesses on the surface of said reaction tank body one side;
Said mount pad is provided with the installation cavity of perforation; Said sequence testing chip is installed in the end of said installation cavity; Wherein, One end of said installation cavity is provided with the retaining ring that is used to locate said sequence testing chip; It radially is provided with the locating slot of the said inwall of a plurality of perforations the inwall upper edge of said installation cavity; A plurality of said locating slots are positioned on the axial same position of said installation cavity, and a plurality of said locating slots are along the circumference uniform distribution of said installation cavity, and being equipped with in each said locating slot slides to stretch in the said installation cavity to hold said sequence testing chip along said locating slot is close on the said retaining ring it and withdraws in the said locating slot to unclamp the slip hyoplastron of said sequence testing chip along said locating slot;
The side that said reaction tank body offers annular recesses is fixedly connected with said mount pad towards said installation cavity; Said sequence testing chip and said reaction tank body offer and are provided with the gap that supplies order-checking medication flow to cross between the side of annular recesses, and said gap surrounds the sequencing reaction pond that supplies sequencing reaction through the sealing-ring that is embedded in the said groove and protrude said annular recesses;
The another side of said reaction tank body is attached on the said base; And on said reaction tank body and said base, be provided with corresponding a plurality of through holes; Said reaction tank body is connected with said base through the tail rod that passes said through hole; One end of said tail rod is fixedly connected with said base, said reaction tank body through said through hole be enclosed within on the said tail rod can said tail rod axially on slide and the fine motion that can make progress in the footpath of said tail rod;
Be provided with first elastomeric element between said reaction tank body and the said base.
2. the reaction warehouse that is used for the dna sequencing appearance as claimed in claim 1 is characterized in that, said reaction tank body comprises backboard and be located at the boss that protrudes to a side of said backboard on the said backboard that said annular recesses is opened on the end face of said boss;
Said mount pad comprises pedestal; Said installation cavity is opened on the said pedestal, is fixedly connected between said pedestal and the said backboard to make said boss stretch in the said installation cavity and make to be provided with the said gap that the medication flow that supplies to check order is crossed between said boss and the said sequence testing chip;
The side that said reaction tank body is provided with backboard is attached on the said base, and a plurality of said through hole of the corresponding setting with said base of said reaction tank body is opened on the said backboard.
3. according to claim 1 or claim 2 the reaction warehouse that is used for the dna sequencing appearance; It is characterized in that; Said first elastomeric element is first spring; Said base is provided with cavity with the side that said reaction tank body is connected, and is provided with the spring lever that one or more integral body are positioned at said cavity in the said cavity, and said first spring housing is on said spring lever and stretch out said cavity;
The said through hole that is located on the said reaction tank body is step-like; Said through hole is a glissile segment near a section of said base; Another section of said through hole is a construction section, and the internal diameter of said construction section is greater than said glissile segment, and said construction section and said glissile segment junction form first step;
One end of said tail rod is the connection section that is connected with said base; The other end of said tail rod is provided with the boss that is used to be held in said first step place, and the middle part of said tail rod is near the sliding part of part in said glissile segment, sliding of said boss;
The diameter of said glissile segment is a bit larger tham the diameter of said sliding part.
4. according to claim 1 or claim 2 the reaction warehouse that is used for the dna sequencing appearance is characterized in that said annular recesses is a hexagon, and when said reaction tank body was vertically placed, said hexagonal upper end was provided with liquid outlet, and said hexagonal lower end is provided with fluid inlet.
5. according to claim 1 or claim 2 the reaction warehouse that is used for the dna sequencing appearance is characterized in that said dna sequencing appearance comprises the bracing frame of vertical setting, and said base is rotatably connected on the support frame as described above.
6. according to claim 1 or claim 2 the reaction warehouse that is used for the dna sequencing appearance is characterized in that said locating slot is less near one section size of said installation cavity, is formed for the second step of spacing said slip hyoplastron; Said locating slot is provided with the baffle plate that is fixedly connected with said mount pad away from an end of said installation cavity;
It is interior to hold said sequence testing chip that one end of said slip hyoplastron stretches into said installation cavity, and said slip hyoplastron is provided with and when an end of said slip hyoplastron stretches out, is against the tuck on the said second step;
Be provided with between said tuck and the said baffle plate and make said slip hyoplastron stretch into second elastomeric element in the said installation cavity.
7. the reaction warehouse that is used for the dna sequencing appearance as claimed in claim 6; It is characterized in that; Said second elastomeric element is second spring, and said second spring housing is on the said slip hyoplastron between said tuck and the said baffle plate, and said baffle plate is provided with the through hole that supplies said slip hyoplastron to pass away from an end of said installation cavity; The inboard of said baffle plate is provided with the cover of internal diameter greater than said through hole, the end that is formed for making said second spring between said cover and the through hole against the 3rd step.
8. according to claim 1 or claim 2 the reaction warehouse that is used for the dna sequencing appearance is characterized in that the outer side of said mount pad is provided with the ring groove that is centered around said sequence testing chip; The front end that is used to read the CCD camera of the optical signal that occurs in the sequencing reaction on the said sequence testing chip is connected with joint flange, and the front end of said joint flange has and is used to the protruding circle that inserts said ring groove and be sealed and matched with said ring groove.
9. the reaction warehouse that is used for the dna sequencing appearance as claimed in claim 8; It is characterized in that; Said CCD camera comprises camera body and is positioned at the intravital fibre faceplate that is used to read the optical signal that occurs in the sequencing reaction on the said sequence testing chip of said camera that the front end that one end of said fibre faceplate stretches out said camera body directly directly contacts the optical signal that sequencing reaction was produced that on the another side of sequence testing chip, carries out to obtain with a side of the sequence testing chip that is positioned at said mount pad.
10. according to claim 1 or claim 2 the reaction warehouse that is used for the dna sequencing appearance; It is characterized in that; The zone that annular recesses surrounded of said reaction tank body is provided with one or more TPs; The side that said reaction tank body is connected with said base is provided with the heating chamber to said reaction tank body indent, is provided with semiconductor heat booster in the said heating chamber.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210166089XA CN102766574B (en) | 2012-05-24 | 2012-05-24 | Reaction chamber for DNA sequenator |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210166089XA CN102766574B (en) | 2012-05-24 | 2012-05-24 | Reaction chamber for DNA sequenator |
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| CN102766574B CN102766574B (en) | 2013-12-25 |
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| CN201210166089XA Active CN102766574B (en) | 2012-05-24 | 2012-05-24 | Reaction chamber for DNA sequenator |
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Owner name: JILIN ZHONGKE ZIXIN TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: SEMICONDUCTOR INST., CHINESE ACADEMY OF SCIENCES Effective date: 20131223 |
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