CN1858201A - Controling device and method for polymerase chain reaction chip array - Google Patents
Controling device and method for polymerase chain reaction chip array Download PDFInfo
- Publication number
- CN1858201A CN1858201A CN 200610043243 CN200610043243A CN1858201A CN 1858201 A CN1858201 A CN 1858201A CN 200610043243 CN200610043243 CN 200610043243 CN 200610043243 A CN200610043243 A CN 200610043243A CN 1858201 A CN1858201 A CN 1858201A
- Authority
- CN
- China
- Prior art keywords
- chip
- temperature
- pcr
- interchanger
- pcr chip
- 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.)
- Granted
Links
Images
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Control Of Temperature (AREA)
Abstract
The present invention discloses a kind of control device and method for PCR chip array. The PCR chip array without built-in heater and sensor is set on a heat exchanger; four liquid storages store liquid of different temperature; and circulating pumps and eight solenoid valves make liquid of proper temperature to circulate inside the heat exchanger for the chips get required reaction temperature. Over the chip array, is the flexible printed circuit board with heaters and sensors corresponding to the chips for the dynamic trimming of the temperature of the chips. The upper surface of the heat exchanger is divided into several temperature zones, and each of the temperature zones has one chip selected as the template chip with built-in sensor and filled liquid and with the measured temperature value used as the feedback signal for controlling the temperature of the chips in the said zone. One computer with interface circuit is used for the feedforward-cascade control and statistics process.
Description
Technical field
The present invention relates to a kind of control device and control method of biochip, relate in particular to the control device and the control method of a kind of PCR chip (polymerase chain reaction chips, pcr chip) array.
Background technology
The polymerase chain reaction belongs to the pulsating Enzymology method of external rapid amplifying specific DNA, is widely used in molecular biological every field.Its ultimate principle is to utilize archaeal dna polymerase to depend on the feature of dna profiling, the dna segment both sides sequence complementary primer a pair of and to be amplified at external use brings out polyreaction, this chain DNA is earlier through high-temperature denatured (representative temperature is 95 ℃), at low temperature (55 ℃ of representative temperatures) and primer annealing, extend in middle temperature (72 ℃ of representative temperatures) more then.This process circulates repeatedly and carries out, and generally needs 20-40 cycle, and the multiplication of specific DNA quantity index access rule is convenient to further dna molecular be carried out analyzing and testing.General equipment is desk-top pcr amplification instrument at present, can increase simultaneously to the dozens of sample, and the test of once increasing needs a few hours.
Along with the rise of micro electro mechanical system (MEMS) technology, the pcr chip that is based upon on semi-conductor and the Micrometer-Nanometer Processing Technology basis has become the research focus.Because the pcr chip volume is small, quality is minimum, specific surface area is big, can be from remedying the deficiency of desk-top pcr amplification instrument in essence, reach temperature rate height (can reach per second tens degree), loop cycle is short, accuracy of temperature control is high, Dynamic Repeatability is good purpose.The general pcr chip that uses can be finished once amplification test in 30 minutes.The outstanding advantage of another of pcr chip is to save precious sample, and required sample is only for counting microlitre to tens of microlitres." micro-system design " ((U.S.) Sen Tuliya (Senturia, S.D.) work; Liu Zewen etc. translate, the Electronic Industry Press, 2004) two big classes of pcr chip have been set forth, one class is continuous flowing type pcr chip (or being called spatial domain formula pcr chip), another kind of is little reactive tank pcr chip (or being called time domain formula pcr chip, batch type pcr chip), two class pcr chips respectively have the advantage of oneself, and are on the knees of the gods now as for which kind of dominate in the market in future.The present invention is directed to little reactive tank pcr chip.
" micro-system design " also set forth a kind of temperature-controlled process of little reactive tank pcr chip, and it is integrated at chip internal with well heater, transmitter, from big inertia, the large time delay problem that has solved macroscopical temperature parameter in essence.But the chip structure complexity, production process is many, production cost is high, yield rate is low, causes the chip price height, and general each one or several sample that can only increase, and present stage is difficult to substitute conventional pcr amplification instrument on a large scale; " Sensors and Actuators B " the 9th phase in 2000 " A rapidmicro-polymerase chain reaction system for hepatitis C virus amplification " literary composition discloses the temperature-controlled process and the control device of another kind of little reactive tank pcr chip, itself does not contain well heater, transmitter its chip, but is finished the PCR temperature cycle of a chip by the temperature control unit of outside.Used heating, refrigeration device are the Peltier device, and the temperature control strategy is the control of ratio---integration---differential, i.e. PID control.This pcr chip is simple in structure, has simplified production technique, has reduced production cost, has also improved the yield rate and the reliability of chip simultaneously.But owing to the temperature control unit that only relies on the outside macroeconomic significance, though the thermal capacity of chip self is very low, specific surface area is very high, but because big inertia, the large time delay of outside temperature control unit still exist, temperature controlled rapidity, accuracy all are restricted, heating-cooling speed is respectively 4 ℃/s, 2.2 ℃/s, and also only can carry out the temperature cycle of a chip at every turn, compare with conventional pcr amplification instrument and still to lack clear superiority, the existence of the problems referred to above is that little reactive tank pcr chip does not obtain one of major reason of large-scale promotion as yet.
