CN113862144A - Full-automatic fluorescent quantitative PCR analyzer - Google Patents
Full-automatic fluorescent quantitative PCR analyzer Download PDFInfo
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- CN113862144A CN113862144A CN202111307008.9A CN202111307008A CN113862144A CN 113862144 A CN113862144 A CN 113862144A CN 202111307008 A CN202111307008 A CN 202111307008A CN 113862144 A CN113862144 A CN 113862144A
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- 238000003753 real-time PCR Methods 0.000 title claims abstract description 25
- 238000006243 chemical reaction Methods 0.000 claims abstract description 103
- 238000001514 detection method Methods 0.000 claims abstract description 81
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 76
- 230000005284 excitation Effects 0.000 claims abstract description 37
- 238000010438 heat treatment Methods 0.000 claims abstract description 36
- 230000000630 rising effect Effects 0.000 claims abstract description 7
- 239000013307 optical fiber Substances 0.000 claims description 75
- 230000003287 optical effect Effects 0.000 claims description 26
- 230000007246 mechanism Effects 0.000 claims description 19
- 238000001917 fluorescence detection Methods 0.000 claims description 16
- 230000000712 assembly Effects 0.000 claims description 10
- 238000000429 assembly Methods 0.000 claims description 10
- 238000009434 installation Methods 0.000 claims description 9
- 238000009413 insulation Methods 0.000 claims description 8
- 229920000742 Cotton Polymers 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 3
- 230000008878 coupling Effects 0.000 claims description 3
- 238000010168 coupling process Methods 0.000 claims description 3
- 238000005859 coupling reaction Methods 0.000 claims description 3
- 229910021389 graphene Inorganic materials 0.000 claims description 3
- 239000004065 semiconductor Substances 0.000 claims description 3
- 238000004458 analytical method Methods 0.000 claims description 2
- 230000017525 heat dissipation Effects 0.000 claims description 2
- 239000000835 fiber Substances 0.000 abstract description 25
- 238000001816 cooling Methods 0.000 description 9
- 238000010586 diagram Methods 0.000 description 7
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- 238000000034 method Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 3
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- 230000000694 effects Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 238000012921 fluorescence analysis Methods 0.000 description 2
- 229910052736 halogen Inorganic materials 0.000 description 2
- 150000002367 halogens Chemical class 0.000 description 2
- 238000011897 real-time detection Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 1
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- 230000005281 excited state Effects 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
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- 238000004445 quantitative analysis Methods 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1894—Cooling means; Cryo cooling
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- General Health & Medical Sciences (AREA)
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- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a full-automatic fluorescent quantitative PCR analyzer, which comprises a temperature control system and a fluorescent detection module, wherein the temperature control system comprises a temperature rising and reducing module and a hot cover component. The full-automatic fluorescent quantitative PCR analyzer can directly heat the reaction seat and the reagent in the reaction seat by utilizing the heating device arranged on the hot cover; meanwhile, the temperature control assembly is arranged on the bottom surface of the reagent tube seat, and can quickly control temperature rise and fall circulation, namely the temperature rise and fall circulation control of PCR can be realized; through setting up a plurality of interface groups, each interface group corresponds a reaction seat, set up optic fibre between two connectors with the excitation fiber interface of the interface group that corresponds respectively and the receiving fiber interface with the reaction seat and link to each other, each reagent sample all can detect through all determine module, each determine module all can detect all reagent samples, realize that all reagents all can carry out the technical purpose that the multichannel detected, can effectively improve detection efficiency.
Description
Technical Field
The invention belongs to the technical field of medical instruments, and particularly relates to a full-automatic fluorescent quantitative PCR analyzer.
Background
The fluorescence analysis method is a method for performing qualitative or quantitative analysis by utilizing fluorescence which is generated by de-excitation process of collision and emission of excited molecules and can reflect the characteristics of certain substances when the substances are in an excited state after being irradiated by ultraviolet light. Since some substances do not emit fluorescence by themselves (or fluoresce very weakly), it is necessary to convert the non-fluorescent substance into a substance that emits fluorescence. For example, with certain reagents (e.g., fluorescent dyes) that form complexes with non-fluorescent materials, each complex being capable of emitting fluorescence, and the assay is performed. Therefore, the use of the fluorescent reagent opens the door to the fluorescence analysis of some inorganic substances and organic substances which do not originally fluoresce, and expands the analysis range.
The fluorescence quantitative analyzer is a reaction instrument for real-time detection, and the functions of the instrument are generally ensured by a heat circulation system and a fluorescence real-time detection system. The fluorescence quantitative analyzer mainly comprises a temperature control module, a fluorescence detection module, a circuit control module, a computer and processing software thereof. The existing temperature control module realizes the temperature cycle of PCR through air bath, water bath, metal bath and other forms, and although the temperature control module can meet the requirement of temperature rise and fall cycle to a certain extent, the temperature control module has the defects of large volume and low temperature rise and fall speed.
