CN1584043A - Spiral microchannel polymerase chain reacting chip - Google Patents
Spiral microchannel polymerase chain reacting chip Download PDFInfo
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- CN1584043A CN1584043A CN 200410024703 CN200410024703A CN1584043A CN 1584043 A CN1584043 A CN 1584043A CN 200410024703 CN200410024703 CN 200410024703 CN 200410024703 A CN200410024703 A CN 200410024703A CN 1584043 A CN1584043 A CN 1584043A
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Abstract
A spiral microchannel polymerase chain reacting chip for preventive medicine and hygiene is disclosed. Basal layer with microflow-channel sealing with sealing cover coat by bonding technology, reagent sample inlet and outlet in cover coat, corresponding to channel origin and terminal, circular denaturating zone heater, circular extending zone heater and circular temperature sensor on another side of basal layer of microflow-channel, renaturating zone radiating zone with metal thin-membrane formation between insulating layer at right bottom of chip and basal layer with microflow-channel, insulating layer and metal wiring layer depositing on basal layer with microflow-channel. It can be used to improve compatibility, integration and efficiency of chip.
Description
Technical field
What the present invention relates to is a kind of PCR chip, particularly a kind of spiral microchannel PCR chip.Be used for preventive medicine and health field.
Background technology
In molecular biological chemical analysis system, the polymerase chain reaction is vital composition link, and the development of micro-electromechanical technology (MEMS) can be finished the polymerase chain reaction on chip.General micropassage type PCR chip, control sex change, renaturation, three reactions steps of extension respectively with three humidity provinces, be made in the variation repeatedly of these three humidity provinces of microchannel circulation process on the chip, make in the mix reagent of participating in reaction in the passage, the fragment of thymus nucleic acid (DNA) molecule obtains the amplification of geometricprogression level when flowing in microchannel.The micropassage type PCR chip is the focus of current PCR chip research, result of study show the micropassage type PCR chip with the characteristics of himself structure overcome the heating of microchamber static reaction chip slowly, be difficult to use in the difficulty of sample introduction and crossed contamination in batches, efficient in the molecular chemistry analysis is improved greatly, can be widely used in fields such as biomedicine, preventive medicine and environmental monitoring.
The efficient of polymerase chain reaction and reagent are in sex change, renaturation, the temperature and time that extends three links is closely related, the degree that the circulating temperature change procedure of reagent experience and polymerase chain reaction temperature curve coincide influences the formation efficiency of product to a great extent, but, current micropassage type PCR chip is unreasonable all the time in the design of microfluidic circuit, in the design of majority, the capital makes microfluidic channel after through three humidity provinces in the return course, the waiting period of inevitably through one section centre, it not only makes the micro-thymus nucleic acid dna molecular fragment in the reagent experience sex change, renaturation, can't completely meet the polymerase chain reaction temperature curve when extending three temperature variation, also reduced the speed of polymerase chain reaction, be unfavorable for amplification efficiency.Find by literature search, people such as Ivonne Schneega β are in " Reviews in MolecularBiotechnology " (" molecular biology comment ") 2001, " Flow-throughpolymerase chain reactions in chip thermocyclers " (the micropassage type PCR chip) delivered on the 82:101-121, the PCR chip TC1997 that mentions in this article has 25 circulation passages, the design of the sectional area of fluid channel waiting period of adopting in the middle of reducing, the inner fluid passage flow velocity is improved, yet, owing to change miniflow pipeline sectional area, can produce the bubble that hinders the polymerase chain reaction in the microchannel, in addition, because the ununiformity of its flow velocity, this micropassage type PCR chip can't be applied to the needs of sample introduction in batches, in fact, this design can't be used for practical application.Also mentioned the micropassage type PCR chip of several structures in the literary composition, though improved the layout of hot-zone, utilize special process and material to improve amplification efficiency, but on the arrangement space of fluid channel and three humidity provinces, all there is identical shortcoming, is unfavorable for high-level efficiency polymerase chain reaction needs with TC1997.
