CN102766574B - Reaction chamber for DNA sequenator - Google Patents
Reaction chamber for DNA sequenator Download PDFInfo
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- CN102766574B CN102766574B CN201210166089XA CN201210166089A CN102766574B CN 102766574 B CN102766574 B CN 102766574B CN 201210166089X A CN201210166089X A CN 201210166089XA CN 201210166089 A CN201210166089 A CN 201210166089A CN 102766574 B CN102766574 B CN 102766574B
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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 for DNA sequencer.
Background technology
In the DNA sequencing technical field, tetra-sodium sequencing technologies (pyrosequencing), be a kind of novel enzyme cascade sequencing technologies 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 the enzyme cascade chemiluminescence reaction in 4 kinds of enzymatic same reaction systems.The principle of tetra-sodium sequencing technologies is: after primer and 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 release coupling of the polymerization of each dNTP on primer and an optical signal is 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 consists of reaction substrate, strand to be measured, sequencing primer and four kinds of enzymes.Reaction substrate is 5 '-phosphinylidyne sulfuric acid (adenosine-5 '-phosphosulfate, APS) and fluorescein (luciferin).
At each, take turns in sequencing reaction, only add a kind of deoxynucleotide triphosphoric acid (dNTP) in reaction system, if it can just with the next base pairing of DNA profiling, 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 be with APS in conjunction with forming ATP, and under the catalysis of luciferase, the ATP of generation can, with fluorescein in conjunction with forming oxyluciferin, produce visible ray again simultaneously.Can obtain a special detected peaks by Weak light detection device and process software, the height of peak value is directly proportional with the base number be complementary.If the dNTP added can not with the next base pairing of DNA profiling, above-mentionedly instead would not occur, just there is no detected peaks yet.In reaction system, remaining dNTP and residual a small amount of ATP degrade under the effect of Apyrase.After last round of reaction completes, add another kind of dNTP, above-mentioned reaction is repeated, can read DNA sequence dna information accurately according to the peak value figure obtained.
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 microballoon, the application add model by 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 main frame, start the order-checking program by controlling computer according to module number and position, sequencing reaction is carried out in automatization, the data transmission produced is to the data analysis computer, after completing order-checking, the computation analysis software carries out the image processing, sequence is read, mass analysis, the 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 approximately 3,000,000 micro reaction pools on 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 occurring on sequence testing chip, produces visible ray, by CCD(Charge Couple Device) camera catches the optical signal that sequencing reaction produces, and can obtain needed order-checking information.
The contriver is realizing process discovery of the present invention, due to the order-checking signal optical signal that is extremely faint visible light wave range, for realizing obtaining of high s/n ratio order-checking signal, it is as far as possible close that requirement is installed on the outer face of fibre faceplate of sequence testing chip outer face in reaction warehouse and CCD camera front end, and there is 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 starts or finish, load and unload fast sequence testing chip, increase work efficiency, need a kind of handling sequence testing chip reaction warehouse easily.
Summary of the invention
For the problems referred to above, the technical problem to be solved in the present invention is, a kind of reaction warehouse for DNA sequencer is provided, be convenient to load and unload sequence testing chip, and the outer face that makes to be installed on the fibre faceplate of sequence testing chip outer face in reaction warehouse and CCD camera front end can be realized as far as possible close, and there is 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 for DNA sequencer, comprise base, carry out the reaction tank body of sequencing reaction and for the mount pad of sequence testing chip is installed, offer annular recesses on the surface of described reaction tank body one side, described mount pad is provided with the installation cavity of perforation, described sequence testing chip is arranged in an end of described installation cavity, wherein, one end of described installation cavity is provided with for locating the retaining ring of described sequence testing chip, radially be provided with the locating slot of the described inwall of a plurality of perforations on the inwall of described installation cavity along it, a plurality of described locating slots are positioned on the axial same position of described installation cavity, a plurality of described locating slots distribute along the even circumferential of described installation cavity, in each described locating slot, being equipped with slides to stretch in described installation cavity to hold described sequence testing chip along described locating slot is close on described retaining ring it and retracts in described locating slot to unclamp the slip hyoplastron of described sequence testing chip along described locating slot, the side that described reaction tank body offers annular recesses is fixedly connected with described mount pad towards described installation cavity, described sequence testing chip and described reaction tank body offer between the side of annular recesses and are provided with the gap of flowing through for the order-checking liquid, and described gap surrounds the sequencing reaction pond for sequencing reaction by the sealing-ring that is embedded in described groove and protrude described annular recesses, the another side of described reaction tank body is attached on described base, and be provided with corresponding a plurality of through holes on described reaction tank body and described base, described reaction tank body is connected with described base by the tail rod through described through hole, one end of described tail rod is fixedly connected with described base, described reaction tank body by described through hole be enclosed within on described tail rod can described tail rod axially on slide and the fine motion that can make progress in the footpath of described tail rod, be provided with the first elastomeric element between described reaction tank body and described base.