At present, most of fluorescence detection modules of fluorescence quantitative analyzers take pictures or detect reagents through optical fibers during detection. When a photographing mode is adopted, the light source generally adopts a halogen lamp, the halogen lamp generally attenuates after long-term use to cause inaccurate measuring results, and the photographed pictures have edge effects to cause inaccurate measuring results; when the optical fiber mode is adopted, the PMT photomultiplier is required to be matched for detection, and the PMT photomultiplier is expensive and has a large volume, so that the PMT photomultiplier can only be matched with a linear guide rail or a screw rod mechanism to carry out one-by-one scanning mode on the reagent for detection. If the reagent needs to be subjected to multi-channel detection, the channel needs to be replaced and the detection can be switched to the next channel for detection after the detection of the previous channel is completed, so that the detection rate is relatively slow, and if the reagent has a high requirement on the detection rate, the detection mode is not applicable.
Disclosure of Invention
In view of this, the present invention provides a full-automatic fluorescence quantitative PCR analyzer, which can realize PCR temperature rise and fall cycle, prevent excessive evaporation of reagents, and can synchronously realize multi-channel detection, thereby improving detection efficiency.
In order to achieve the purpose, the invention provides the following technical scheme:
a full-automatic fluorescence quantitative PCR analyzer comprises a temperature control system and a fluorescence detection module, wherein the temperature control system comprises a temperature rising and reducing module and a hot cover assembly;
the temperature rising and lowering module comprises a bottom plate and a reagent tube seat assembly; the reagent tube seat assembly comprises a reagent tube seat, the top surface of the reagent tube seat is provided with a reaction seat, a reaction cavity is arranged in the reaction seat, and the bottom surface of the reagent tube seat is provided with a temperature control assembly for controlling the temperature rise and fall circulation in the reaction cavity; the bottom plate is provided with a pressing assembly for pressing and fixing the reagent tube seat;
the hot cover assembly comprises a fixed plate and a hot cover capable of covering the fixed plate; a reaction zone is arranged on the fixing plate, and a reaction through hole for the reaction seat to pass through is arranged in the reaction zone; a heating device is arranged on the hot cover corresponding to the reaction zone; the fixed plate is positioned above the bottom plate, and the fixed plate and the bottom plate are relatively and fixedly arranged; the reaction through holes and the reaction seats are arranged in a one-to-one correspondence manner, and the reaction seats are positioned in the corresponding reaction through holes;
the fluorescence detection module comprises an interface disc, a detection disc and a driving mechanism, wherein a plurality of interface groups are arranged on the interface disc, and each interface group comprises an excitation optical fiber interface and a receiving optical fiber interface; the detection disc is provided with a plurality of detection components, and each detection component comprises an exciting light component and a receiving light component; the driving mechanism drives the interface disc and the detection disc to move relatively, so that each interface group sequentially passes through all the detection assemblies, or each detection assembly sequentially passes through all the interface groups;
the outer side wall of the reaction seat is provided with an optical fiber connecting seat, the optical fiber connecting seat is provided with two connecting ports, and the connecting ports are communicated with the reaction cavity; the interface group is arranged corresponding to the reaction seat, and two connectors of the reaction seat are respectively connected with the excitation optical fiber interface and the receiving optical fiber interface of the corresponding interface group through optical fibers.
Furthermore, step surfaces are respectively arranged on two sides of the reagent tube seat, the pressing assembly comprises tube seat pressing plates respectively positioned on two sides of the reagent tube seat, and the tube seat pressing plates are fixedly connected with the bottom plate and pressed on the corresponding step surfaces;
the bottom plate is provided with an optical fiber fixing plate, the optical fiber fixing plate is provided with a fixing plate limiting groove for limiting an optical fiber, and the optical fiber fixing plate is provided with an optical fiber pressing plate for tightly pressing and fixing the optical fiber in the fixing plate limiting groove;
and a pressing plate limiting groove for limiting the optical fiber is arranged on the tube seat pressing plate between the optical fiber fixing plate and the reagent tube seat, the pressing plate limiting groove corresponds to the fixing plate limiting groove, and the corresponding pressing plate limiting groove and the fixing plate limiting groove are displaced on the same straight line.
Furthermore, a plurality of reaction seats are arranged on the top surface of the reagent tube seat at intervals, and the reaction seats are arranged on the top surface of the reagent tube seat in a one-dimensional linear array; and a thermistor is arranged in the reagent tube seat and is positioned between two adjacent reaction seats.
Further, the temperature rising and lowering module further comprises an over-temperature protection switch for preventing the reagent tube seat from being overhigh in temperature.
Furthermore, the temperature control assembly comprises a temperature control element for controlling temperature rise and fall, and a first heat conduction layer and a second heat conduction layer are respectively arranged on the upper side surface and the lower side surface of the temperature control element; the temperature control element adopts a semiconductor refrigerator; the first heat conduction layer and the second heat conduction layer are both made of graphene.