Summary of the invention
The objective of the invention is to the not enough and irrational defective of microchannel layout of humidity province design at existing PCR chip, a kind of spiral microchannel PCR chip is proposed, by changing microfluidic circuit layout and humidity province layout on the sheet, when reagent is flowed on chip, the temperature variation of experience meets the amplification curve of polymerase chain reaction, thereby improve the productive rate of amplification, solved the problem that exists in the background technology.
The present invention is achieved through the following technical solutions.Chip of the present invention comprises: the sealing cap rock, the stratum basale, reagent sample delivery port, the reagent sample input aperture that have microchannel, spiral microchannel, insulation layer and metal lead wire floor, annular temperature sensor, renaturation area radiating area and annular sex change district well heater, annular extension area well heater.Its mode of connection is: the microchannel that has on the stratum basale of microchannel is spiral microchannel, the stratum basale that has microchannel is sealed with bonding techniques with the sealing cap rock, be provided with the reagent sample input aperture at the sealing cap rock, the reagent sample delivery port, the reagent sample input aperture, the reagent sample delivery port respectively with the passage starting point, channel end is corresponding, the opposite side that has the stratum basale of microchannel is provided with annular sex change district well heater, annular extension area well heater and annular temperature sensor, the film formed renaturation area radiating area of metal foil is positioned at the chip lower right in addition, at the insulation layer of chip with have between the stratum basale of microchannel, be responsible for when renaturation area humidity province too high, participating in forced heat radiation, have depositing insulating layer and metal lead wire layer on the stratum basale of microchannel.
Spiral microfluidic channel, in on layout, dividing the endless belt microfluidic channel with outside the endless belt microfluidic channel, interior endless belt microfluidic channel is in the chip center zone, the inactivation of ThermoScript II when responsible reagent preheats with inverse transcription polymerase chain reaction, outer endless belt microfluidic channel is corresponding with each temperature heater that is positioned at chip bottom, is divided into sex change district, renaturation area, extension area.
The basic structure that has the stratum basale and the sealing cap rock compositing chip of microchannel, be provided with in chip bottom: reagent preheats district's well heater, sex change district annular membrane well heater, renaturation area radiating area, extension area annular membrane well heater, when reagent sample is moved on spiral microchannel, stable flowing through preheats district, sex change district, renaturation area, extension area, its temperature is respectively distinguished annular membrane well heater and annular temperature sensor monitors by what be positioned at the chip bottom, and multiplication product sample is derived by the reagent sample delivery port.For level and smooth reagent sample flows, spiral microchannel has designed fillet at the joining place of flex point section.
Among the present invention, in suprabasil spiral microfluidic channel is divided on layout endless belt and outside endless belt, the effect of interior endless belt microfluidic channel is: when reagent flows in interior endless belt microfluidic channel, preheat the effect of district's well heater through reagent, acquisition is fully unwind before entering circulation, improve productive rate, and for the special polymerase chain reaction-inverse transcription polymerase chain reaction of another kind, in reaction process, the ThermoScript II of sneaking in the liquid reagent sample can suppress the polymerase chain reaction, when reagent was mobile from interior endless belt microfluidic channel, the heat-processed of utilizing reagent to preheat the district can make the ThermoScript II inactivation, makes it be unlikely to enter the amplified reaction link.And outer endless belt microfluidic channel is made of sex change district, renaturation area, extension area, when reagent flows through when respectively distinguishing successively, be positioned at the integrated annular membrane well heater of respectively distinguishing of the chip bottom heating and cooling reagent that can circulate, the cycle of polymerase chain reaction process is carried out.
Promptly interior endless belt microfluidic channel bottom, the central zone of spiral microchannel is provided with and preheats district's well heater, and with outer endless belt microfluidic channel sex change district, extension area is corresponding is annular sex change district well heater, annular extension area well heater and annular temperature sensor, the joining place of each flex point section of spiral microchannel adopts the level and smooth reagent sample of fillet to flow, and preheats district's well heater and annular sex change district well heater, annular extension area well heater and is annular.