Summary of the invention
Purpose of the present invention is exactly in order to solve existing little reactive tank pcr chip in the problem that exists aspect the temperature control.The invention provides a kind of new control method and form control device, make many low-cost little reactive tank pcr chips that do not contain well heater, transmitter itself be subjected to temperature control fast, accurately simultaneously according to this method.
For achieving the above object, the present invention has adopted following technical scheme: a kind of control device of PCR chip array, it comprises interchanger, place the PCR chip that has at least one microcavity at the interchanger upper surface, it is pcr chip, it is characterized in that: described pcr chip has a plurality of, and they constitute the pcr chip array at the interchanger upper surface; Above the pcr chip array, flexible printed circuit board is set, on flexible printed circuit board, is manufactured with and corresponding well heater of chip array and sensor array; Simultaneously, the interchanger upper surface is divided into some warm areas, one group of pcr chip in the corresponding pcr chip array of each warm area, in each warm area, select a pcr chip as observing and controlling model chip, in model chip microcavity, place micro temperature sensor and be full of the liquid that has approximate thermal characteristic with the PCR reaction solution; Electric components such as temperature sensor, the well heater on the flexible printed circuit board and sensor array in the various kinds board chip are connected with computer control system by interface circuit respectively.
Be shaped on a plurality of unit that constitute by well heater and transmitter on the described flexible printed circuit board, the corresponding pcr chip in each unit, these unit constitute one and corresponding well heater of pcr chip array and sensor array; Flexible printed circuit board is placed in the top of pcr chip array and contacts with each pcr chip.
Is mobile heat exchange medium in the described interchanger, interchanger is the molectron of thermally conductive material and lagging material, and bottom and heat exchange medium gangway are lagging material, and all the other are thermally conductive material, and thermally conductive material forms some grooves and contacts with the heat exchange medium; The interchanger upper surface is divided into several warm areas, with the homogeneity of guaranteeing each warm area upper surface temperature in certain allowed band; Fluid intake place at each warm area is provided with at least one temperature sensor; Corresponding some the pcr chips of the interchanger upper surface of each warm area, wherein one for observing and controlling model chip all the other are the actual chip that carries out the PCR reaction; Interchanger is connected with four liquid-storing boxs, recycle pumps by pipeline, pipeline is provided with eight magnetic valves, be respectively equipped with temperature sensor, heating unit or cooling module in four liquid-storing boxs, these electrical elements all are connected with computer control system by interface circuit, each magnetic valve carries out break-make and switches under computer control, heat exchange medium in the corresponding liquid-storing box is circulated by interchanger, and the heat exchanger surface temperature reaches the numerical value that needs rapidly.
A kind of control method of PCR chip array, this method is:
Place at the interchanger upper surface and somely to have microcavity but do not contain the pcr chip of well heater, transmitter, form array; The flexible printed circuit board that will have the array of being made up of well heater and sensor unit is placed on pcr chip array top and contact with it, and each well heater and sensor unit are corresponding one by one with each pcr chip;
Under the effect of computer control system, each pcr chip is by the temperature of interchanger upper surface acquisition near polymerase chain reaction ideal temperature value; Computer control system is gathered each warm area heat exchanger entrance place's heat exchange medium temperature, model chip upper surface temperature, model chip internal fluid temperature, implement the compound control strategy of feedforward-tandem, corresponding well heater on the control flexible printed circuit board, to corresponding pcr chip heating, make it obtain temperature value accurately;
Gather the poor of each chip upper surface temperature and its model chip upper surface temperature simultaneously, implement statistical Process Control, whether be in standard state to monitor each chip thermal contact resistance.
Described feedforward-tandem control mode is: each warm area inlet heat exchange medium temperature detection signal inputs to computer control system in the interchanger, this signal after the given signal generator conversion that inside computer system forms via software on the one hand as the setting signal that is the principal controller that forms by software equally; Mutually comprehensive via the dynamic Feedforward link of software formation on the other hand with the output signal of this principal controller; Detect various kinds board chip upper surface temperature signal simultaneously and form interior ring as the feedback signal of corresponding warm area submaster controller, submaster controller is also formed by software; The temperature of various kinds board chip internal liquid is formed outer shroud as the feedback signal of corresponding warm area principal controller; The submaster controller output signal is through putting on each well heater on the pairing flexible printed circuit board of this group pcr chip after the power magnification simultaneously, to finish each pcr chip accuracy control over temperature.
Described statistical Process Control is, with the difference of chip upper surface temperature and this chip warm area model of living in chip upper surface temperature as monitoring parameter, carry out monodrome-moving range control, promptly the dispersiveness of thermal contact resistance between chip upper surface and the flexible printed circuit board has been carried out statistical Process Control, indirectly the dispersiveness of the upper and lower surperficial contact pressure of chip is carried out statistical Process Control, be equivalent to the dispersiveness of thermal contact resistance between chip lower surface and the interchanger has also been carried out statistical Process Control; When system works in statistics slave mode following time, can think the dispersiveness of each chip thermal contact resistance within allowed band, the model chip has sufficient representativeness; When the influence owing to abnormal factors makes certain chip monitoring parameter break away from the statistics slave mode, think that then the thermal contact resistance of this chip has obviously departed from normal value, the model chip no longer possesses the representativeness to this chip, and computer system provides corresponding caution.