Furthermore, temperature control system still includes radiator unit, radiator unit includes middle wind channel, the both sides in middle wind channel are equipped with respectively rather than the side wind channel that is linked together, two be equipped with respectively on the side wind channel rise or fall warm mould piece and hot lid subassembly, the equal fixed mounting of fixed plate and bottom plate is in correspondence on the side wind channel.
Furthermore, heat insulation plates used for being in contact fit with the heating device are arranged on the periphery of the reaction area; the top surface of the reaction seat is positioned between the top surface of the reaction zone and the top surface of the heat insulation plate;
the hot cover is provided with a heating installation groove for installing a heating device, the heating device is installed in the heating installation groove, and a flexible elastic element is arranged between the heating device and the bottom of the heating installation groove.
Furthermore, the bottom surface of the fixing plate, which faces away from the hot cover, is provided with heat-insulating cotton, and the heat-insulating cotton is provided with heat-insulating through holes which are used for the reaction seats to pass through in a one-to-one correspondence manner with the reaction through holes.
Furthermore, the connecting end of the thermal cover is hinged with the fixing plate, and a lock catch is arranged between the opening and closing end of the thermal cover opposite to the connecting end and the fixing plate; and an electromagnet is arranged at the opening and closing end of the hot cover and/or the end of the fixing plate provided with the lock catch.
Furthermore, the centers of all the excitation optical fiber interfaces are located on the same circular arc line I, the centers of all the receiving optical fiber interfaces are located on a circular arc line II, and the circle centers of the circular arc line I and the circular arc line II are overlapped; in the axial view direction of the excitation optical fiber interface or the receiving optical fiber interface, the geometric centers of all the excitation optical components are located on the circular arc line I, and the geometric centers of all the receiving optical components are located on the circular arc line II; the driving mechanism is a rotation driving mechanism which drives the interface disc and the detection disc to rotate relatively.
Furthermore, the rotation driving mechanism comprises a stepping motor which is used for driving the detection disc and the interface disc to rotate relatively, and the axis of the relative rotation between the detection disc and the interface disc passes through the circle centers of the arc line I and the arc line II.
Further, the curvature radius of the circular arc line I is larger than that of the circular arc line II; and the included angle between the connecting line between the geometric centers of the exciting light assembly and the receiving light assembly which belong to the same detection assembly and the circle center of the circular arc line I is 30 degrees.
Further, the optical coupling sensor is used for positioning the relative position between the detection disc and the interface disc; and the detection plate is provided with a conductive slip ring.
Further, the exciting light assembly comprises an LED light source, a first convex lens, a second convex lens and a third convex lens which are sequentially arranged, a first optical filter is arranged between the first convex lens and the second convex lens, and the third convex lens is positioned on one side of the second convex lens facing the interface disc;
the receiving optical assembly comprises a fourth convex lens, a fifth convex lens and a PD sensor which are sequentially arranged, a second optical filter is arranged between the fourth convex lens and the fifth convex lens, and the fourth convex lens is positioned on one side of the fifth convex lens facing the interface disc.
The invention has the beneficial effects that:
according to the full-automatic fluorescent quantitative PCR analyzer, the temperature raising and lowering module and the heat cover component are arranged in the temperature control system, the reaction seat of the temperature raising and lowering module is positioned in the corresponding reaction through hole, after the heat cover is covered on the fixed plate, the reaction seat and the reagent placed in the reaction seat can be directly heated by the heating device arranged on the heat cover, and meanwhile, the excessive evaporation of the reagent can be avoided; meanwhile, the temperature control assembly is arranged on the bottom surface of the reagent tube seat, and can quickly control temperature rise and fall circulation, namely the temperature rise and fall circulation control of PCR can be realized;
by arranging a plurality of interface groups, each interface group corresponds to one reaction seat, two connectors of the reaction seat are respectively connected with the excitation optical fiber interface and the receiving optical fiber interface of the corresponding interface group by arranging optical fibers, and a plurality of reagent samples can be detected simultaneously; by arranging a plurality of detection components, each detection component corresponds to one detection channel, and multi-channel detection can be realized at the same time; specifically, utilize actuating mechanism drive interface dish and detect relative movement between the dish, make each interface group pass through all determine module in proper order, or make each determine module pass through all interface groups in proper order, so, each reagent sample all can detect through all determine module, and each determine module all can detect all reagent samples, realizes that all reagents can all carry out the technical purpose that the multichannel detected, can effectively improve detection efficiency.
Drawings
In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
FIG. 1 is a schematic structural diagram of an embodiment of the full-automatic fluorescence quantitative PCR analyzer of the present invention, specifically an assembly diagram of a temperature raising and lowering module and a heat dissipation assembly;
FIG. 2 is a schematic structural diagram of a temperature raising and lowering module;
FIG. 3 is a cross-sectional view of an elevating and cooling module;
FIG. 4 is a schematic view of a reagent tube holder;
FIG. 5 is a schematic view of a thermal lid assembly;
FIG. 6 is a schematic structural diagram of a fluorescence detection module;
FIG. 7 is a top view of FIG. 6;
FIG. 8 is a schematic view of the structure of the test tray;
FIG. 9 is a schematic structural diagram of an excitation light assembly;
fig. 10 is a schematic structural diagram of a light receiving module.