The winding pipeline changing that declines that the present invention will have delivery reagent sample in the PCR chip now becomes spiral microchannel.Reagent is preheating, the sex change district, renaturation area, the heat-up time of extension area is by the hot-zone area, flow rate of fluid and the microchannel shape on sheet decides, spiral microchannel is determined by serial involute urve parameter in suprabasil shape, the temperature regulation of each hot-zone realizes by being positioned at integrated annular membrane well heater of respectively distinguishing of chip bottom and loop sensor, the interior ring girth of spiral microchannel is less than outer shroud, after the liquid reagent sample injects from the reagent sample input aperture, endless belt microfluidic channel in being introduced into, begin the amplification process, rising along with target sequence concentration in the reagent, reagent begins to enter outer endless belt microfluidic channel, also prolong the heat-up time in three humidity provinces of being responsible for the polymerase chain reaction thereupon, the multiplication after product is flowed out by the reagent sample delivery port that is positioned at spiral microchannel outer shroud end.The annular membrane well heater of chip bottom and loop sensor are accurately controlled the carrying out that guarantees polymerase chain reaction on the chip by computer.The present invention can further reduce chip volume, thereby makes its easier being integrated in the chipset bulk analysis system that has the comprehensive analysis processing ability.
The present invention makes in package shape with glass and two kinds of materials of polydimethylsiloxane (PDMS), utilize soft lithographic technology on PDMS, to produce substrate with spiral microfluidic channel, with make it airtight after the glass capping encapsulation, reagent sample input aperture in the glass capping, reagent sample delivery port are responsible for the input of reagent and the output of product, under the substrate of microchannel layers be the chip bottom integrated annular membrane well heater and loop sensor controlled temperature, thin film heater and transmitter adopt sputtered platinum to belong to (Pt) or indium tin oxide (ITO) film is realized.On integrated annular membrane well heater and annular temperature sensor, form insulation layer again, last cloth lead-in wire and electrode.
The present invention has substantial characteristics and progress, the present invention has changed the fluid channel layout of existing microchannel PCR chip, the distribution of hot-zone is more become rationally, and sample reagent gets the more approaching amplification temperature curve of temperature cycle, thereby has improved amplification efficiency.Preheat fluid channel, district and chip bottom and interior ring passage respective films well heater and transmitter by ring in designing, enlarged the use range of this chip, strengthen the efficient of chip, dwindled chip volume simultaneously, but improved the compatibility and the integration of chip.
Description of drawings
Fig. 1 structural representation of the present invention
Fig. 2 left view decomposing schematic representation of the present invention
Fig. 3 right view decomposing schematic representation of the present invention
The spiral microfluidic circuit channel architecture of Fig. 4 the present invention synoptic diagram
Fig. 5 annular membrane heater structure of the present invention synoptic diagram
Embodiment
As Fig. 1,2,3,4, shown in 5, the present invention includes: sealing cap rock 1, the stratum basale 2 that has microchannel, spiral microchannel 4, reagent sample delivery port 6, reagent sample input aperture 7, insulation layer 12 and metal lead wire layer 13, annular temperature sensor 18, renaturation area radiating area 19 and annular sex change district well heater 21, annular extension area well heater 22, its mode of connection is: the microchannel that has on the stratum basale 2 of microchannel is spiral microchannel 4, the stratum basale 2 that has microchannel is packaged as a whole with sealing cap rock 1, be provided with reagent sample input aperture 7 at sealing cap rock 1, reagent sample delivery port 6, reagent sample input aperture 7, reagent sample delivery port 6 respectively with passage starting point 14, channel end 16 is corresponding, the opposite side that has the stratum basale 2 of microchannel is provided with annular sex change district well heater 21, annular extension area well heater 22 and annular temperature sensor 18, the film formed renaturation area radiating area 19 of metal foil is positioned at the chip lower right in addition, as Fig. 3, at the insulation layer 12 of chip with have between the stratum basale 2 of microchannel, be responsible for when renaturation area humidity province too high, participating in forced heat radiation, have depositing insulating layer 12 and metal lead wire layer 13 on the stratum basale 2 of microchannel.