Useful result of the present invention is: some pcr chips are placed on the interchanger upper surface of a particular design, form array; With certain suitable liquid (as pure water) is the heat exchange medium, and by recycle pump, solenoid control, the heat exchange medium that makes relevant temperature as required is at the interchanger internal recycle; System contains four liquid storage containers, wherein the fluid temperature in three containers be controlled in respectively with the corresponding temperature of three characteristic temperatures in polymerase chain reaction (55 ℃, 72 ℃, 95 ℃) on; Store cooling fluid in the 4th container, be used for accelerating temperature-fall period from high-temperature denatured to stress relief annealed.For each little reactive tank pcr chip a heating and sensing unit are set, so that provide the control of temperature accurately and fast to each pcr chip, each heating and sensing unit are produced on same the flexible printed circuit; Homogeneity and accuracy requirement according to heat exchanging device surface temperature are carried out rationalization partition with the interchanger upper surface, think that the temperature in each zonule is identical, are called an isothermal region.In each isothermal region, selection is positioned at the intermediary pcr chip as observing and controlling model chip, in its microflute, place a micro temperature sensor (as micro thermocouple) and be full of certain and the PCR reaction solution has the liquid (as distilled water) of close thermal characteristic, think that under the identical situation of well heater and transmitter heating unit input electric power fluid temperature is identical in each chip of isothermal region.Do not carry out the polymerase chain reaction in the observing and controlling model chip, finish in remaining chip the polymerase chain reaction, but do not place micro temperature sensor in these chips.When carrying out the polymerase chain reaction of dissimilar or different batches, only need to change remaining chip, need not change observing and controlling model chip, observing and controlling model chip can be considered the intrinsic configuration of TT﹠C system.The configuration of chip temperature probe and some operation have correspondingly not only been simplified in the proposition of observing and controlling model chip notion, thereby and because it is the fixed configurations of TT﹠C system people are had ready conditions to its accurate demarcation; The temperature of each observing and controlling model chip of computer-automatic collection forms each control output by the expectant control strategy, and this output not only puts on corresponding observing and controlling model chip, and puts on other chip that is in same isothermal region with this model chip simultaneously.System's principal feature is " macroscopical temperature adjusting, microcosmic temperature correction, utilize the model chip, form compound control ".
Description of drawings
Fig. 1 is a structural representation of the present invention;
Fig. 2 is the inner little reactive tank pcr chip synoptic diagram that does not contain well heater, transmitter;
Fig. 3 is based on the well heater of flexible PCB plate and sensor unit structure synoptic diagram;
Fig. 4 is a kind of pcr chip array temperature hardware control system configuration based on industrial computer;
Fig. 5 is the structural representation of a warm area on the special-purpose interchanger;
Fig. 6 is the diagrammatic cross-section of Fig. 5;
Fig. 7 is the compound control block diagram of the feedforward-tandem of chip.
Wherein, 1, interchanger, 2, flexible printed circuit board, 3, pcr chip, 4, recycle pump, 5, liquid-storing box, 6, magnetic valve, 7, well heater, 8, transmitter.
Embodiment
Below in conjunction with drawings and Examples to further detailed explanation of the present invention.
Among Fig. 1,3 one-tenth arrays of pcr chip be placed on interchanger 1 upper surface, the chip array top covers flexible printed circuit board 2, interchanger 1 is connected by pipeline and recycle pump 4, four liquid-storing boxs 5, also is provided with eight magnetic valves 6 on pipeline.
A unit of well heater on the flexible printed circuit board 2 and sensor array as shown in Figure 3.Flexible printed circuit board 2 adopts flexible substrate to make, and its principal feature is flexible folding, and the distribution high conformity can easily be installed at three-dimensional space.Adopt and the corresponding single-sided flexible printed circuit board (PCB) of interchanger 1 upper surface geometrical dimension, utilizing it on Copper Foil to have certain resistance and this resistance and temperature has the principle of definite corresponding relation to heat and sensing unit at this plate and the pairing position making of each chip.Because well heater 7 need be drawn corresponding electric power from power supply according to temperature controlled requirement, inevitably there is self-heating effect, and the size of this self-heating effect fluctuates, if well heater 7 is used as temperature sensor simultaneously, its temperature measurement accuracy certainly will will be subjected to the influence of self-heating effect.Well heater 7 and transmitter 8 are designed respectively, 8 of transmitters need the excitation of 5mA constant current source for this reason, and well heater 7 working current maximums can reach 300mA.The cross-sectional dimension of flexible printed circuit board tinsel cord is all wide 55um, thick 35um.Each well heater 7 resistance is about 3.3 ohm in the time of 20 ℃, and the resistance of each transmitter 8 is about 6.5 ohm.Flexible printed circuit board 2 is placed in the top of pcr chip 3 arrays and contacts, so that temperature correction at each chip is provided with it.Use elastic element and thermal conductive insulation glue in order to good thermo-contact between each well heater 7 and transmitter 8 and each pcr chip 3 on the assurance flexible printed circuit board, the dispersiveness of thermal contact resistance is limited within the acceptable range.Because sophisticated flexible PCB plate production technology is arranged at present, the consistent performance of each heating and sensing unit accesses assurance.Flexible printed circuit board base material thermal conductivity is low, between each well heater that is brought by it, intercoupling between each chip can ignore.
