CN105420096B - A kind of DNA image based on reaction temperature adjustment gathers sequencing system - Google Patents
A kind of DNA image based on reaction temperature adjustment gathers sequencing system Download PDFInfo
- Publication number
- CN105420096B CN105420096B CN201510872566.8A CN201510872566A CN105420096B CN 105420096 B CN105420096 B CN 105420096B CN 201510872566 A CN201510872566 A CN 201510872566A CN 105420096 B CN105420096 B CN 105420096B
- Authority
- CN
- China
- Prior art keywords
- reaction
- base
- collection
- value
- gray value
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Sequencing system is gathered the present invention relates to a kind of DNA image based on reaction temperature adjustment, described image procossing and control system obtain profile information, m collection of illustrative plates is obtained respectively in the sampling time interval of each collection of illustrative plates Δ t, and m is even number；Collection of illustrative plates of taking pictures to same base is grouped, and is then judged by way of redundancy judges, and compares with the base profile information of standard, and tetra- kinds of base information of A, T, C, G determine the base type included in profile information.The mode of redundancy determination in the present invention, can effectively judge that collection of illustrative plates gathers accurate location of problems, conveniently take measures, and the cost of running software is relatively low, and collection of illustrative plates frequency acquisition can be arbitrarily set according to accuracy, is adapted to large-scale application；Also, the present invention to gray value when sampling, the interference of reaction warehouse temperature change is excluded, reaction warehouse temperature is maintained in default scope, it is ensured that the accuracy of sampling.
Description
Technical field
Gather and survey the present invention relates to DNA sequencing analysis field, more particularly to a kind of DNA image based on reaction temperature adjustment
Sequence system.
Background technology
In DNA sequencing technology field, integrated operation flow is described as follows：DNA sample by after broken, using building storehouse reagent
Carry out adjunction head, single-stranded capture, be bound to microballoon, microemulsion PCR amplifications, demulsification liquid, obtain the DNA texts set up on microballoon
Storehouse, the enzyme for needing library and sequencing reaction using sample-adding plate etc. are laid to the sequence testing chip with micro reaction pool, sequence testing chip
Be attached on main frame with sequencing reagent, sequencing program started according to module number and position by control computer, automate into
Row sequencing reaction, to Data Analysis Computer, computation analysis software carries out image to the data transfer of generation after completing sequencing
The work such as treatment, sequence reading, quality analysis, sequence assembly, finally give the sequence information of DNA sample.
Micro reaction pool sequence testing chip is the carrier of sequencing reaction, and DNA Beads and the various sequencings for being loaded with sequencing template are anti-
It is respectively positioned in the sequence testing chip for being carved with micro reaction pool using enzyme.
, it is necessary to process the information of single image after the IMAQ to reaction chip, especially to the letter of image
Breath accuracy is judged, it is to avoid the accuracy missing of image causes the judgement of base type to slip up；When to image measurement,
The precision of CCD camera position is the factor for influenceing image to obtain accuracy, when being obtained to image, the change of CCD camera position
Change needs adjustment.
In view of drawbacks described above, creator of the present invention passes through prolonged research and practice obtains this creation finally.
The content of the invention
It is an object of the invention to provide it is a kind of based on reaction temperature adjustment DNA image gather sequencing system, to gram
Take above-mentioned technological deficiency.
To achieve the above object, the present invention provides a kind of DNA image based on reaction temperature adjustment and gathers sequencing system, bag
Include and be provided with the reaction warehouse component of reaction chip, motion platform component, motion control unit, be arranged on motion platform component simultaneously
CCD camera, platform component and the control CCD camera controlled by motion control unit obtain image and image are carried out superfluous
The image procossing and control system of remaining treatment, wherein,
Described reaction warehouse component, in the presence of control system, reaction solution into being chemically reacted in reaction chip,
Produce visible ray；Described motion platform component, it feeds CCD camera according to the control instruction of image procossing and control system
To appropriate picture-taking position, the visible optical information to being produced in reaction chip carries out collection of taking pictures；Described platform component,
Its support described reaction warehouse component, motion platform component；
Described image procossing and control system obtain profile information, in the sampling time interval of each collection of illustrative plates Δ t
M collection of illustrative plates is obtained respectively, and m is even number；Collection of illustrative plates of taking pictures to same base is grouped, and then enters by way of redundancy judges
Row judges, and compares with the base profile information of standard, and tetra- kinds of base information of A, T, C, G determine to be included in profile information
Base type；
Described image procossing and control system include grouped element, data collection control unit, comparison module and logic
The collection of illustrative plates or image information of above-mentioned collection are carried out packet arrangement by control module, described grouped element according to two bit matrix；
The precession diagram information that m is 12 is obtained, respectively each two adjoining time, the two of gray value is organized as a two dimension
Individual dimension values, according to label 1-12, are respectively labeled as 6 groups of two dimensional gray values, and first and second gray value therein is first group, the
3rd, four gray values are second group, and the five, the six gray values are the 3rd group；Seven, the eight gray values are the 4th group, the nine, the ten gray values
Be the 5th group, the 11st, 12 gray values be the 6th group, the gray value information that the grouped element will be gathered is according to above-mentioned packet
Three groups of two dimensional gray value matrixs arrange and obtains matrix (i1,i2)、(i3,i4)、(i5,i6)、(i7,i8)、(i9,i10)、
(i11,i12)；
Include a comparator in the comparison module, its gray value collection signal to all data acquisition module memory storages
Calculation process is carried out, a registration P is calculated respectivelyn, and transmit it in the Logic control module；It is described to compare mould
Above-mentioned registration is done difference and taken absolute value by block two-by-two, after calculating result, signal registration and absolute difference is calculated
Result is transmitted to the Logic control module；The Logic control module is a MCU processors or a PLC, and it includes depositing
Storage module, is stored with the registration threshold value P of a standard base0With one group of standard value δ1And δ2；
Described data collection control unit collection registration absolute value differences are more than threshold value P0Corresponding gradation of image
Value δ, described gray value δ and standard gray angle value δ1And δ2Contrasted, if δ1＜ δ ＜ δ2, then the gray value of the image is more than
One section of preset value, need to be adjusted to the position of CCD camera；
Described Logic control module sends control instruction to motion control unit；Motion control unit controlled motor is moved
Make, motor drives leading screw to rotate, the nut and nut mounting seat on leading screw drive CCD camera movement；In the position of base sidepiece
The position of sensor real-time detection CCD camera, and transmit into motion control unit, motion control unit is in real time by positional information
Transmit into Logic control module, and compare with the default CCD camera positional information for storing in a storage module, until
CCD camera is moved to specified location；
Described Logic control module sends control instruction, electrical heating wire to the temperature control component being arranged in reaction warehouse base
Corresponding heating reduces temperature, the temperature in described temperature sensor real-time detection reaction warehouse and by real time temperature information
Transmit into Logic control module, and compare with the temperature information of memory module in it, until meeting the requirements.