As preferably, described reaction tank body comprises backboard and is located at the boss protruded to a side of described backboard on described backboard, and described annular recesses is opened on the end face of described boss; Described mount pad comprises pedestal, described installation cavity is opened on described pedestal, is fixedly connected with between described pedestal and described backboard and makes described boss stretch in described installation cavity and make between described boss and described sequence testing chip to be provided with the described gap of flowing through for the liquid that checks order; The side that described reaction tank body is provided with backboard is attached on described base, and a plurality of described through hole of described reaction tank body setting corresponding to described base is opened on described backboard.
As preferably, described the first elastomeric element is the first spring, described base is provided with cavity with the side that described reaction tank body is connected, and is provided with the spring lever that one or more integral body is positioned at described cavity in described cavity, and described the first spring housing is on described spring lever and stretch out described cavity; The described through hole be located on described reaction tank body is step-like, one section of the close described base of described through hole is glissile segment, another section of described through hole is construction section, and the internal diameter of described construction section is greater than described glissile segment, and described construction section and described glissile segment junction form first step; One end of described tail rod is the connection section be connected with described base, the other end of described tail rod is provided with for being held in the lug boss at described first step place, and the middle part of described tail rod is near the sliding part of part for sliding in described glissile segment of described lug boss; The diameter of described glissile segment is a bit larger tham the diameter of described sliding part.
As preferably, described annular recesses is hexagon, and when described reaction tank body is vertically placed, described hexagonal upper end is provided with liquid outlet, and described hexagonal lower end is provided with fluid inlet.
As preferably, described DNA sequencer comprises the bracing frame of vertical setting, and described base is rotatably connected on support frame as described above.
As preferably, described locating slot is less near the size of a section of described installation cavity, is formed for the second step of spacing described slip hyoplastron; Described locating slot is provided with away from an end of described installation cavity the baffle plate be fixedly connected with described mount pad; It is interior to hold described sequence testing chip that one end of described slip hyoplastron stretches into described installation cavity, and described slip hyoplastron is provided with and is against the protuberance on described second step when an end of described slip hyoplastron stretches out; Be provided with between described protuberance and described baffle plate and make described slip hyoplastron stretch into the second elastomeric element in described installation cavity.
As preferred further, described the second elastomeric element is the second spring, described the second spring housing is on the described slip hyoplastron between described protuberance and described baffle plate, described baffle plate is provided with the through hole passed away from an end of described installation cavity for described slip hyoplastron, the inboard of described baffle plate is provided with the cover that internal diameter is greater than described through hole, the end that is formed for making described the second spring between described cover and through hole against the 3rd step.
As preferably, the outer side of described mount pad is provided with the ring groove that is centered around described sequence testing chip; Be connected with joint flange for the front end of CCD camera that reads the optical signal that occurs in the sequencing reaction on described sequence testing chip, the front end of described joint flange has the flange collar for inserting described ring groove and being sealed and matched with described ring groove.
As preferred further, described CCD camera comprise camera body and be positioned at described camera body for reading the fibre faceplate of the optical signal that occurs in the sequencing reaction on described sequence testing chip, an end of described fibre faceplate stretches out that the front end of described camera body is direct directly contacts to obtain with a side of the sequence testing chip that is positioned at described mount pad the optical signal that the sequencing reaction that carries out on the another side of sequence testing chip produces.
As preferably, the zone that the annular recesses of described reaction tank body surrounds is provided with one or more temperature sensors, the side that described reaction tank body is connected with described base is provided with to the heating chamber of described reaction tank body indent, in described heating chamber, is provided with semiconductor heat booster.