Description of reference numerals:
100-a base plate; 101-reagent tube seat; 102-a reaction seat; 103-a reaction chamber; 104-a fiber optic connector receptacle; 105-connection port; 106-an optical fiber; 107-step surface; 108-tube seat press plate; 109-optical fiber fixing plate; 110-a fixed plate limiting groove; 111-fiber press plate; 112-fiber cushion; 113-a platen limit slot; 114-a thermistor; 115-a temperature control element; 116-a first thermally conductive layer; 117 — second thermally conductive layer;
200-a fixed plate; 201-thermal cover; 202-a reaction zone; 203-reaction through holes; 204-a heating device; 205-a heat insulation plate; 206-heat preservation cotton; 207-locking; 208-an electromagnet;
301-interface disc; 302-detection disk; 303-excitation fiber interface; 304-a receive fiber optic interface; 305-an excitation light component; 306-a light receiving component; 307-step motor; 308-an opto-coupler sensor; 309-light barrier; 310-a conductive slip ring; 311-LED light sources; 312-a first convex lens; 313-a second convex lens; 314-a third convex lens; 315-first filter; 316-fourth convex lens; 317-a fifth convex lens; 318-PD sensor; 319-second filter;
401-middle air duct; 402-side air ducts; 403-communicating holes; 404-baffle.
Detailed Description
The present invention is further described with reference to the following drawings and specific examples so that those skilled in the art can better understand the present invention and can practice the present invention, but the examples are not intended to limit the present invention.
FIG. 1 is a schematic structural diagram of an embodiment of the full-automatic fluorescence quantitative PCR analyzer according to the present invention. The full-automatic fluorescence quantitative PCR analyzer comprises a temperature control system and a fluorescence detection module, wherein the temperature control system comprises a heating and cooling module and a hot cover assembly.
Specifically, as shown in fig. 2 and 3, the warming and cooling module includes a base plate 100 and a reagent tube holder assembly. The reagent tube seat assembly of this embodiment includes reagent tube seat 101, and the top surface of reagent tube seat 101 is equipped with reaction seat 102, is equipped with reaction chamber 103 in the reaction seat 102, and the bottom surface of reagent tube seat 101 is equipped with the temperature control assembly who is used for controlling the temperature lift circulation in the reaction chamber 103. The bottom plate 100 is provided with a pressing assembly for pressing and fixing the reagent tube holder 101. The reagent well base 101 of this embodiment has a plurality of reaction wells 102 spaced apart from each other on the top surface thereof, and all the reaction wells 102 are arranged in a one-dimensional linear array on the top surface of the reagent well base 102. The top surface of the reagent well base 101 of this embodiment is provided with 8 reaction wells 102 arranged in a one-dimensional linear array at intervals.
As shown in fig. 5, the heat cover assembly includes a fixing plate 200 and a heat cover 201 that can be covered on the fixing plate 200; a reaction zone 202 is arranged on the fixing plate 200, and a reaction through hole 203 for the reaction seat 102 to pass through is arranged in the reaction zone 202; the heating device 204 is disposed on the thermal cover 201 corresponding to the reaction area 202. The fixing plate 200 is located above the bottom plate 100, and the fixing plate 200 and the bottom plate 100 are relatively fixed; the reaction through holes 203 are arranged corresponding to the reaction sockets 102 one by one, and the reaction sockets 102 are located in the corresponding reaction through holes 203.
As shown in FIG. 6, the fluorescence detection module includes an interface disc 301, a detection disc 302, and a drive mechanism. The interface disc 301 is provided with a plurality of interface groups, and each interface group comprises an excitation optical fiber interface 303 and a receiving optical fiber interface 304. The detection plate 302 is provided with a plurality of detection elements, including an excitation light element 305 and a receiving light element 306. The driving mechanism drives the interface disc 301 and the detection disc 302 to move relatively, so that each interface group passes through all the detection assemblies in sequence, or each detection assembly passes through all the interface groups in sequence. Specifically, the following alignment relationship is satisfied between the interface group and the detection component: when the excitation light assembly 305 of the detection assembly is aligned with the excitation light interface 303 of the interface set, the receiving light assembly 306 of the detection assembly is aligned with the receiving optical fiber interface 304 of the interface set; similarly, when the receiving optical element 306 of the detection element is aligned with the receiving optical fiber interface 304 of the interface set, the excitation optical element 305 of the detection element is aligned with the excitation optical interface 303 of the interface set. In this way, in the aligned interface set and detection assembly, the light emitted from the excitation light assembly 305 is transmitted into the optical fiber 106 connected to the excitation light interface 303 corresponding thereto, and the fluorescence generated by the reagent in the reagent tube placed in the reaction chamber 103 is transmitted into the receiving light set 306 through the optical fiber 106 connected to the interface light interface 304.