Further, first group of two dimensional gray value matrix and second group of two dimensional gray value matrix, described comparing mould are obtained
(1) calculates the registration P of gray value in two groups of two dimensional gray value matrix signals to block according to the following equation21,
In formula, P21Represent the registration of every two groups of gray values, i1And i2First group of two dimensional gray value square is represented respectively
The gray value of battle array, i1Represent the sampled value of the first gray value, i2Represent the sampled value of the second gray value；i3And i4Institute is represented respectively
State second group of gray value of two dimensional gray value matrix, i3Represent the sampled value of the 3rd gray value, i4Represent the sampling of the 4th gray value
Value；T represents mean square deviation computing, and I represents integral operation.
Further, (2) are carried out according to the following equation for first group of Two-dimensional current matrix and the 3rd group of two dimensional gray value matrix
Registration calculates P31,
In formula, P31Represent the registration of every two groups of gray values, i1And i2First group of two dimensional gray value square is represented respectively
The gray value of battle array, i1Represent the sampled value of the first gray value, i2Represent the sampled value of the second gray value；i5And i6Institute is represented respectively
State the 3rd group of gray value of two dimensional gray value matrix, i5Represent the sampled value of the 5th gray value, i6Represent the sampling of the 6th gray value
Value；T represents mean square deviation computing, and I represents integral operation.
Further, (3) are entered according to the following equation for second group of two dimensional gray value matrix and the 3rd group of two dimensional gray value matrix
Row registration calculates P32,
In formula, P32Represent the registration of every two groups of gray values, i3And i4Second group of two dimensional gray value square is represented respectively
The gray value of battle array, i3Represent the sampled value of the first gray value, i4Represent the sampled value of the second gray value；i5And i6Institute is represented respectively
State the 3rd group of gray value of two dimensional gray value matrix, i5Represent the sampled value of the 5th gray value, i6Represent the sampling of the 6th gray value
Value；T represents mean square deviation computing, and I represents integral operation.
Further, the output shaft of described motor is connected with leading screw by shaft coupling, and motor is set with leading screw junction
There is support base, the end of leading screw is provided with bearing and axial positioning sleeve；
Nut is connected with leading screw, nut is arranged in nut mounting seat；The upper end of nut mounting seat connects with connecting plate
Connect, the upper end of connecting plate is connected with camera mounting seat；In bearing end, bearing friction top is set.
Further, described nut mounting seat is arranged on on the sliding block of guide rail contact, and camera is driven on sliding block
It is mobile；Position sensor mounting seat is installed in the sidepiece of base, the upper end of sensor installation seat sets contiguous block in position, even
Connect block upper end and fix position sensor.
Further, described reaction chip component includes reaction chip mounting seat, reaction warehouse base assembly, in reaction warehouse
The center section of the lateral surface of base is provided with temperature control component；
The middle part of reaction warehouse base is provided with electrical heating wire, it is arranged in the heating wire mounting groove of base；
The temperature sensor mounting groove on electrical heating wire top, is provided with temperature sensor and control assembly.
Further, reaction tank is provided with the medial surface of reaction warehouse, the surrounding of reaction tank is provided with sealing ring；
The surrounding of temperature sensor mounting groove is provided with four guide post mounting holes, upper and lower sides are provided with self adaptation bullet
Spring mounting hole；It is also equipped with being oriented to rod aperture on the cover board.
Beneficial effects of the present invention are compared with prior art：DNA image collection of the present invention based on reaction temperature adjustment
Sequencing system, described image procossing and control system obtain profile information, and the alkali that reagent place contains is included in profile information
Base type information, the information is compared with the base profile information of standard, tetra- kinds of base information of A, T, C, G, determines that collection of illustrative plates is believed
The base type included in breath.When described image procossing and control system determine information, same base is taken pictures first
Collection of illustrative plates is grouped, and is then judged by way of redundancy judges.
Above-mentioned collection data are carried out packet arrangement and obtain two dimensional gray value matrix by grouped element according to two-dimensional matrix, are easy to dress
Putting carries out software control, accuracy of judgement, quick；The each group two dimensional gray value matrix of collection is carried out into a redundancy determination, according to sentencing
Disconnected result is accordingly controlled, in the present embodiment for the redundancy of redundance judges.