Based on the computer control system Hardware configuration of industrial computer as shown in Figure 4.Macroscopic concentrates temperature-control device to be made up of a PC bus industrial control computer, some corresponding interface integrated circuit boards, 8 direct-acting electromagnetic valves 6,4,4 2L liquid-storing box of recycle pump 5, a high-performance heat exchanger 1.Heat exchange medium in the liquid-storing box is a pure water.Wherein 3 each built-in 500W electrical heating wires of liquid-storing box and K type thermocouple temperature sensor, by the PC bus industrial control computer with its temperature be controlled at, low-temperature annealing high-temperature denatured respectively with the PCR reaction process, middle temperature is extended on three corresponding temperature of characteristic temperature.The 4th liquid-storing box 5 stores water coolant, can lower the temperature rapidly to guarantee interchanger 1.Can use artificial method on the rocks to form mixture of ice and water, also can utilize a portable compressor to realize.Industrial control computer by the interface integrated circuit board, is constantly controlled 8 magnetic valves 6 according to the needs of pcr chip 3 reaction process, and the water of corresponding water tank is circulated by interchanger 1, and interchanger 1 surface temperature reaches corresponding numerical value rapidly.By to liquid-storing box 5 temperature controlling, make interchanger 1 upper surface steady temperature hang down 2 ℃-3 ℃ than chip is temperature required, difference is replenished by the well heater 7 of each chip top.Because 7 need of each well heater provide 2 ℃-3 ℃ temperature rise, so accuracy, rapidity that chip temperature is controlled just realize than being easier to.Each well heater 7 also is computer-controlled by same.PC bus industrial computer system by main frame, CRT, two 2 groups 8 tunnel intellectual thermopair input template IPC5455, four photoelectricity isolate 32 tunnel 8 A/D plate IPC5482,32 road photoelectric switch amount output template IPC5373 and homemade resistance temperature detector signal regulating panel, the solid state relay plate is formed.
Among Fig. 5, Fig. 6, interchanger 1 heat exchange rate should adapt with the heat conduction time constant of pcr chip 3, and temperature homogeneity preferably will be arranged.Interchanger 1 upper surface adopts the high red copper material of thermal conductivity for placing effective heat-transfer surface of pcr chip 3, opens several through slots so that expand heat interchanging area, reduce the red copper quality, improve its heat exchange rate on the red copper material.For reducing calorific loss, guarantee heat exchange rate, the low plastic material of the rest part of interchanger (bottom, gangway etc.) employing thermal conductivity.It is 2 meters that the active path of fluid in interchanger is about, wide about 60 millimeters.Detect through infra red thermograph, the heat exchanger surface temperature rise rate can reach 12 ℃/s, and rate of temperature fall can reach 10 ℃/s, and heat exchanger surface temperature extreme difference is in 2.0 ℃ during same temperature fluid stable circulation.But in two kinds of temperature fluid alternative dynamic processes, because fluidic transmission lag, heat exchanger surface temperature extreme difference can be up to 9.3 ℃, heat exchanger surface is distributed according to its dynamic temperature for this reason and be divided into 12 warm areas, Fig. 6 is the structural representation of a warm area, can guarantee less than 0.7 ℃ through repetition test temperature extreme difference in each warm area in dynamic process.Establish one of K type thermocouple temperature sensor at the fluid intake place of each warm area, it has also measured the fluid outlet temperature of a warm area when measuring this warm area fluid intake temperature.Each warm area interchanger upper surface can be installed 10 pcr chips, and 120 pcr chips then can be set altogether, and wherein 12 are observing and controlling model chip, are arranged on position shown in the dash area among Fig. 5, and 108 are the actual chip that carries out the PCR reaction.