An optical fiber connecting seat 104 is arranged on the outer side wall of the reaction seat 102, two connecting ports 105 are arranged on the optical fiber connecting seat 104, and the connecting ports 105 are communicated with the reaction cavity 103; the interface group is arranged corresponding to the reaction seat 102, and the two connecting ports 105 of the reaction seat 102 are respectively connected with the excitation optical fiber interface 303 and the receiving optical fiber interface 304 of the corresponding interface group through optical fibers 106.
Further, two sides of the reagent tube seat 101 are respectively provided with a step surface 107, the compressing assembly comprises tube seat pressing plates 108 respectively positioned at two sides of the reagent tube seat 101, and the tube seat pressing plates 108 are fixedly connected with the bottom plate 100 and pressed on the corresponding step surfaces 107. By providing the step surface 107, the reagent well block 101 can be firmly pressed and fixed to the bottom plate 100 by the well block pressing plate 108, and the reagent well block 101 does not need to be provided with a connecting member such as a screw connector, and the heat transfer thereof is not affected.
Further, an optical fiber fixing plate 109 is disposed on the bottom plate 100, a fixing plate limiting groove 110 for limiting the optical fiber 106 is disposed on the optical fiber fixing plate 109, and an optical fiber pressing plate 111 for pressing and fixing the optical fiber 106 in the fixing plate limiting groove 110 is disposed on the optical fiber fixing plate 109. Through set up fiber fixing plate 109 on bottom plate 100, and set up fixed plate spacing groove 110 on fiber fixing plate 109, optic fibre 106 is spacing through this fixed plate spacing groove 110, thereby can make optic fibre 106 arrange more regular, adopt optic fibre clamp plate 111 to compress tightly optic fibre 106 and fix in fixed plate spacing groove 110 simultaneously, realize compressing tightly of optic fibre 106 fixed, simultaneously can also control the bending radius of optic fibre 106 between fiber fixing plate 109 and the reaction seat 102 that corresponds, make optic fibre 106 receive the power along connector 105 axis direction, this effort can prevent to a certain extent that optic fibre 106 from producing axial displacement along connector 105. Preferably, the bottom surface of the optical fiber pressing plate 111 of the present embodiment is provided with an optical fiber cushion 112, which can not only press and fix the optical fiber 106, but also avoid damage to the optical fiber 106.
Furthermore, a tube seat pressing plate 108 located between the optical fiber fixing plate 109 and the reagent tube seat 101 is provided with a pressing plate limiting groove 113 for limiting the optical fiber 106, the pressing plate limiting groove 113 is arranged corresponding to the fixing plate limiting groove 110, and the corresponding pressing plate limiting groove 113 and the fixing plate limiting groove 110 are displaced on the same straight line, so as to further limit the bending radius of the optical fiber 106, so that the optical fiber 106 is subjected to a larger force along the axial direction of the connecting port 105.
Further, a plurality of reaction seats 102 are arranged on the top surface of the reagent tube seat 101 at intervals, and the reaction seats 102 of the present embodiment are arranged on the top surface of the reagent tube seat 101 in a one-dimensional linear array; the thermistor 114 is arranged in the reagent tube seat 101, and the thermistor 114 is positioned between two adjacent reaction seats 102. The thermistor 114 is arranged to detect the temperature, and the thermistor 114 is arranged between two adjacent reaction seats, namely, the thermistor 114 avoids the position of the reaction seat 102, so that the influence of the existence of the thermistor 114 on the heat conduction between the reagent tube seat 101 and the reaction seat 102 is reduced, the heat transfer efficiency is improved, and the temperature uniformity of each reaction seat 102 is improved.
Further, the module of heating and cooling of this embodiment still includes the excess temperature protection switch that is used for preventing reagent tube seat 101 high temperature, carries out excess temperature protection to the module of heating and cooling, avoids the high temperature.
Further, the temperature control assembly of the present embodiment includes a temperature control element 115 for controlling temperature rise and fall, and a first heat conduction layer 116 and a second heat conduction layer 117 are respectively disposed on the upper and lower sides of the temperature control element 115. Specifically, the temperature control element 115 of the present embodiment employs a semiconductor refrigerator; the first heat conducting layer 116 and the second heat conducting layer 117 are made of graphene. Temperature rise and fall are controlled by arranging the temperature control element 115, and the upper side and the lower side of the temperature control element 115 are respectively provided with the first heat conduction layer 116 and the second heat conduction layer 117, so that the rapid heat conduction between the temperature control element 115 and the reagent tube seat 101 can be realized through the first heat conduction layer 116, and when the temperature needs to be reduced, the heat can be led out through the second heat conduction layer 117, and the temperature reduction rate is improved.