The mode of redundancy determination in the present invention, can effectively judge that collection of illustrative plates gathers accurate location of problems, conveniently adopt
Measure is taken, and the cost of running software is relatively low, and collection of illustrative plates frequency acquisition can be arbitrarily set according to accuracy, is adapted to large-scale application；
Avoid erroneous judgement of the single gray value to base type again simultaneously.The present invention excludes reaction warehouse temperature when being sampled to gray value
The interference of change, reaction warehouse temperature is maintained in default scope, it is ensured that the accuracy of sampling.
Brief description of the drawings
Fig. 1 is the overall structure diagram that the DNA image based on reaction temperature adjustment of the invention gathers sequencing system；
Fig. 2 is the dimensional structure diagram of reaction warehouse component of the invention；
Fig. 3 is the structural representation of reaction chip component of the invention；
Fig. 4 is the cross section structure diagram of turnable reaction warehouse component of the invention；
Fig. 5 is the structural representation of motion platform component of the invention；
Fig. 6 is the dimensional structure diagram of the integrated tube assembly of pump valve of the invention；
Fig. 7 is the backsight structural representation of the integrated tube assembly of pump valve of the invention.
Specific embodiment
Below in conjunction with accompanying drawing, the technical characteristic above-mentioned and other to the present invention and advantage are described in more detail.
Refer to shown in Fig. 1, it is the entirety that the DNA image based on reaction temperature adjustment of the invention gathers sequencing system
Structural representation, the DNA image collection sequencing system based on reaction temperature adjustment in the embodiment of the present invention includes reaction warehouse group
Part 12, motion platform component 14, motion control unit 15, CCD camera 101, platform component 13 and image procossing and control
System processed, wherein,
Described reaction warehouse component 12, sets reaction chip component in it, in the presence of control system, reaction solution enters
Chemically reacted in reaction chip, produced visible ray；Described motion platform component 14, be fed into CCD camera suitably by it
Picture-taking position, in reaction chip produce visible optical information carry out collection of taking pictures；Described platform component 13, its branch
Support described reaction warehouse component 12, motion platform component 14；Described motion control unit, it is to CCD camera and reaction core
The operation real-time control of piece.
In embodiments of the present invention, described reaction warehouse component 12 can be installed, fixed and adjust described reaction chip,
Ensure that reaction chip is reacted in the environment of stabilization, also, reaction chip can conveniently take out and lock.
Refer to shown in Fig. 2, described reaction warehouse component 12 includes being arranged on perpendicular on described platform component 13
The straight door frame 121 for setting, is provided with turnable reaction warehouse component 124, in turnable reaction warehouse on the top in the outside of door frame 121
The lower end of component 124 is connected with turnable basement 122, and turnable basement 122 is fixed on the middle part of door frame 121, may be reversed reaction
Storehouse component 124 can rotate around described turnable basement 122, be used to take out or be put into reaction chip.
Electronics lock set 125 is additionally provided with the left of the upper end of described door frame 121, only in electronics lock set 125
During opening, turnable reaction warehouse component 124 can be rotated, reaction chip is operated.In embodiments of the present invention, if
Door frame 121 is equipped with, reaction chip and dependency structure is supported, the stability of system is substantially increased.
May be reversed reaction warehouse component 124 includes may be reversed warehouse, is provided with the center section of described turnable warehouse
Back shroud 1242, is used to encapsulate Reagent Tube；Bottom is provided with the first rotating part, it includes the first accommodating chamber 1243, is used to hold
Receive turnable basement 122, the first accommodating chamber 1243 is arranged on the centre position of bottom；It is to be symmetrical arranged in the first accommodating chamber both sides
First lug 1244, have through hole 1246 in the middle setting of first lug 1244；To connect with described turnable basement 122
Connect.Turnable basement is rotated relative to turnable warehouse in the first accommodating chamber 1243, with stability higher, while ensureing examination
Agent and the transmission of data are unimpeded.
Reagent Tube mounting hole and electric wire are provided with the upper surface of the first accommodating chamber 1243 of described turnable warehouse
Mounting hole, is provided with pipe clamp 03 in Reagent Tube mounting hole, and being passed through in pipe clamp 03 has Reagent Tube 02 and electric wire 01.Can in described
The sidepiece for overturning warehouse is additionally provided with the dead bolt mounting hole of the dead bolt 1252 in electronics lock set 125；In the presence of locking member,
Warehouse is may be reversed to be fixed together with door frame 121；Dead bolt 1252 is controlled by inductive switch 1251.In described turnable warehouse
The upper end in outside be provided with pressing piece 1241, only can make described may be reversed pressing to set by pressing piece 1241
Warehouse rotates.
Turnable basement 122 in the embodiment of the present invention has two connect bases of the second lug 1221, and second is convex
Ear 1221 is symmetricly set on a side 1223 of base, and through hole 1224 is provided with the middle of it, with described turnable storehouse
The through hole 1246 of the middle setting of the first lug 1244 on body is co-axially mounted, also, the second lug 1221 is convex installed in first
The inner side of ear 1244.
In embodiments of the present invention, door frame 121 has the structure of sidepiece baffle plate 1211 for one side, and it is integrally formed, institute
The both sides of the turnable basement 122 stated are arranged in sidepiece baffle plate 1211 and contact positioning with sidepiece baffle plate 1211；Described
One lug 1224 after being co-axially mounted with the second lug 1221, the upper-end contact of end and turnable basement 122, also, can turn over
The sidepiece of roll-over body is contacted with the medial surface of sidepiece baffle plate 1211 of door frame 121, it is ensured that may be reversed warehouse firm with turnable basement 122
It is located in sidepiece baffle plate 1211 well.