Fig. 7 is the compound control strategy synoptic diagram of the feedforward-tandem of chip.For the reaction solution in each pcr chip, there are two control inputs, first is that water arrives reaction solution again through interchanger, silicon substrate, it is formed according to PCR process control pump, valve automatically by computer; Second is that well heater, cover glass on the flexible PCB plate arrives reaction solution again.The overwhelming majority of pcr chip institute energy requirement is from interchanger in the temperature-rise period, and 7 of well heaters provide few part, plays " correction ", " picking up the neglected and to supply the deficient "; The pcr chip energy stored is distributed through water from interchanger substantially again in the temperature-fall period.To each pcr chip itself, first input is actually can survey and uncontrollable, has only second input controlled, but second input must adapt with first input again, coordinate mutually.For this reason in the processing of control strategy, artificial regards the dual input system as single input system, regard first input as " interference ", but the given signal of the temperature of chip is produced with the PCR process jointly by first control input, tandem-compound control strategy feedovers in application, chip is carried out the control of PCR temperature, formation temperature stochastic system.For example, when COMPUTER DETECTION when certain isothermal region interchanger water temperature begins to heat up by 72 ℃, illustrate that the middle temperature extension stage of chip finishes, should enter the high-temperature denatured stage as early as possible, so at once the set-point of this isothermal region chip temperature is made as 95 ℃.Why not the desired temperature of all chips in the change system simultaneously is because water mobile in interchanger exists transmission lag.Given signal generator, master loop controller, subloop controller, dynamic Feedforward link, PWM generator etc. are all realized by software by computer among Fig. 7.The controlled target of system be each isothermal region chip temperature want " fast, accurate and stable " tracking its give fixed temperature.Owing to for fixed temperature at varying cyclically, so this is a stochastic system.The interior ring of cascade control system is the chip upper surface temperature control loop, and outer shroud is a chip internal fluid temperature gate ring.The transport function structure that outputs to chip 3 upper surface temperature from the subloop controller is taken as:
T wherein
1For well heater and and chip between the concentrated expression of interface resistance, thermal capacitance, τ
1Lag behind K by well heater and the heat transfer of chip interface
1Be the static scale-up factor between well heater electric power and the interface temperature.Transport function structure from chip 3 upper surface temperature to the chip sample temperature is taken as:
T wherein
2Concentrated expression for chip self thermal inertia and sample thermal inertia; The transmission lag of chip silicon structure part is a Millisecond, can ignore, so τ
2Comprise that the heat transfer hysteresis of chip glass cover plate, the heat transfer hysteresis of sample and the detection of sample temperature lag behind K
2Be static scale-up factor.The transport function structure of fluid temperature is taken as in from each isothermal region inlet heat exchange medium temperature of interchanger to chip:
T wherein
3Be the interchanger time constant.The transmission lag of chip silicon structure part is a Millisecond, can ignore; The detection hysteresis of interchanger water temperature and the detection of sample temperature lag behind and cancel each other, so can think τ
3Only comprise that heat transfer hysteresis, interchanger and the transmission lag of chip interface of the transmission lag of water in interchanger, interchanger itself, the heat transfer of chip sample lag behind.Through step response method test, T
1=0.5s, T
2=3.1s, T
3=1.1s, τ
1=0.1s, τ
2=1.1s, τ
3=1.36s.Can determine that in view of the above the transport function of dynamic Feedforward link and the structure of major and minor controller reach roughly parameter.The concrete parameter of major and minor controller is finally adjusted by the scene and is finished.In ring time constant, time lag all very little, working speed is fast, formation is to the quick stochastic system of principal controller output signal, make the chip upper surface temperature reach needed value rapidly, and can eliminate the various disturbances that enter inner loop fast, the unavoidable thermal contact resistance that exists of chip upper surface, thermal capacitance fluctuation are had stronger robustness; The chip lower surface is the main transport interface of chip heat energy, in its variation that influences the present chip internal fluid temperature of final body to chip, this variation is consistent with the variation tendency of given signal, has objectively shared most tasks of cascade control system outer shroud.The final accurately control of chip temperature is finished by the outer shroud of cascade control system.
Each pcr chip all is subjected to the effect of its below interchanger and top well heater two aspects, and above analytic process thinks that the heat transfer process of all chips in each interchanger isothermal region is identical, has ignored the various dispersivenesses that objectively exist.Dispersiveness with the upper and lower surperficial thermal contact resistance of chip in all polymolecularity has the greatest impact, and the dispersiveness of the upper and lower surperficial thermal contact resistance of chip directly is subjected to the influence of contact pressure dispersiveness.Contact pressure is produced by the mechanical elasticity structure, and its homogeneity is guaranteed by the science design and accurate manufacturing of physical construction.But there is small random fluctuation unavoidably in the contact pressure of each chip in actual use, this small random fluctuation does not cause obvious influence to the polymerase chain reaction process of chip, be within the allowed band, can be considered ground unrest, needn't be comprehended.What need prevent is the obviously inhomogeneous of each chip contact pressure of causing owing to misoperation or plant machinery structure failure, and it will destroy the representativeness of model chip, cause the failure of polymerase chain reaction.Adopt indirect parameter to constitute the supervision and control of each chip contact pressure for this reason.Include the dispersiveness of interface temperature (that is chip upper surface temperature) between each chip of interchanger isothermal region and the flexible printed circuit board plate in statistical Process Control, both can find the irregular variation of the contact pressure that may occur at any time.Statistical Process Control is a kind of important method of production quality control, its basic concept is: quality product is influenced by random factor and abnormal factors two classes, random factor is an outwardness, inevitable, but it is less to the quality product influence, and obeys certain statistical distribution rule.Abnormal factors does not always exist, but bigger to the quality product influence.When abnormal factors did not exist, quality product only was subjected to stochastic factor, and mass parameter is obeyed unified the distribution, is accepting within the scope, claimed that technological process is in the statistics slave mode; And when abnormal factors occurred, huge fluctuation took place in quality product, no longer obeyed original statistical distribution rule, claimed that quality product is not in the statistics slave mode.In case this situation occurs, and should make respective handling immediately, makes quality product return to the statistics slave mode.The effect of statistics control is according to the mathematical statistics theory, and the processing parameter data of continuous acquisition are carried out quantitative statistical study, whether is in the statistics slave mode to judge technological process.When process capability decline occurring, process is out of control or tendency of being out of control arranged, give the alarm immediately, so that in time search, get rid of abnormal factors, make technological process be in the statistics slave mode always.Statistical process control technology has played the effect of the large quantities of unacceptable products of " obviate " appearance.Particular case according to native system, with the difference of each chip upper surface temperature and model chip upper surface temperature as monitoring parameter, select monodrome---the moving range control chart carries out statistical Process Control to above 108 temperature difference parameters altogether, in fact just the dispersiveness of inner each the chip contact pressure of each isothermal region is carried out statistical Process Control, approximately equivalent carries out statistical Process Control in the dispersiveness to the upper and lower surperficial thermal contact resistance of chip.