Reaction warehouse for DNA sequencer of the present invention, realize the automatic clamping of sequence testing chip by the slip hyoplastron, be convenient to load and unload sequence testing chip, when the CCD camera is close to reaction warehouse, 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 reaction tank body and moves to the direction of base by promoting sequence testing chip, under the retroaction of elastomeric element, makes the outer face of fibre faceplate and sequence testing chip as far as possible close.And, owing between the through hole of reaction tank body and tail rod, leaving gap, fibre faceplate can also drive radially unsteady a little at tail rod of mount pad and reaction tank body by promoting sequence testing chip, make to there is the high parallelism precision of trying one's best between the outer face of fibre faceplate and sequence testing chip, prevented external optical pollution, thereby reduced ground unrest, realized obtaining of high s/n ratio order-checking signal.
The accompanying drawing explanation
The decomposing schematic representation of the reaction warehouse for DNA sequencer that Fig. 1 is embodiments of the invention;
The master of the base of the reaction warehouse for DNA sequencer that Fig. 2 is embodiments of the invention looks schematic diagram;
(broken section) schematic diagram is looked in the left side that Fig. 3 is Fig. 2;
The structural representation that the reaction tank body of the reaction warehouse for DNA sequencer that Fig. 4 is embodiments of the invention coordinates with tail rod;
(broken section) schematic diagram is looked in the left side that Fig. 5 is Fig. 4;
The structural representation that Fig. 6 is the reaction tank body in Fig. 5;
The structural representation that Fig. 7 is the tail rod in Fig. 5;
The A-A that Fig. 8 is Fig. 4 is to sectional drawing;
The master of the mount pad of the reaction warehouse for DNA sequencer that Fig. 9 is embodiments of the invention looks schematic diagram;
(broken section) schematic diagram is looked in the left side that Figure 10 is Fig. 9;
The B-B that Figure 11 is Fig. 9 is to sectional schematic diagram;
The C-C that Figure 12 is Figure 11 is to schematic diagram;
The front view that Figure 13 is the slip hyoplastron in Figure 11;
The left view that Figure 14 is Figure 13;
The perspective view that Figure 15 is the baffle plate in Figure 11;
The side-view that Figure 16 is the baffle plate shown in Figure 15;
The D-D that Figure 17 is Figure 16 is to sectional view;
The structural representation (front end of camera body is connected with joint flange) that Figure 18 is the CCD camera that is complementary with the reaction warehouse for DNA sequencer of embodiments of the invention;
The front view that Figure 19 is the joint flange in Figure 18;
The E-E that Figure 20 is Figure 19 is to sectional view;
Structural representation (broken section) when the reaction warehouse for DNA sequencer that Figure 21 is embodiments of the invention is connected with the CCD camera.
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
8-base 9-tail rod 21-flange collar
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
81-first spring 82-the second through hole 84-bracing frame
85-spring lever 86-spring support 87-rotating shaft
91-connection section 92-sliding part 93-lug boss
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-the second spring 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.
As shown in Figure 1, the reaction warehouse for DNA sequencer of embodiments of the invention, comprise base 8, carry out the reaction tank body 5 of sequencing reaction and for the mount pad 6 of sequence testing chip that sequence testing chip 7 is installed; The mount pad 6 of sequence testing chip is fixedly connected on reaction tank body 5, base 8 and reaction tank body 5 are provided with through 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 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 the first elastomeric element between reaction tank body 5 and base 8, as shown in Figure 1, in the present embodiment, the first elastomeric element is preferably the first spring 81, spring has simple in structure, is convenient to the advantage of installing, and certainly can also select other elastomeric elements that can realize said function to realize.Below in conjunction with accompanying drawing 1-21, describe in detail the present embodiment the reaction warehouse for DNA sequencer detailed construction with and working process while being connected with the CCD camera.
As shown in Figure 1, Figure 2 and Figure 3, a side of base 8 is provided with cavity 80, and cavity 80 is interior is provided with two spring levers 85 side by side, and spring lever 85 integral body are positioned at cavity 80, can not stretch out cavity 80.On each spring lever 85, sheathed first spring 81, the first springs 81 of branch stretch 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 spring support 86, and the spring support 86 of increase can reduce the required length of the first spring 81.It is for the ease of miscellaneous part is installed that cavity 80 is set.