The receiving hole passed through for reaction chip component is additionally provided with door frame 121.The use of door frame, be not only able to so that
The Stability Analysis of Structures of reaction chip component, and, also cause to may be reversed warehouse and turnable basement compact conformation, it is easy to operation；CCD
Camera is more accurate with the positioning of reaction chip.The bottom of door frame 121 is provided with the first mounting hole 1210, to flat with motion
Platform component 14 is connected；The second mounting hole being connected with turnable basement sidepiece is additionally provided with the middle part of door frame.
Refer to shown in Fig. 3, reaction chip component of the invention includes reaction chip mounting seat 127, reaction warehouse base group
Part 126 and the shading positioning component 128 reaction warehouse base assembly 126 and reaction chip mounting seat 127 locked, is opened；For
Easily reaction chip can be taken out and closed, reaction chip mounting seat 127 can be relative with reaction warehouse base assembly 126
Rotation.
Refer to shown in Fig. 4, it is the structural representation of the medial surface of reaction warehouse base of the invention, in reaction warehouse
Reaction tank is provided with side, the surrounding of reaction tank is provided with sealing ring, prevent reaction reagent from leaking.In reaction warehouse base
The center section of lateral surface is provided with temperature control component, and the surrounding of temperature sensor mounting groove 1262 is provided with four guide post peaces
Dress hole 1742, self adaptation spring mounting hole 1741 is provided with upper and lower sides；It is also equipped with being oriented to rod aperture 1701 on cover plate 1263.
The middle part of reaction warehouse base is provided with electrical heating wire 1754, its heating wire mounting groove for being arranged on base
In；In the temperature sensor mounting groove on electrical heating wire top, temperature sensor and control assembly are provided with, are used to detect reaction tank
Temperature and according to setting threshold value be adjusted；In temperature sensor and control assembly 1753, it is encapsulating resin.In reaction
The avris of the interior grooves in pond is provided with sealing ring 1713.Reaction chip 04 is installed in the centre of reaction chip mounting seat, is tried
Agent is flowed through reaction chip and is reacted.
In embodiments of the present invention, Reagent Tube 02 is fixed by Reagent Tube screw 03；By the Reagent Tube that may be reversed warehouse
Mounting hole, being arranged on the middle setting of turnable warehouse has the first cavity for accommodating reagent pipeline, final coupled reaction pond body, examination
Agent pipeline by various cavitys sealing, stability is high, is hardly damaged.
Back shroud 1242 is fixed with rear side of reaction warehouse, the middle setting of back shroud 1242 has self-adaptative adjustment part
1751, in embodiments of the present invention, to may be reversed and be provided with self-adaptative adjustment part between reaction warehouse and reaction warehouse base 126
1751, the end spring of self-adaptative adjustment part 1751 and the end contact of reaction warehouse base；When being fed using CCD camera, phase
When machine is with reaction chip component touch, when camera position is adjusted, can be adjusted by the way of movable, it is now, adaptive
Answering adjustment part can change the position of reaction chip component according to the change in location of camera, it is ensured that with accuracy of detection higher,
Simultaneously, it is to avoid system is caused to damage.
In embodiments of the present invention, in order to ensure that may be reversed reaction warehouse component can coaxially move with reaction chip component,
Convenient adjustment, is additionally provided with guide post between the two, is set on reaction warehouse base and is oriented to rod aperture, and described guide post includes anti-
Storehouse locating flange is answered, to be positioned with guiding rod aperture；Positioned with the end face of guide post mounting hole by nut 1755 in the other end,
Pad is additionally provided between.Described motion platform component 14 includes that the leading screw that motor 136 is connected with motor output shaft is led
The support base and detection components, camera support component of rail assembly, support lead screw assembly.
Refer to shown in Fig. 5, the output shaft of motor 136 is connected with leading screw by shaft coupling, motor sets with leading screw junction
Support base 1361 is equipped with, the end of leading screw is provided with bearing and axial positioning sleeve.Nut is connected with leading screw, nut sets
Put in nut mounting seat；The upper end of nut mounting seat is connected with connecting plate 1012, and the upper end of connecting plate 1012 is connected with camera
Mounting seat 1011；In bearing end, bearing friction top is set.
Described nut mounting seat is arranged on the sliding block 1012 contacted with guide rail 132, and drives camera to move on sliding block
It is dynamic.Position sensor mounting seat 1351 is installed in the sidepiece of base, the upper end of sensor installation seat 1351 sets and connects in position
Block is connect, position sensor 135 is fixed in contiguous block upper end.In embodiments of the present invention, described image procossing and control system
Course of reaction in reaction warehouse is taken pictures, described CCD camera is taken pictures according to default instruction, obtains profile information, and
Profile information is transmitted and is compared and is processed into image procossing and control system.
Refer to Fig. 6-7 to show, the integrated tube assembly of pump valve of the embodiment of the present invention is by the syringe needle suction pipe 507 of downside by reagent
Sucked from the Reagent Tube on buffer solution case loading/unloading platform component, and by the selection of fluid path control unit 502, drive control valve
Component 509 and hydraulic pump 506 are selected, and reagent is entered in reagent cartridge, carry out sequencing reaction.