Be located under the statistics slave mode interface temperature x on j chip of i isothermal region
Ij(i=1,2 ... 12; J=1,2 ... 9) with corresponding isothermal region model chip on interface temperature x
iPoor x
Ij-x
iObey distribution N (μ
Ij, σ
Ij 2).Through Analysis on Mechanism and experimental verification, find μ
Ij≈ 0, σ
Ij≈ σ
2The regularity of distribution that is each temperature difference parameters is approximate consistent, for N (0, σ
2).Select j chip of i isothermal region, extract x
Ij-x
iContinuous n sampled data x
Ijk-x
Ik(k=1,2 ... n), then moving range is defined as:
R
sk＝|(x
ijk-x
ik)-(x
ij(k+1)-x
i(k+1))|，k＝1，2，…n-1 (4)
And average moving range is
Because the regularity of distribution of each chip temperature difference parameters is approximate consistent, therefore adding up the average moving range that obtains from j chip of i isothermal region under the slave mode
Be applicable to all chips.System gathers each temperature data in real time and does above-mentioned computing, but do not need to get ready in real time on the CRT mapping again when work, and monodrome--the moving range control chart lies in the working process of system.Under the situation of control chart data point random alignment, if meet one of following condition, then the decidable system works is under the statistics slave mode, and each chip thermal contact resistance is that the model chip has sufficient representativeness uniformly:
(1) continuous 25 data points, d=0 out-of-bounds counts;
(2) continuous 35 data points, d≤1 of out-of-bounds counting;
(3) continuous 100 data points, d≤2 of out-of-bounds counting;
When one of following situation occurring, can decision-making system be subjected to the influence of abnormal factors, have broken away from the statistics slave mode, the thermal contact resistance of respective chip has obviously departed from normal value, the model chip no longer possesses the representativeness to this chip, and computer system provides corresponding caution:
(1) data point out-of-bounds or pressure circle just declare different;
(2) the some arrangement is not just declared different at random in the boundary.Comprise that idea does periodical change etc. near control limit, idea in the one-sided chaining of control central line, idea repeatedly.
By introducing statistical Process Control, the representativeness of model chip is under the real-time monitoring of system self, though the actual pcr chip self that carries out the polymerase chain reaction does not contain temperature sensor, system can guarantee the temperature control precision to them.
Claims (6)
1, a kind of control device of PCR chip array, it comprises interchanger, place the PCR chip that has at least one microcavity at the interchanger upper surface, it is pcr chip, it is characterized in that: described pcr chip has a plurality of, and they constitute the pcr chip array at the interchanger upper surface; Above the pcr chip array, flexible printed circuit board is set, on flexible printed circuit board, is manufactured with and corresponding well heater of chip array and sensor array; Simultaneously, the interchanger upper surface is divided into some warm areas, one group of pcr chip in the corresponding pcr chip array of each warm area, in each warm area, select a pcr chip as observing and controlling model chip, in model chip microcavity, place micro temperature sensor and be full of the liquid that has approximate thermal characteristic with the PCR reaction solution; Electric components such as temperature sensor, the well heater on the flexible printed circuit board and sensor array in the various kinds board chip are connected with computer control system by interface circuit respectively.
2, the control device of PCR chip array according to claim 1 is characterized in that:
Be shaped on a plurality of unit that constitute by well heater and transmitter on the described flexible printed circuit board, the corresponding pcr chip in each unit, these unit constitute one and corresponding well heater of pcr chip array and sensor array; Flexible printed circuit board is placed in the top of pcr chip array and contacts with each pcr chip.
3, the control device of PCR chip array according to claim 1 is characterized in that:
Is mobile heat exchange medium in the described interchanger, interchanger is the molectron of thermally conductive material and lagging material, and bottom and heat exchange medium gangway are lagging material, and all the other are thermally conductive material, and thermally conductive material forms some grooves and contacts with the heat exchange medium; The interchanger upper surface is divided into several warm areas, with the homogeneity of guaranteeing each warm area upper surface temperature in certain allowed band; Fluid intake place at each warm area is provided with at least one temperature sensor; Corresponding some the pcr chips of the interchanger upper surface of each warm area, wherein one for observing and controlling model chip all the other are the actual chip that carries out the PCR reaction; Interchanger is connected with four liquid-storing boxs, recycle pumps by pipeline, pipeline is provided with eight magnetic valves, be respectively equipped with temperature sensor, heating unit or cooling module in four liquid-storing boxs, these electrical elements all are connected with computer control system by interface circuit, each magnetic valve carries out break-make and switches under computer control, heat exchange medium in the corresponding liquid-storing box is circulated by interchanger, and the heat exchanger surface temperature reaches the numerical value that needs rapidly.