Further, the periphery of the reaction area 202 is provided with heat insulation boards 205 for contacting and matching with the heating device 204, so as to insulate the heat of the temperature raising and lowering module, and facilitate the temperature control in the reaction area 202.
Further, a heating installation groove for installing the heating device 204 is formed in the thermal cover 201, the heating device is installed in the heating installation groove, a flexible elastic element is arranged between the heating device 204 and the bottom of the heating installation groove, the flexible elastic element exerts flexible elastic force on the heating device 204, the thermal cover 201 can be smoothly covered on the fixing plate 200, meanwhile, the heating device 204 can be pressed on the reaction seat 102 and the reagent tube, and heating efficiency is improved.
Further, the bottom surface of the fixing plate 200, which faces away from the hot cover 201, is provided with heat insulation cotton 206, and the heat insulation cotton 206 is provided with heat insulation through holes for the reaction seat 102 to pass through in one-to-one correspondence with the reaction through holes 203, so that the reaction seat 102 can be insulated.
Further, the connection end of the thermal cover 201 is hinged to the fixing plate 200, and a lock catch 207 is disposed between the opening/closing end of the thermal cover 201 opposite to the connection end and the fixing plate 200, so that the thermal cover 201 can be locked after the thermal cover 201 is closed. Preferably, the opening and closing end of the thermal cover 201 and/or the end of the fixing plate 200 provided with the latch 207 are provided with an electromagnet 208, and specifically, the electromagnet 208 of the present embodiment is provided on the fixing plate 200. By providing the electromagnet 208, the electromagnet 208 is electrified during operation, and a magnetic force is generated between the thermal cover 201 and the fixing plate 200 to ensure that the thermal cover 201 cannot be opened during operation.
The temperature control system of this embodiment still includes radiator unit, and radiator unit includes middle wind channel 401, and the both sides of middle wind channel 401 are equipped with respectively rather than the side wind channel 402 that is linked together, are equipped with respectively on two side wind channels 402 and go up and down temperature module and hot lid subassembly, and fixed plate 200 and bottom plate 100 are all fixed mounting in the correspondence on the side wind channel 402. In this embodiment, two side walls of the middle air duct 401 are respectively provided with communication holes 403 communicated with the two side air ducts 402, and a partition 404 located between the two communication holes 403 is provided in the middle air duct 401. Through installing on side wind channel 402 with heating and cooling module and hot cap subassembly, when temperature control assembly cooled down the reagent tube socket, the heat accessible side wind channel 402's that the bottom produced convection effect was taken away rapidly, improved cooling efficiency, through setting up baffle 404, can prevent to influence each other between two side wind channels 402.
Further, all the centers of exciting fiber optic interface 303 all are located same circular arc line I, and all the centers of receiving fiber optic interface 304 all are located a circular arc line II, and the centre of a circle coincidence of circular arc line I and circular arc line II, the radius of curvature of circular arc line I of this embodiment is greater than the radius of curvature of circular arc line II. Along the axial view direction of the excitation optical fiber interface 303 or the receiving optical fiber interface 304, the geometric centers of all the excitation optical components 305 are located on the circle on which the circular arc line i is located, and the geometric centers of all the receiving optical components 306 are located on the circle on which the circular arc line ii is located; the driving mechanism is a rotation driving mechanism for driving the interface disc 301 and the detection disc 302 to rotate relatively. The rotation driving mechanism of this embodiment includes a stepping motor 307 for driving the detection disc 302 and the interface disc 301 to rotate relatively, and the axis of the relative rotation between the detection disc 302 and the interface disc 301 passes through the centers of the circular arc line i and the circular arc line ii. The output shaft of the stepping motor 307 of the present embodiment is connected to the detection disk 302, that is, the detection disk 302 rotates synchronously with the output shaft of the stepping motor 307, and the interface disk 301 is fixedly disposed relative to the housing of the stepping motor 307. Thus, in the process of using the stepping motor 307 to drive the detection disc 302 and the interface disc 301 to rotate relatively, the excitation light assemblies 305 in all the detection assemblies sequentially pass through all the excitation optical fiber interfaces 303, the receiving light assemblies 306 sequentially pass through all the receiving optical fiber interfaces 304, each interface group corresponds to one reaction seat 102, test tubes are placed in the reaction cavities 103 of the reaction seats 102, reagents to be detected are contained in the test tubes, and thus, the same detection assembly can realize fluorescence detection of all the reagents, and the same reagent can be subjected to fluorescence detection by all the detection assemblies.