As shown in Fig. 1, Fig. 4 and Fig. 8, reaction tank body 5 comprises backboard 54 and is located at the boss 55 that on described backboard 54, also 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 4, be embedded with sealing-ring 58 in 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 shown in Fig. 1, Fig. 4, Fig. 5 and Fig. 6, the side that reaction tank body 5 is provided with backboard 54 is attached on base 8, the boss 55 of reaction tank body 5 is provided with four the first through holes, described the first through hole is step-like, one section near base is glissile segment 605, another section of the first through hole is construction section 606, and the internal diameter of construction section 606 is greater than glissile segment 605, forms first step 610.As Figure 4-Figure 7, one end of tail rod 9 is the connection section 91 be connected with base 8, the other end of tail rod 9 is provided with for being held in the lug boss 93 at first step 610 places, and the middle part of tail rod 9 is the sliding part 92 for coordinating with glissile segment 605 near the part of lug boss 93.Connection section 91 is provided with screw thread, and as shown in Figure 4, the end of the lug boss 93 of tail rod 9 is provided with for coordinating the cutter groove of screwdriver.As shown in Figure 1, tail rod 9 is through the first through hole on reaction tank body 5 and the second through hole 82 on base 8, and being fixedly connected with between tail rod 9 and base 8 then by fastening nuts, thereby realized in the back that makes connection section 91 expose base 8.In order to make to realize being slidably connected between reaction tank body 5 and 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 realize tail rod 9 axially on slip, the fine motion again can realization response pond body 5 made 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 slidably connected on tail rod 9 by reaction tank body 5, position-limiting action by 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 the first spring 81, make to there is certain preload between reaction tank body 5 and base 8, it is exactly in order to be attached on base 8 when reaction tank body 5 that above-mentioned the first spring 81 stretches out cavity 80, the first spring 81 can be given the certain reactive force of reaction tank body 5.
As Fig. 1, Fig. 9, shown in Figure 10 and Figure 11, the mount pad 6 of sequence testing chip comprises pedestal 63 and is opened in the installation cavity 69 on 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, be fixedly connected with boss 55 is stretched in installation cavity 69 between pedestal 63 and backboard 54, sequence testing chip 7 is fixed in the outer end of installation cavity 69, be provided with the gap of flowing through for the order-checking liquid between the end face of sequence testing chip 7 and boss 55, this gap is by being embedded at sealing-ring 58 sealings in annular recesses 56, the size in described gap is about 0.2mm, 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 shown in Fig. 4, Fig. 8, 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, in hexagonal upper end, forms the liquid stream 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 temperature sensor 561 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 accommodating 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, guarantee that the temperature that is positioned at annular recesses 56 is controlled 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 shown in Fig. 1, Fig. 9 and Figure 10, the pedestal 63 of the mount pad 6 of sequence testing chip is provided with four third through-holes, on the backboard 54 of the mount pad 6 of sequence testing chip by four screws 61 through these four third through-holes and reaction tank body 5, four set screws 612 are connected, thereby realize being fixedly connected with between the backboard 54 of pedestal 63 and reaction tank body 5 of mount pad 6 of sequence testing chip.As shown in Fig. 9-Figure 12, the pedestal 63 of the mount pad 6 of sequence testing chip is provided with the installation cavity 69 comprised for accommodating sequence testing chip 7, one end of installation cavity 69 is provided with the retaining ring 67 for locating sequence testing chip 7, radially be provided with locating slot along it on the inwall of installation cavity 69, in locating slot, be provided with along locating slot slide stretch in installation cavity 69 with hold sequence testing chip 7 make it be attached on retaining ring 67 or in the retraction locating slot to unclamp the slip hyoplastron 62 of sequence testing chip 7.Distance when slip hyoplastron 62 stretches out and between retaining ring 67 equals the thickness of sequence testing chip just, when slip hyoplastron 62 stretches out, can make sequence testing chip 7 be 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 the even numbers such as two or four, in order stable holding to live sequence testing chip 7, a plurality of locating slots are uniformly distributed 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 12, locating slot is less near the size of a section 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 larger circular hole.As shown in Figure 13 and Figure 14, slip hyoplastron 62 is provided with the protuberance 621 of protrusion.As shown in figure 12, when slip hyoplastron 62 stretches into to installation cavity 69, the protuberance 621 of slip hyoplastron 62 is resisted against described second step 623 places, and an end of slip hyoplastron 62 stretches in installation cavity 69 to hold sequence testing chip 7.The outer end of locating slot is provided with the baffle plate 64 be fixedly connected with mount pad 6, be provided with the second spring 633 between the protuberance 621 of slip hyoplastron 62 and baffle plate 64, in the present embodiment, the second spring 633 is enclosed within the other end of slip hyoplastron 62, and the two ends of the second spring 633 are resisted against respectively on protuberance 621 and baffle plate 64.As shown in Figure 10, 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 surrounds.