4, a kind of control method of PCR chip array, this method is:
Place at the interchanger upper surface and somely to have microcavity but do not contain the pcr chip of well heater, transmitter, form array; The flexible printed circuit board that will have the array of being made up of well heater and sensor unit is placed on pcr chip array top and contact with it, and each well heater and sensor unit are corresponding one by one with each pcr chip;
Under the effect of computer control system, each pcr chip is by the temperature of interchanger upper surface acquisition near polymerase chain reaction ideal temperature value; Computer control system is gathered each warm area heat exchanger entrance place's heat exchange medium temperature, model chip upper surface temperature, model chip internal fluid temperature, implement the compound control strategy of feedforward one tandem, corresponding well heater on the control flexible printed circuit board, to corresponding pcr chip heating, make it obtain temperature value accurately;
Gather the poor of each chip upper surface temperature and its model chip upper surface temperature simultaneously, implement statistical Process Control, whether be in standard state to monitor each chip thermal contact resistance.
5, the control method of PCR chip array according to claim 4 is characterized in that:
Described feedforward one tandem control mode is: each warm area inlet heat exchange medium temperature detection signal inputs to computer control system in the interchanger, this signal after the given signal generator conversion that inside computer system forms via software on the one hand as the setting signal that is the principal controller that forms by software equally; Mutually comprehensive via the dynamic Feedforward link of software formation on the other hand with the output signal of this principal controller; Detect various kinds board chip upper surface temperature signal simultaneously and form interior ring as the feedback signal of corresponding warm area submaster controller, submaster controller is also formed by software; The temperature of various kinds board chip internal liquid is formed outer shroud as the feedback signal of corresponding warm area principal controller; The submaster controller output signal is through putting on each well heater on the pairing flexible printed circuit board of this group pcr chip after the power magnification simultaneously, to finish each pcr chip accuracy control over temperature.
6, the control method of PCR chip array according to claim 4 is characterized in that:
Described statistical Process Control is, with the difference of chip upper surface temperature and this chip warm area model of living in chip upper surface temperature as monitoring parameter, carry out monodrome-moving range control, promptly the dispersiveness of thermal contact resistance between chip upper surface and the flexible printed circuit board has been carried out statistical Process Control, indirectly the dispersiveness of the upper and lower surperficial contact pressure of chip is carried out statistical Process Control, be equivalent to the dispersiveness of thermal contact resistance between chip lower surface and the interchanger has also been carried out statistical Process Control; When system works in statistics slave mode following time, can think the dispersiveness of each chip thermal contact resistance within allowed band, the model chip has sufficient representativeness; When the influence owing to abnormal factors makes certain chip monitoring parameter break away from the statistics slave mode, think that then the thermal contact resistance of this chip has obviously departed from normal value, the model chip no longer possesses the representativeness to this chip, and computer system provides corresponding caution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100432439A CN100374540C (en) | 2006-03-21 | 2006-03-21 | Controling device and method for polymerase chain reaction chip array |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2006100432439A CN100374540C (en) | 2006-03-21 | 2006-03-21 | Controling device and method for polymerase chain reaction chip array |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1858201A true CN1858201A (en) | 2006-11-08 |
| CN100374540C CN100374540C (en) | 2008-03-12 |
Family
ID=37297159
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006100432439A Expired - Fee Related CN100374540C (en) | 2006-03-21 | 2006-03-21 | Controling device and method for polymerase chain reaction chip array |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100374540C (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102154099A (en) * | 2010-12-20 | 2011-08-17 | 西安交通大学 | Miniature thermal cycle control system for polymerase chain reaction (PCR) biochip |
| CN103777662A (en) * | 2014-01-18 | 2014-05-07 | 浙江大学 | Device and method applied in gradient temperature control |
| CN103781543A (en) * | 2011-08-22 | 2014-05-07 | 松下电器产业株式会社 | Micro fluid device |
| CN108089027A (en) * | 2017-12-28 | 2018-05-29 | 中国电子科技集团公司第十三研究所 | Sensor and navigation attitude instrument based on MEMS capacitive micro-acceleration gauge |
| WO2019144905A1 (en) * | 2018-01-24 | 2019-08-01 | 北京光阱管理咨询合伙企业（有限合伙） | Microdroplet container, method for preparing microdroplet container, microdroplet spreading method, microdroplet formation kit, temperature control device, oil phase composition for microdroplet formation and treatment method therefor |
| CN115420307A (en) * | 2022-11-04 | 2022-12-02 | 四川凌翔科技发展有限公司 | Real-time self-calibration self-diagnosis low-temperature drift data acquisition system |