Furthermore, the included angle between the geometric centers of the excitation light assembly 305 and the receiving light assembly 306 which belong to the same detection assembly and the connecting line between the centers of the circular arc lines I is 30 degrees, and the included angle between the excitation light assembly 305 and the receiving light assembly 306 and the connecting line between the centers of the circular arc lines I is set to be 30 degrees due to the fact that the excitation light assembly 305 and the receiving light assembly 306 have certain volumes, the diameter of the detection disc 302 can be reduced, and the volume of the whole fluorescence detection module is further reduced. Specifically, the number of detection elements in this embodiment is 4, that is, the number of excitation light elements 305 and the number of reception light elements 306 are 4 respectively. The interface group of this embodiment is set to 16, and excitation fiber interface 303 and receiving fiber interface 304 are equipped with 16 respectively promptly, and correspondingly, reaction seat 102 is equipped with 16 altogether, and the module of heating and cooling is equipped with 2, is equipped with 8 reaction seats 102 on two modules of heating and cooling respectively.
Further, the fluorescence detection module of this embodiment further includes an optical coupler sensor 308 for positioning the relative position between the detection disc 302 and the interface disc 301, specifically, the optical coupler sensor 308 of this embodiment is installed on the interface disc 301, and the detection disc 302 is provided with a light blocking sheet 309 matched with the optical coupler sensor 308. The position between the interface disc 301 and the detection disc 302 is positioned by arranging the optical coupling sensor 308, so that the starting position of detection between the interface disc 301 and the detection disc 302 is determined, and the accuracy and reliability of detection are ensured. Preferably, the detection plate 302 is provided with a conductive slip ring 310, and all connection wires required by the excitation light assembly 305 and the receiving light assembly 306 are connected through the conductive slip ring 310, so that the connection wires are prevented from being wound in the rotation process of the detection plate 302.
Of course, in some embodiments, the centers of all the excitation fiber optic interfaces 303 are located on the same straight line i, the centers of all the receiving fiber optic interfaces 304 are located on a straight line ii, and the straight lines i and ii are parallel to each other; in the axial view direction of the excitation optical fiber interface 303 or the receiving optical interface 304, the geometric centers of all the excitation light assemblies 305 are located on a straight line i, and the geometric centers of all the receiving optical assemblies 306 are located on a straight line ii; the driving mechanism is a linear driving mechanism for driving the interface disc and the detection disc to move relatively. The technical purposes that the same detection assembly can realize the fluorescence detection of all reagents and the same reagent can be subjected to the fluorescence detection of all the detection assemblies can be realized, and the description is omitted.
Further, the excitation light assembly 305 includes an LED light source 311, a first convex lens 312, a second convex lens 313 and a third convex lens 314, which are sequentially disposed, a first optical filter 315 is disposed between the first convex lens 312 and the second convex lens 313, and the third convex lens 314 is located on a side of the second convex lens 313 facing the interface panel 301. The receiving optical assembly 306 of this embodiment includes a fourth convex lens 316, a fifth convex lens 317, and a PD sensor 318, which are sequentially disposed, a second optical filter 319 is disposed between the fourth convex lens 316 and the fifth convex lens 317, and the fourth convex lens 316 is located on a side of the fifth convex lens 317 facing the interface disc 301.
The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or change made by the technical personnel in the technical field on the basis of the invention is all within the protection scope of the invention. The protection scope of the invention is subject to the claims.
Claims (14)
1. The utility model provides a full-automatic fluorescence quantitative PCR analysis appearance which characterized in that: the fluorescence detection device comprises a temperature control system and a fluorescence detection module, wherein the temperature control system comprises a temperature rising and reducing module and a hot cover component;
the temperature rising and lowering module comprises a bottom plate and a reagent tube seat assembly; the reagent tube seat assembly comprises a reagent tube seat, the top surface of the reagent tube seat is provided with a reaction seat, a reaction cavity is arranged in the reaction seat, and the bottom surface of the reagent tube seat is provided with a temperature control assembly for controlling the temperature rise and fall circulation in the reaction cavity; the bottom plate is provided with a pressing assembly for pressing and fixing the reagent tube seat;
the hot cover assembly comprises a fixed plate and a hot cover capable of covering the fixed plate; a reaction zone is arranged on the fixing plate, and a reaction through hole for the reaction seat to pass through is arranged in the reaction zone; a heating device is arranged on the hot cover corresponding to the reaction zone; the fixed plate is positioned above the bottom plate, and the fixed plate and the bottom plate are relatively and fixedly arranged; the reaction through holes and the reaction seats are arranged in a one-to-one correspondence manner, and the reaction seats are positioned in the corresponding reaction through holes;
the fluorescence detection module comprises an interface disc, a detection disc and a driving mechanism, wherein a plurality of interface groups are arranged on the interface disc, and each interface group comprises an excitation optical fiber interface and a receiving optical fiber interface; the detection disc is provided with a plurality of detection components, and each detection component comprises an exciting light component and a receiving light component; the driving mechanism drives the interface disc and the detection disc to move relatively, so that each interface group sequentially passes through all the detection assemblies, or each detection assembly sequentially passes through all the interface groups;
the outer side wall of the reaction seat is provided with an optical fiber connecting seat, the optical fiber connecting seat is provided with two connecting ports, and the connecting ports are communicated with the reaction cavity; the interface group is arranged corresponding to the reaction seat, and two connectors of the reaction seat are respectively connected with the excitation optical fiber interface and the receiving optical fiber interface of the corresponding interface group through optical fibers.
2. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: step surfaces are respectively arranged on two sides of the reagent tube seat, the pressing assembly comprises tube seat pressing plates respectively positioned on two sides of the reagent tube seat, and the tube seat pressing plates are fixedly connected with the bottom plate and pressed on the corresponding step surfaces;
the bottom plate is provided with an optical fiber fixing plate, the optical fiber fixing plate is provided with a fixing plate limiting groove for limiting an optical fiber, and the optical fiber fixing plate is provided with an optical fiber pressing plate for tightly pressing and fixing the optical fiber in the fixing plate limiting groove;
and a pressing plate limiting groove for limiting the optical fiber is arranged on the tube seat pressing plate between the optical fiber fixing plate and the reagent tube seat, the pressing plate limiting groove corresponds to the fixing plate limiting groove, and the corresponding pressing plate limiting groove and the fixing plate limiting groove are displaced on the same straight line.
3. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the top surface of the reagent tube seat is provided with a plurality of reaction seats at intervals, and the reaction seats are arranged on the top surface of the reagent tube seat in a one-dimensional linear array; and a thermistor is arranged in the reagent tube seat and is positioned between two adjacent reaction seats.
4. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the temperature rising and falling module further comprises an over-temperature protection switch for preventing the reagent tube seat from being overhigh in temperature.
5. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the temperature control component comprises a temperature control element for controlling temperature to rise and fall, and a first heat conduction layer and a second heat conduction layer are respectively arranged on the upper side surface and the lower side surface of the temperature control element; the temperature control element adopts a semiconductor refrigerator; the first heat conduction layer and the second heat conduction layer are both made of graphene.
6. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the heat dissipation assembly comprises a middle air channel, two sides of the middle air channel are respectively provided with a side air channel communicated with the middle air channel, the two side air channels are respectively provided with the temperature rise and decrease module and the hot cover assembly, and the fixed plate and the bottom plate are fixedly installed on the corresponding side air channels.
7. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: heat insulation plates used for being in contact fit with the heating device are arranged on the periphery of the reaction area;
the hot cover is provided with a heating installation groove for installing a heating device, the heating device is installed in the heating installation groove, and a flexible elastic element is arranged between the heating device and the bottom of the heating installation groove.
8. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the bottom surface of the fixed plate, which faces away from the hot cover, is provided with heat-insulating cotton, and the heat-insulating cotton is provided with heat-insulating through holes which are used for the reaction seats to pass through in a one-to-one correspondence manner with the reaction through holes.
9. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the connecting end of the heat cover is hinged with the fixing plate, and a lock catch is arranged between the opening and closing end of the heat cover opposite to the connecting end and the fixing plate; and an electromagnet is arranged at the opening and closing end of the hot cover and/or the end of the fixing plate provided with the lock catch.
10. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the centers of all the excitation optical fiber interfaces are located on the same circular arc line I, the centers of all the receiving optical fiber interfaces are located on a circular arc line II, and the circle centers of the circular arc line I and the circular arc line II are overlapped; in the axial view direction of the excitation optical fiber interface or the receiving optical fiber interface, the geometric centers of all the excitation optical components are located on the circular arc line I, and the geometric centers of all the receiving optical components are located on the circular arc line II; the driving mechanism is a rotation driving mechanism for driving the interface disc and the detection disc to rotate relatively.
11. The fully automatic fluorescent quantitative PCR analyzer according to claim 10, characterized in that: the rotation driving mechanism comprises a stepping motor used for driving the detection disc and the interface disc to rotate relatively, and the axis of the relative rotation between the detection disc and the interface disc passes through the circle centers of the arc line I and the arc line II.
12. The fully automatic fluorescent quantitative PCR analyzer according to claim 10, characterized in that: the curvature radius of the circular arc line I is larger than that of the circular arc line II; and the included angle between the connecting line between the geometric centers of the exciting light assembly and the receiving light assembly which belong to the same detection assembly and the circle center of the circular arc line I is 30 degrees.
13. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the optical coupling sensor is used for positioning the relative position between the detection disc and the interface disc; and the detection plate is provided with a conductive slip ring.
14. The full-automatic fluorescent quantitative PCR analyzer according to claim 1, characterized in that: the exciting light assembly comprises an LED light source, a first convex lens, a second convex lens and a third convex lens which are sequentially arranged, a first optical filter is arranged between the first convex lens and the second convex lens, and the third convex lens is positioned on one side, facing the interface disc, of the second convex lens;
the receiving optical assembly comprises a fourth convex lens, a fifth convex lens and a PD sensor which are sequentially arranged, a second optical filter is arranged between the fourth convex lens and the fifth convex lens, and the fourth convex lens is positioned on one side of the fifth convex lens facing the interface disc.
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