As shown in figure 12, baffle plate 64 is fixedly connected with mount pad 6 by two screws 41, is convenient to installation and removal.The second spring 633 is enclosed within on the slip hyoplastron 62 between protuberance 621 and baffle plate 64, as shown in Figure 15-Figure 17, baffle plate 64 is provided with the through hole 44 passed away from an end of installation cavity 69 for slip hyoplastron 62, the inboard of baffle plate 64 is provided with the cover 45 that internal diameter is greater than through hole 44, overlap 45 and through hole 44 between be formed for making the second spring 633 end against the 3rd step 49.In the present embodiment, the 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 the second spring 633.When sequence testing chip 7 is installed, stir the end in installation cavity 69 that extend into of slip hyoplastron 62, make in its retraction locating slot, protuberance 621 extruding second springs 633 of slip hyoplastron 62, unclamp slip hyoplastron 62 after sequence testing chip 7 is put into to installation cavity 69 and is attached on retaining ring 67, slip hyoplastron 62 stretches out under the effect of the second spring 633, is clamped in the side of sequence testing chip 7, sequence testing chip 7 is adjacent on retaining ring 67, deviates from even also unlikely while guaranteeing that sequence testing chip 7 is inverted.
For the ease of handling sequence testing chip 7, as preferably, as shown in figure 21, DNA sequencer in the present embodiment comprises bracing frame 84, base 8 is rotatably connected on bracing frame 84 by rotating shaft 87 as shown in figures 1 and 3, 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 21, base 8 is fixedly 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 18, for the CCD camera with the reaction warehouse for DNA sequencer of the present embodiment coupling comprise camera body 1 and be positioned at described camera body 1 for reading the fibre faceplate 4 of the optical signal that occurs in the sequencing reaction on described sequence testing chip 7, the front end that one end of fibre faceplate 4 stretches out described camera body 1 directly sees above and describes in detail with the structure of the mount pad 6 of the mount pad 6(sequence testing chip of the sequence testing chip that is positioned at DNA sequencer) 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 between 1-15 μ m.Fibre faceplate 4 directly stretches out the front end of camera body 1, does not use camera lens, fibre faceplate 4 can with sequence testing chip 7 direct-couplings, simplify light path, reduce light loss, improve the quality of order-checking signal.
As shown in Figure 18, Figure 21, in the present embodiment, as preferably, the front end of camera body 1 is connected with joint flange 2.As shown in Figure 18, Figure 19 and Figure 20, the front end of camera body 1 have for insertion be located at ring groove 66 on the mount pad 6 of sequence testing chip and with the close-fitting flange collar 21 of ring groove 66.Joint flange 2 is bolted with camera body 1, and the fibre faceplate 4 in camera body 1 is positioned at the zone that flange collar 21 surrounds, and flange collar 21 is stretched out the outside that joint flange 2 is centered around fibre faceplate 4.The end face of fibre faceplate 4 stretches out flange collar 21 certain distances, can between 1-3mm, be adjusted, in the present embodiment, the end face of fibre faceplate 4 stretches out the distance of described flange collar 21 for 2mm, so that the end face of fibre faceplate 4 can stretch in the installation cavity 69 of mount pad 6 of sequence testing chip, with sequence testing chip 7, directly contacts.The present embodiment by arrange can with the direct-coupled fibre faceplate 4 of sequence testing chip, the faint visible ray that the sequencing reaction that can make to carry out on sequence testing chip 7 produces 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 flange collar 21, makes flange collar 21 insert interior can the formation of ring groove 66 and is sealed and matched.As shown in figure 21, when the mount pad 6 the close sequence testing chip of CCD camera, be arranged on 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, ring-type flange collar 21 on joint flange 2 enters in the ring groove 66 on the mount pad 6 of sequence testing chip, flange collar 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, further guarantee that the luminous energy that sequencing reaction produces is received by fibre faceplate 4, guarantee obtaining of order-checking signal.By at camera body 1 front end, increasing the joint flange 2 with flange collar 21, and offer corresponding ring groove 66 on the mount pad 6 of sequence testing chip, thereby form the darkroom environment between the mount pad 6 of sequence testing chip and camera body 1, while guaranteeing fibre faceplate 4 coupling 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.