As Fig. 2,4, spiral microfluidic channel 4, in on layout, dividing the endless belt microfluidic channel with outside the endless belt microfluidic channel, interior endless belt microfluidic channel is in the chip center zone, the inactivation of ThermoScript II when responsible reagent preheats with inverse transcription polymerase chain reaction, outer endless belt microfluidic channel is corresponding with each temperature heater that is positioned at chip bottom, is divided into sex change district, renaturation area, extension area.
The central zone of spiral microchannel 4 promptly in endless belt microfluidic channel bottom be provided with and preheat district's well heater 20, and with outer endless belt microfluidic channel sex change district, extension area is corresponding is annular sex change district well heater 21, annular extension area well heater 22 and annular temperature sensor 18.Preheat district's well heater 20 and annular sex change district well heater 21, annular extension area well heater 22 and be annular.The joining place of the flex point section of each section of spiral microchannel 4 adopts fillet 5,15,17 designs to flow with level and smooth reagent sample.
The present invention has adopted the multilayered structure pattern, have the stratum basale 2 of microchannel and the basic structure that sealing cap rock 1 forms chip, the chip bottom is: reagent preheats the annular membrane well heater 9 of district's well heater 10 and control sex change district temperature, the thin film heater 11 of control renaturation area heat radiation, the thin film heater 8 of control extension area temperature, when reagent sample is moved on spiral microchannel 4, flow through sex change, renaturation, extension area successively, be positioned at the respective heater and the transmitter controlled temperature of chip bottom, final multiplication product sample is derived by reagent sample delivery port 6.
Claims (5)
1, a kind of spiral microchannel PCR chip, comprise: sealing cap rock (1), the stratum basale (2) that has microchannel, spiral microchannel (4), reagent sample delivery port (6), reagent sample input aperture (7), insulation layer (12) and metal lead wire layer (13), annular temperature sensor (18), renaturation area radiating area (19) and annular sex change district well heater (21), annular extension area well heater (22), it is characterized in that, the microchannel that has on the stratum basale (2) of microchannel is spiral microchannel (4), the stratum basale (2) that has microchannel is packaged as a whole with sealing cap rock (1), be provided with reagent sample input aperture (7) at sealing cap rock (1), reagent sample delivery port (6), reagent sample input aperture (7), reagent sample delivery port (6) respectively with passage starting point (14), channel end (16) is corresponding, the opposite side that has the stratum basale (2) of microchannel is provided with annular sex change district well heater (21), annular extension area well heater (22) and annular temperature sensor (18), the film formed renaturation area radiating area of metal foil (19) is positioned at chip lower right insulation layer (12) and has between the stratum basale (2) of microchannel in addition, and the stratum basale (2) that has microchannel is gone up depositing insulating layer (12) and metal lead wire layer (13).
2, spiral microchannel according to claim 1 PCR chip, it is characterized in that, adopted the multilayered structure pattern, have the stratum basale (2) of microchannel and the basic structure that sealing cap rock (1) forms chip, the chip bottom is: reagent preheats the annular membrane well heater (9) of district's well heater (10) and control sex change district temperature, the thin film heater (11) of control renaturation area heat radiation, the thin film heater (8) of control extension area temperature.
3, spiral microchannel according to claim 1 PCR chip, it is characterized in that, described spiral microfluidic channel (4), in on layout, dividing the endless belt microfluidic channel with outside the endless belt microfluidic channel, interior endless belt microfluidic channel is in the chip center zone, the inactivation of ThermoScript II when responsible reagent preheats with inverse transcription polymerase chain reaction, outer endless belt microfluidic channel is corresponding with each temperature heater that is positioned at chip bottom, is divided into sex change district, renaturation area, extension area.
4, according to claim 1 or 3 described spiral microchannel PCR chips, it is characterized in that, the interior endless belt microfluidic channel bottom of described spiral microchannel (4) is provided with and preheats district's well heater (20), with outer endless belt microfluidic channel sex change district, extension area is corresponding is annular sex change district well heater (21), annular extension area well heater (22) and annular temperature sensor (18), preheat district's well heater (20) and annular sex change district well heater (21), annular extension area well heater (22) and be annular.