The integrated tube assembly of described pump valve includes pump valve integrated base plate 508, and pump valve integrated base plate 508 includes peristaltic pump
Installing plate 5011 and the fixed plate of the connection of peristaltic pump installing plate 5011, above-mentioned each plate surround a ringent body structure；At this
The centre of casing is provided with control valve group part 509.Pump valve integrated base plate 508 also include being arranged on sipper pore group on base plate,
And the control valve group part installing plate in the middle of base plate；Peristaltic pump mounting hole is provided with peristaltic pump installing plate, two are set thereon
Individual peristaltic pump 505；Interval setting multiple reagent pipe clamps and wire clamp are gone back on base plate 508.
Accordingly, control valve group part 509 is arranged on the center section of pump valve integrated base plate 508, and hydraulic pump 506 is arranged on
The sidepiece of pump valve integrated base plate 508, fluid path control unit 502 is set in the other side；Fluid path control unit 502 passes through a folding
Bent plate 503 is isolated with fluid path part.
Pump valve assemblies are arranged in the cabinet space of multiple plate compositions, are easily installed on the diverse location of tester；
In the embodiment of the present invention, the integrated tube assembly of pump valve is arranged on the upper side connecting plate of sequenator casing, and downside is installation Reagent Tube
Buffering liquid case loading/unloading platform component.
In embodiments of the present invention, in order to carry out sequencing reaction to plurality of reagents, the sipper pore group on base plate is side by side
Multiple holes are provided with, syringe needle suction pipe passes through sipper pore, the other end to connect Reagent Tube and connected with the fluid path port of corresponding control valve；
Pipe clamp is set on sipper pore, prevents syringe needle suction pipe from inclining.When in use, the examination needed for control valve control syringe needle suction pipe is chosen
Agent, is entered in reaction warehouse by the effect of syringe needle suction pipe, control valve and peristaltic pump and is reacted.Multiple pins are set on plate
Hole, selects different reagents, and selection is accurate, reliable.
The embodiment of the present invention, using the integrated tube assembly of pump valve, can simultaneously select plurality of reagents, and according to pre-
The order for first setting carries out sequencing reaction, and testing efficiency is high.
In the present invention, when the CCD camera obtains profile information, to signal waveform, within per the continuous K cycle,
Each cycle selects the n sampled point of predetermined time, at interval of time T0Sample once, continuous sampling M times；In order to ensure to sample number
According to referring to property and accuracy, within each cycle select n sampled point time interval Δ t according to the following equation (1)
Calculate,
In formula, Δ t represents the time interval of sampled point, and a is correction factor, and its size is determined by sampled point quantity, ω tables
Show the angular frequency of photographing signals, determined by the performance of CCD camera, β is initial phase angle, T represents the time of signal period, and λ is represented
The peak value of signal waveform.
Sampled through above-mentioned formula (1), when signal amplitude is bigger, sampled more intensive, the referring to property of sampled data is stronger；
Sampling is to signal data according to pre-conditioned sampling so that follow-up signal transacting data volume reduces, and mitigates the numerous of data processing
Miscellaneous computing.
Described image procossing and control system obtain profile information, and the base that reagent place contains is included in profile information
Type information, the information is compared with the base profile information of standard, tetra- kinds of base information of A, T, C, G, determines profile information
In the base type that includes.When described image procossing and control system determine information, first to the figure of taking pictures of same base
Spectrum is grouped, and is then judged by way of redundancy judges.
In embodiments of the present invention, described CCD camera is obtained respectively in the sampling time interval of each collection of illustrative plates Δ t
M (m >=6) individual collection of illustrative plates, m is even number.
In embodiments of the present invention, described image procossing and control system include grouped element, data acquisition control list
Unit, comparison module and Logic control module, described grouped element believe the collection of illustrative plates of above-mentioned collection or image according to two bit matrix
Breath carries out packet arrangement；In embodiments of the present invention, the gray value combination of two packet according to the collection of illustrative plates or image for obtaining.
In embodiments of the present invention, the precession diagram information that m is 12 is obtained, respectively each two adjoining time, as
Two dimension values of one two dimension group gray value, in the present embodiment, according to label 1-12, are respectively labeled as 6 groups of two dimensional gray values,
First and second gray value therein is first group, and third and fourth gray value is second group, and the five, the six gray values are the 3rd group；7th,
Eight gray values be the 4th group, the nine, the ten gray values be the 5th group, the 11st, 12 gray values be the 6th group, the grouped element
Three groups of two dimensional gray value matrixs arrange by the gray value information of collection according to above-mentioned packet and obtains matrix (i1,i2)、(i3,
i4)、(i5,i6)、(i7,i8)、(i9,i10)、(i11,i12)。
Above-mentioned gray value is not limited in the present invention for 12, and it can be individual for any even number m (m >=6), described point
Be grouped for two gray value informations of every adjacent domain according to above-mentioned packet mode still by group unit, is divided into m/2 two dimensional gray
Value matrix.
Above-mentioned collection data are carried out packet arrangement and obtain two dimensional gray value matrix by institute's grouped element according to two-dimensional matrix, are easy to
Device carries out software control, accuracy of judgement, quick.
The each group two dimensional gray value matrix of collection is carried out a redundancy determination by the data collection control unit, according to judgement
Result is accordingly controlled, in the present embodiment for the redundancy of redundance judges.