The structure when reaction warehouse for DNA sequencer that Figure 21 shows the present embodiment is connected with the CCD camera, structural representation when in running order for the reaction warehouse of DNA sequencer.As shown in figure 21, 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 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, between the mount pad 6 of sequence testing chip and reaction tank body 5 also for being fixedly connected with, thereby sequence testing chip 7 can drive reaction tank body 5 endwisely slipping along tail rod 9 on tail rod 9, because reaction tank body 5 is subject to the reactive force of the first spring 81 between reaction tank body 5 and base 8, between the outer face of sequence testing chip 7 and fibre faceplate 4, mutually be close to.In the outer face of sequence testing chip 7 with fibre faceplate 4 do not realize fully parallel before, due to the reactive force that is subject to the first spring 81, in the outer face of fibre faceplate 4, be close in the process of sequence testing chip 7, reaction tank body 5 is when being subject to the reactive force of the 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 sequence testing chip 7.Sequence testing chip 7 is by the adjustment on tail rod 9 is axial, 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 thering is the high parallelism precision of trying one's best between the outer face of fibre faceplate 4 and sequence testing chip 7, guarantee to greatest extent at fibre faceplate 4 when sequence testing chip 7 reads the fluorescent signal of sequencing reaction, prevented external optical pollution, thereby reduced ground unrest, realized obtaining of high s/n ratio order-checking signal.
Certainly, the above is the preferred embodiment of the present invention, should be understood that; for those skilled in the art; under the premise without departing from the principles 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 (10)
1. the reaction warehouse for DNA sequencer, is characterized in that, comprises base, carries out the reaction tank body of sequencing reaction and for the mount pad of sequence testing chip is installed;
Offer annular recesses on the surface of described reaction tank body one side;
Described mount pad is provided with the installation cavity of perforation, described sequence testing chip is arranged in an end of described installation cavity, wherein, one end of described installation cavity is provided with for locating the retaining ring of described sequence testing chip, radially be provided with the locating slot of the described inwall of a plurality of perforations on the inwall of described installation cavity along it, a plurality of described locating slots are positioned on the axial same position of described installation cavity, a plurality of described locating slots distribute along the even circumferential of described installation cavity, in each described locating slot, being equipped with slides to stretch in described installation cavity to hold described sequence testing chip along described locating slot is close on described retaining ring it and retracts in described locating slot to unclamp the slip hyoplastron of described sequence testing chip along described locating slot,
The side that described reaction tank body offers annular recesses is fixedly connected with described mount pad towards described installation cavity; Described sequence testing chip and described reaction tank body offer between the side of annular recesses and are provided with the gap of flowing through for the order-checking liquid, and described gap surrounds the sequencing reaction pond for sequencing reaction by the sealing-ring that is embedded in described groove and protrude described annular recesses;
The another side of described reaction tank body is attached on described base, and be provided with corresponding a plurality of through holes on described reaction tank body and described base, described reaction tank body is connected with described base by the tail rod through described through hole, one end of described tail rod is fixedly connected with described base, described reaction tank body by described through hole be enclosed within on described tail rod can described tail rod axially on slide and the fine motion that can make progress in the footpath of described tail rod;
Be provided with the first elastomeric element between described reaction tank body and described base.
2. the reaction warehouse for DNA sequencer as claimed in claim 1, is characterized in that, described reaction tank body comprises backboard and be located on described backboard the boss protruded to a side of described backboard, and described annular recesses is opened on the end face of described boss;
Described mount pad comprises pedestal, described installation cavity is opened on described pedestal, is fixedly connected with between described pedestal and described backboard and makes described boss stretch in described installation cavity and make between described boss and described sequence testing chip to be provided with the described gap of flowing through for the liquid that checks order;
The side that described reaction tank body is provided with backboard is attached on described base, and a plurality of described through hole of described reaction tank body setting corresponding to described base is opened on described backboard.