| US11666900B2 (en) | 2018-01-24 | 2023-06-06 | Sniper (Suzhou) Life Technology Co. | Motion controlling mechanism, liquid discharging nozzle, microdroplet generating device and method, liquid driving mechanism and method, microdroplet generating method, and surface processing method of liquid discharging nozzle |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6261431B1 (en) * | 1998-12-28 | 2001-07-17 | Affymetrix, Inc. | Process for microfabrication of an integrated PCR-CE device and products produced by the same |
| CN2612673Y (en) * | 2003-03-27 | 2004-04-21 | 上海复生生物工程研究所有限公司 | Full-automatic water-bath type gene amplification |
-
2006
- 2006-03-21 CN CNB2006100432439A patent/CN100374540C/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102154099A (en) * | 2010-12-20 | 2011-08-17 | 西安交通大学 | Miniature thermal cycle control system for polymerase chain reaction (PCR) biochip |
| CN103781543A (en) * | 2011-08-22 | 2014-05-07 | 松下电器产业株式会社 | Micro fluid device |
| CN103777662A (en) * | 2014-01-18 | 2014-05-07 | 浙江大学 | Device and method applied in gradient temperature control |
| CN103777662B (en) * | 2014-01-18 | 2016-01-13 | 浙江大学 | A kind of apparatus and method being applied to gradient temperature and controlling |
| CN108089027A (en) * | 2017-12-28 | 2018-05-29 | 中国电子科技集团公司第十三研究所 | Sensor and navigation attitude instrument based on MEMS capacitive micro-acceleration gauge |
| WO2019144905A1 (en) * | 2018-01-24 | 2019-08-01 | 北京光阱管理咨询合伙企业（有限合伙） | Microdroplet container, method for preparing microdroplet container, microdroplet spreading method, microdroplet formation kit, temperature control device, oil phase composition for microdroplet formation and treatment method therefor |
| US11666900B2 (en) | 2018-01-24 | 2023-06-06 | Sniper (Suzhou) Life Technology Co. | Motion controlling mechanism, liquid discharging nozzle, microdroplet generating device and method, liquid driving mechanism and method, microdroplet generating method, and surface processing method of liquid discharging nozzle |
| US11946100B2 (en) | 2018-01-24 | 2024-04-02 | Sniper (Suzhou) Life Technology Co., Ltd. | Microdroplet container and method for manufacturing the same, method for spreading microdroplets, microdroplet-generating kit, temperature-controlling device, oil phase composition for microdroplet generating and method for treating the same |
| CN115420307A (en) * | 2022-11-04 | 2022-12-02 | 四川凌翔科技发展有限公司 | Real-time self-calibration self-diagnosis low-temperature drift data acquisition system |
| CN115420307B (en) * | 2022-11-04 | 2023-02-07 | 四川凌翔科技发展有限公司 | Real-time self-calibration self-diagnosis low-temperature drift data acquisition system |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100374540C (en) | 2008-03-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1858201A (en) | Controling device and method for polymerase chain reaction chip array | |
| CN102046291B (en) | Thermal control system and method for chemical and biochemical reactions | |
| CN103003448B (en) | Improved thermal uniformity for thermal cycler instrumentation using dynamic control | |
| CN103034169B (en) | Modeling and compensation method of heat error of numerical control machine tool | |
| CN101281121A (en) | Quick changeable temperature small-sized constant temperature sample pool | |
| US11684916B2 (en) | Chip for polymerase chain reaction, method of operation chip, and reaction device | |
| CN108195875B (en) | System and method for rapidly and automatically measuring cold and hot circulation of phase change material in wide temperature area | |
| CN104703471A (en) | Method and device for thawing biological material | |
| CN102601987A (en) | Temperature control method and system of air-pressure thermoforming machine | |
| CN204883432U (en) | Reaction chamber temperature controlling means , temperature control system and hematology analyzer | |
| US20220401960A1 (en) | High throughput radiochemistry system | |
| CN103677011B (en) | One is applicable to extend blackbody wide temperature range control system under vacuum condition | |
| CN106442454A (en) | Quick gene amplification detection device and quick gene amplification detection method based on fluorescent quantitation | |
| CN109471037A (en) | A kind of salt bridge-type thermoelectrochemistry battery testing platform quickly tested and its method | |
| WO2015003722A1 (en) | Single-use device with a reaction chamber and a method for controlling the temperature in the device and uses thereof | |
| CN103415346B (en) | Control system and method for biologic applications | |
| CN102636517B (en) | Device and method for testing pressure-volume-temperature (PVT) relationship of polymer at high cooling rate | |
| CN113155601A (en) | Array film force and thermal parameter test loading system and loading method | |
| US20220105483A1 (en) | Method and device for controlling the temperature of reaction mixtures in an agitation operation | |
| CN201575992U (en) | Small-sized thermostat | |
| CN106876824B (en) | A kind of semiconductor chilling plate Double-direction Temperature control device for button cell | |
| CN202494940U (en) | Temperature control system of air pressure thermoforming machine | |
| CN206975459U (en) | Two-component metal phase diagram experimental provision based on Single chip microcomputer temperature control system | |
| CN105005336A (en) | Temperature control device for viscoelastic damper performance test | |
| CN108499153A (en) | A kind of parallel crystallization instrument temperature control equipment of high throughput and method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080312 Termination date: 20110321 |