Include a comparator, its gray scale to all data acquisition module memory storages in the comparison module in the present invention
Value collection signal carries out calculation process, and a registration P is calculated respectivelyn, and transmit it in the Logic control module,
Now illustrated with first group of two dimensional gray value matrix and second group of two dimensional gray value matrix, the comparison module is according to following public affairs
Formula (2) calculates the registration P of gray value in two groups of two dimensional gray value matrix signals21,
In formula, P21Represent the registration of every two groups of gray values, i1And i2First group of two dimensional gray value square is represented respectively
The gray value of battle array, i1Represent the sampled value of the first gray value, i2Represent the sampled value of the second gray value；i3And i4Institute is represented respectively
State second group of gray value of two dimensional gray value matrix, i3Represent the sampled value of the 3rd gray value, i4Represent the sampling of the 4th gray value
Value；T represents mean square deviation computing, and I represents integral operation.
Similarly (3) carry out registration according to the following equation for first group of Two-dimensional current matrix and the 3rd group of two dimensional gray value matrix
Calculate P31,
In formula, P31Represent the registration of every two groups of gray values, i1And i2First group of two dimensional gray value square is represented respectively
The gray value of battle array, i1Represent the sampled value of the first gray value, i2Represent the sampled value of the second gray value；i5And i6Institute is represented respectively
State the 3rd group of gray value of two dimensional gray value matrix, i5Represent the sampled value of the 5th gray value, i6Represent the sampling of the 6th gray value
Value；T represents mean square deviation computing, and I represents integral operation.
(4) carry out registration meter according to the following equation for second group of two dimensional gray value matrix and the 3rd group of two dimensional gray value matrix
Calculate P32,
In formula, P32Represent the registration of every two groups of gray values, i3And i4Second group of two dimensional gray value square is represented respectively
The gray value of battle array, i3Represent the sampled value of the first gray value, i4Represent the sampled value of the second gray value；i5And i6Institute is represented respectively
State the 3rd group of gray value of two dimensional gray value matrix, i5Represent the sampled value of the 5th gray value, i6Represent the sampling of the 6th gray value
Value；T represents mean square deviation computing, and I represents integral operation.
Similarly, above-mentioned each group two dimensional gray value matrix is carried out into registration calculating respectively, draws each coincidence angle value.
It is 12 to be not limited to above-mentioned gray value in the present invention, and it can be individual for any even number m (m >=6), is divided into m/2
Individual two dimensional gray value matrix, carries out registration calculating between every two groups of matrixes.
Above-mentioned registration is done difference and taken absolute value by the comparison module two-by-two, after calculating result, signal is overlapped
Degree is transmitted to the Logic control module with absolute difference result of calculation.
The Logic control module is a MCU processors or a PLC, and it includes a memory module, is stored with a mark
The registration threshold value P of quasi- base0With one group of standard value δ1And δ2, the Logic control module is by the registration obtained by the calculating
Absolute value differences and registration threshold value P0Compare, if the registration absolute value differences are more than threshold value, conclude wherein two
Group gray value exceedes is expected, it is impossible to predicate same base.
By all registration absolute value differences respectively with registration threshold value P0Compare, then can conclude that a certain group of detection
Image can not meet the requirements, it is impossible to predicate same base.In the present embodiment, the registration threshold value P0Value is 0.95.
Also, when the type of base is judged, the gray value of m described image should be in default scope, described
Memory module in, four groups of gray value threshold ranges are provided with, in the case where registration requirement is met, to its affiliated base
Type decision.The mode of redundancy determination in the present invention, can effectively judge that collection of illustrative plates gathers accurate location of problems, conveniently adopt
Measure is taken, and the cost of running software is relatively low, and collection of illustrative plates frequency acquisition can be arbitrarily set according to accuracy, is adapted to large-scale application；
Avoid erroneous judgement of the single gray value to base type again simultaneously.
In the present invention, the result that DNA sequencing system can also judge according to redundancy, position and survey to CCD camera
The flow velocity of sequence reagent cartridge is changed.
Described data collection control unit collection registration absolute value differences are more than threshold value P0Corresponding gradation of image
Value δ (δ ＞ 0), described gray value δ and standard gray angle value δ1And δ2Contrasted, if δ1＜ δ ＜ δ2, then the gray value of the image
More than first paragraph preset value, at this time, it may be necessary to be adjusted to the position of CCD camera.Described Logic control module is controlled to motion
Unit processed 15 sends control instruction；The controlled motor 136 of motion control unit 15 is acted, and motor drives leading screw to rotate, on leading screw
Nut and nut mounting seat drive CCD camera movement；In the position of the position sensor real-time detection CCD camera of base sidepiece,
And transmit into motion control unit 15, motion control unit 15 is in real time transmitted into Logic control module positional information, and
Compare with storage default CCD camera positional information in a storage module, until CCD camera is moved to specified location.
After the completion of position adjustment, described CCD camera proceeds to take pictures to reaction chip, and reacquires gray value and registration
Value.
The end spring of self-adaptative adjustment part 1751 and the end contact of reaction warehouse base；When being fed using CCD camera,
When camera is with reaction chip component touch, when camera position is adjusted, can be adjusted by the way of movable, it is ensured that reaction
Chip is in position.Meanwhile, described Logic control module sends control to the temperature control component being arranged in reaction warehouse base
System instruction, the heating accordingly of electrical heating wire 1754 or reduction temperature, in described temperature sensor real-time detection reaction warehouse
Temperature and by real time temperature information transfer to Logic control module, and compared with the temperature information of memory module in it
It is right, until meeting the requirements.
The embodiment of the present invention excludes the interference of reaction warehouse temperature change when being sampled to gray value, by reaction warehouse temperature dimension
Hold in default scope, it is ensured that the accuracy of sampling.