According to claim 1 or 3 described spiral microchannel PCR chips, it is characterized in that 5, the joining place of each flex point section of described spiral microchannel (4) adopts the level and smooth reagent sample of fillet (5,15,17) to flow.
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| CN 200410024703 CN1242072C (en) | 2004-05-27 | 2004-05-27 | Spiral microchannel polymerase chain reacting chip |
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| CN 200410024703 CN1242072C (en) | 2004-05-27 | 2004-05-27 | Spiral microchannel polymerase chain reacting chip |
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Cited By (11)
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| CN1319848C (en) * | 2005-09-22 | 2007-06-06 | 上海交通大学 | Array type piezoelectricity-driven multi-cavity micro-mixer |
| CN100385227C (en) * | 2005-09-08 | 2008-04-30 | 上海交通大学 | Piezo-electric driving type micro mixed chip |
| CN101187609B (en) * | 2005-09-08 | 2010-06-30 | 上海交通大学 | Piezo-electric drive type micro mixing chip |
| CN101851680A (en) * | 2010-06-01 | 2010-10-06 | 厦门大学 | Method for biochip high-throughput hybridization |
| CN102071136A (en) * | 2009-11-25 | 2011-05-25 | 中国科学院电子学研究所 | Array micro polymerase chain reaction chip |
| CN102220226A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Two-path temperature control polymerase chain reactor and real-time detection device |
| CN102220225A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Polymerase chain reactor and real-time electromechanical scanning and detecting device |
| CN102220227A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Polymerase chained type reactor and real-time micro-optics detection device |
| CN102220228A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Polymerase chain reactor and real-time optical array detection device |
| CN102713563A (en) * | 2009-10-06 | 2012-10-03 | 雷根斯堡学院 | Miniaturised online trace analysis |
| JP2018089623A (en) * | 2009-10-15 | 2018-06-14 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Digital microfluidic platform for radiochemistry |
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- 2004-05-27 CN CN 200410024703 patent/CN1242072C/en not_active Expired - Fee Related
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100385227C (en) * | 2005-09-08 | 2008-04-30 | 上海交通大学 | Piezo-electric driving type micro mixed chip |
| CN101187609B (en) * | 2005-09-08 | 2010-06-30 | 上海交通大学 | Piezo-electric drive type micro mixing chip |
| CN1319848C (en) * | 2005-09-22 | 2007-06-06 | 上海交通大学 | Array type piezoelectricity-driven multi-cavity micro-mixer |
| CN102713563A (en) * | 2009-10-06 | 2012-10-03 | 雷根斯堡学院 | Miniaturised online trace analysis |
| CN102713563B (en) * | 2009-10-06 | 2015-09-23 | 雷根斯堡学院 | The online trace analysis of miniaturization |
| JP2018089623A (en) * | 2009-10-15 | 2018-06-14 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Digital microfluidic platform for radiochemistry |
| CN102071136A (en) * | 2009-11-25 | 2011-05-25 | 中国科学院电子学研究所 | Array micro polymerase chain reaction chip |
| CN101851680A (en) * | 2010-06-01 | 2010-10-06 | 厦门大学 | Method for biochip high-throughput hybridization |
| CN102220228A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Polymerase chain reactor and real-time optical array detection device |
| CN102220227A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Polymerase chained type reactor and real-time micro-optics detection device |
| CN102220226B (en) * | 2011-05-23 | 2013-07-24 | 北京工业大学 | Two-path temperature control polymerase chain reactor and real-time detection device |
| CN102220225A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Polymerase chain reactor and real-time electromechanical scanning and detecting device |
| CN102220226A (en) * | 2011-05-23 | 2011-10-19 | 北京工业大学 | Two-path temperature control polymerase chain reactor and real-time detection device |
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| CN1242072C (en) | 2006-02-15 |
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