3. the reaction warehouse for DNA sequencer as claimed in claim 1 or 2, it is characterized in that, described the first elastomeric element is the first spring, described base is provided with cavity with the side that described reaction tank body is connected, be provided with the spring lever that one or more integral body is positioned at described cavity in described cavity, described the first spring housing is on described spring lever and stretch out described cavity;
The described through hole be located on described reaction tank body is step-like, one section of the close described base of described through hole is glissile segment, another section of described through hole is construction section, and the internal diameter of described construction section is greater than described glissile segment, and described construction section and described glissile segment junction form first step;
One end of described tail rod is the connection section be connected with described base, the other end of described tail rod is provided with for being held in the lug boss at described first step place, and the middle part of described tail rod is near the sliding part of part for sliding in described glissile segment of described lug boss;
The diameter of described glissile segment is a bit larger tham the diameter of described sliding part.
4. the reaction warehouse for DNA sequencer as claimed in claim 1 or 2, is characterized in that, described annular recesses is hexagon, and when described reaction tank body is vertically placed, described hexagonal upper end is provided with liquid outlet, and described hexagonal lower end is provided with fluid inlet.
5. the reaction warehouse for DNA sequencer as claimed in claim 1 or 2, is characterized in that, described DNA sequencer comprises the bracing frame of vertical setting, and described base is rotatably connected on support frame as described above.
6. the reaction warehouse for DNA sequencer as claimed in claim 1 or 2, is characterized in that, described locating slot is less near the size of a section of described installation cavity, is formed for the second step of spacing described slip hyoplastron; Described locating slot is provided with away from an end of described installation cavity the baffle plate be fixedly connected with described mount pad;
It is interior to hold described sequence testing chip that one end of described slip hyoplastron stretches into described installation cavity, and described slip hyoplastron is provided with and is against the protuberance on described second step when an end of described slip hyoplastron stretches out;
Be provided with between described protuberance and described baffle plate and make described slip hyoplastron stretch into the second elastomeric element in described installation cavity.
7. the reaction warehouse for DNA sequencer as claimed in claim 6, it is characterized in that, described the second elastomeric element is the second spring, described the second spring housing is on the described slip hyoplastron between described protuberance and described baffle plate, described baffle plate is provided with the through hole passed away from an end of described installation cavity for described slip hyoplastron, the inboard of described baffle plate is provided with the cover that internal diameter is greater than described through hole, the end that is formed for making described the second spring between described cover and through hole against the 3rd step.
8. the reaction warehouse for DNA sequencer as claimed in claim 1 or 2, is characterized in that, the outer side of described mount pad is provided with the ring groove that is centered around described sequence testing chip; Be connected with joint flange for the front end of CCD camera that reads the optical signal that occurs in the sequencing reaction on described sequence testing chip, the front end of described joint flange has the flange collar for inserting described ring groove and being sealed and matched with described ring groove.
9. the reaction warehouse for DNA sequencer as claimed in claim 8, it is characterized in that, described CCD camera comprise camera body and be positioned at described camera body for reading the fibre faceplate of the optical signal that occurs in the sequencing reaction on described sequence testing chip, an end of described fibre faceplate stretches out that the front end of described camera body is direct directly contacts to obtain with a side of the sequence testing chip that is positioned at described mount pad the optical signal that the sequencing reaction that carries out on the another side of sequence testing chip produces.
10. the reaction warehouse for DNA sequencer as claimed in claim 1 or 2, it is characterized in that, the zone that the annular recesses of described reaction tank body surrounds is provided with one or more temperature sensors, the side that described reaction tank body is connected with described base is provided with to the heating chamber of described reaction tank body indent, in described heating chamber, is provided with semiconductor heat booster.
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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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| CN102766574A CN102766574A (en) | 2012-11-07 |
| CN102766574B true 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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Effective date of registration: 20131223 Address after: 100101 building 7, building 1, Beichen West Road, Chaoyang District, Beijing Institute of genome, Chinese Academy of Sciences, Beijing Patentee after: Beijing Institute of Genomics, Chinese Academy of Sciences Patentee after: JILIN ZHONGKE ZIXIN TECHNOLOGY CO., LTD. Address before: 101318, building 6, B District, Shunyi District Airport Technology Park, Beijing Patentee before: Beijing Institute of Genomics, Chinese Academy of Sciences Patentee before: Semiconductor Inst., Chinese Academy of Sciences |