The center section of the lateral surface of reaction warehouse base is provided with temperature control component, in temperature sensor mounting groove 1262
Surrounding is provided with four guide post mounting holes 1742, and upper and lower sides are provided with self adaptation spring mounting hole 1741；In cover plate 1263
On be also equipped be oriented to rod aperture 1701.
The middle part of reaction warehouse base is provided with electrical heating wire 1754, its heating wire mounting groove for being arranged on base
In；In the temperature sensor mounting groove on electrical heating wire top, temperature sensor and control assembly are provided with, are used to detect reaction tank
Temperature and according to setting threshold value be adjusted；In temperature sensor and control assembly 1753, it is encapsulating resin.
Described gray value δ and standard gray angle value δ1And δ2Contrasted, if gray value δ ＞ δ2, then gray value is more than second
Section preset value is, it is necessary to adjust the speed of agent delivery.Described Logic control module sends control and refers to fluid path control unit 502
Order, described fluid path control unit 502 sends control instruction to drive control valve module 509, and the rotating speed of hydraulic pump 506 accordingly changes
Become, the rotating speed of peristaltic pump 505 accordingly changes, Reagent Tube 02 by may be reversed warehouse Reagent Tube mounting hole, be arranged on it is turnable
The middle setting of warehouse has the first cavity for accommodating reagent pipeline, by reagent delivery to reaction chip.Examination after being adjusted
During tachometer value of the agent flow velocity in default memory module.Now, CCD camera proceeds to take pictures to reaction chip, and
Reacquire gray value and coincidence angle value.
Above-mentioned detailed description is directed to illustrating for one of them possible embodiments of the present invention, and the embodiment is simultaneously not used to
The scope of the claims of the invention is limited, all equivalence enforcements or change without departing from carried out by the present invention are intended to be limited solely by the technology of the present invention
In the range of scheme.
Claims (2)
1. a kind of DNA image based on reaction temperature adjustment gathers sequencing system, it is characterised in that including being provided with reaction chip
Reaction warehouse component, motion platform component, motion control unit, be arranged on motion platform component and by motion control unit control
The CCD camera of system, platform component and control CCD camera obtain image and the image procossing of redundancy treatment are carried out to image
And control system, wherein,
Described reaction warehouse component, in the presence of control system, reaction solution is produced into being chemically reacted in reaction chip
Visible ray；Described motion platform component, be fed into for CCD camera according to the control instruction of image procossing and control system suitable by it
When picture-taking position, in reaction chip produce visible optical information carry out collection of taking pictures；Described platform component, its branch
Support described reaction warehouse component, motion platform component；
The described reaction warehouse component for being provided with reaction chip includes reaction chip mounting seat, reaction warehouse base assembly, in reaction
The center section of the lateral surface of orlop seat is provided with temperature control component；
The middle part of reaction warehouse base is provided with electrical heating wire, it is arranged in the heating wire mounting groove of base；Add in electricity
In the temperature sensor mounting groove on heated filament top, temperature sensor and control assembly are provided with；
Described image procossing and control system obtain profile information, in the sampling time interval of each collection of illustrative plates Δ t respectively
M collection of illustrative plates is obtained, m is even number；Collection of illustrative plates of taking pictures to same base is grouped, and is then sentenced by way of redundancy judges
It is fixed, and compare tetra- kinds of base information of A, T, C, G with the base profile information of standard, determine the base included in profile information
Type；
Described image procossing and control system include grouped element, data collection control unit, comparison module and logic control
The collection of illustrative plates or image information of above-mentioned collection are carried out packet arrangement by module, described grouped element according to two-dimensional matrix；
The precession diagram information that m is 12 is obtained, respectively each two adjoining time, as two dimensions of two dimension group gray value
Value, according to label 1-12, is respectively labeled as 6 groups of two dimensional gray values, and first and second gray value therein is first group, third and fourth ash
Angle value is second group, and the five, the six gray values are the 3rd group；Seven, the eight gray values are the 4th group, and the nine, the ten gray values are the 5th
Group, the 11st, 12 gray values be the 6th group, the grouped element will gather gray value information according to above-mentioned packet by 6 groups
Two dimensional gray value matrix arrange and obtains matrix (i1,i2)、(i3,i4)、(i5,i6)、(i7,i8)、(i9,i10)、(i11,i12)；
Include comparator in the comparison module, its gray value collection signal to all data acquisition module memory storages is transported
Calculation is processed, and registration P is calculated respectivelyn, and transmit it in the Logic control module；The comparison module will be above-mentioned
Registration does difference and takes absolute value two-by-two, after calculating result, signal registration and absolute difference result of calculation is transmitted
In to the Logic control module；The Logic control module be microcontroller (MCU) or programmable logic controller (PLC) (PLC),
It includes memory module, is stored with the registration threshold value P of standard base0With one group of standard value δ1And δ2；
Described data collection control unit collection registration absolute value differences are more than threshold value P0Corresponding image intensity value δ, institute
The gray value δ for stating and standard gray angle value δ1And δ2Contrasted, if δ1＜ δ ＜ δ2, then the gray value of the image is pre- more than first paragraph
If value, need to be adjusted to the position of CCD camera；
Described Logic control module sends control instruction to motion control unit；Motion control unit controlled motor is acted, electricity
Machine drives leading screw to rotate, and the nut and nut mounting seat on leading screw drive CCD camera movement；In the position sensor of base sidepiece
The position of real-time detection CCD camera, and transmitting into motion control unit, motion control unit in real time by positional information transmit to
In Logic control module, and compare with the default CCD camera positional information for storing in a storage module, until CCD phases
Machine is moved to specified location；
Described Logic control module sends control instruction to the temperature control component being arranged in reaction warehouse base, and electrical heating wire is corresponding
Heating or reduce temperature, temperature in described temperature sensor real-time detection reaction warehouse and by real time temperature information transfer
Into Logic control module, and compare with the temperature information of memory module in it, until meeting the requirements；
The output shaft of described motor is connected with leading screw by shaft coupling, and motor is provided with support base with leading screw junction, in silk
The end of thick stick is provided with bearing and axial positioning sleeve；
Nut is connected with leading screw, nut is arranged in nut mounting seat；The upper end of nut mounting seat is connected with connecting plate, even
The upper end of fishplate bar is connected with camera mounting seat；In bearing end, bearing friction top is set.
2. the DNA image based on reaction temperature adjustment according to claim 1 gathers sequencing system, it is characterised in that institute
The nut mounting seat stated be arranged on the sliding block of guide rail contact on, and on sliding block drive camera movement；Pacify in the sidepiece of base
Equipped with position sensor mounting seat, in position, the upper end of sensor installation seat sets contiguous block, and contiguous block upper end is fixed with position
Sensor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510872566.8A CN105420096B (en) | 2015-12-02 | 2015-12-02 | A kind of DNA image based on reaction temperature adjustment gathers sequencing system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510872566.8A CN105420096B (en) | 2015-12-02 | 2015-12-02 | A kind of DNA image based on reaction temperature adjustment gathers sequencing system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105420096A CN105420096A (en) | 2016-03-23 |
| CN105420096B true CN105420096B (en) | 2017-06-23 |
Family
ID=55498640
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510872566.8A Active CN105420096B (en) | 2015-12-02 | 2015-12-02 | A kind of DNA image based on reaction temperature adjustment gathers sequencing system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105420096B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105400689B (en) * | 2015-12-02 | 2017-06-23 | 北京中科紫鑫科技有限责任公司 | A kind of image processing system for being applied to DNA sequencing |
| US11288576B2 (en) | 2018-01-05 | 2022-03-29 | Illumina, Inc. | Predicting quality of sequencing results using deep neural networks |
| AU2019205267B2 (en) * | 2018-01-05 | 2021-05-27 | Illumina, Inc. | Predicting reagent chiller instability and flow cell heater failure in sequencing systems |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102703302B (en) * | 2012-05-24 | 2013-11-27 | 中国科学院北京基因组研究所 | Installing seat for automatically clamping sequencing chip |
| CN102703311B (en) * | 2012-05-24 | 2013-08-21 | 中国科学院北京基因组研究所 | Reaction bin for adaptive regulation of DNA (Deoxyribose Nucleic Acid) sequencer |
| CN102766574B (en) * | 2012-05-24 | 2013-12-25 | 中国科学院北京基因组研究所 | Reaction chamber for DNA sequenator |
| CN102703312B (en) * | 2012-05-24 | 2014-03-19 | 中国科学院北京基因组研究所 | DNA (Deoxyribose Nucleic Acid) sequencer |
| CN105400689B (en) * | 2015-12-02 | 2017-06-23 | 北京中科紫鑫科技有限责任公司 | A kind of image processing system for being applied to DNA sequencing |
-
2015
- 2015-12-02 CN CN201510872566.8A patent/CN105420096B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105420096A (en) | 2016-03-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105524827B (en) | A kind of DNA image with linkage adjustment gathers sequencing system | |
| CN105319156B (en) | It is a kind of that sequencing system is gathered based on the DNA image that CCD camera is adjusted | |
| CN105420096B (en) | A kind of DNA image based on reaction temperature adjustment gathers sequencing system | |
| CN105296662B (en) | A kind of DNA image collection sequencing system based on reagent self-adaptative adjustment | |
| CN110238633B (en) | Novel auto-screwdriving machine towards flexible manufacturing | |
| CN105400689B (en) | A kind of image processing system for being applied to DNA sequencing | |
| CN103868470A (en) | Curvature detection device and method for drill rod | |
| CN205301888U (en) | Confirm to be used for equipment and device and operating parameter generater of operating parameter of pump sending unit of well | |
| CN105733939B (en) | Constant temperature rotary device | |
| CN110001840A (en) | A kind of double-wheel self-balancing vehicle motion control method under various road conditions of view-based access control model sensor | |
| CN111721823A (en) | Plant nutrient real-time detection system based on electrophoresis microfluidic technology | |
| CN108332655A (en) | A kind of laser measuring device for measuring and measurement method | |
| CN105316228B (en) | A kind of box DNA sequencer with straight-line feed CCD camera | |
| CN1837798A (en) | Measurer for oceanic sediment thermal conductivity in-situ measuring system | |
| CN104280033B (en) | Indexing mechanism based on strapdown inertial navigation system and electromagnetic lock fault detection method | |
| CN105806217B (en) | Single stay formula measuring system and method for the measurement of object space position | |
| CN208443608U (en) | A kind of sewage sampling device for waste water control | |
| CN210465280U (en) | Battery appearance detection device | |
| CN104062692B (en) | High-precision seabed terrestrial heat flow detection device | |
| CN208109136U (en) | A kind of infrared sensor detection device | |
| CN205786275U (en) | A kind of device for detecting safety of foods | |
| CN203685161U (en) | Panoramic drilling image obtaining system for power survey | |
| CN105627988B (en) | The detection plate inclination checking method and system of a kind of rotor | |
| CN105368707B (en) | DNA sequencer with CCD camera adjusted in single-dimensional mode | |
| JP5663328B2 (en) | Nucleic acid analyzer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |