CN112827524A - Thermal cycling device - Google Patents
Thermal cycling device Download PDFInfo
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- CN112827524A CN112827524A CN202010792851.XA CN202010792851A CN112827524A CN 112827524 A CN112827524 A CN 112827524A CN 202010792851 A CN202010792851 A CN 202010792851A CN 112827524 A CN112827524 A CN 112827524A
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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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Abstract
An embodiment of the present invention provides a thermal cycle apparatus, including: a sample block (1) provided with a blind hole array; and N heating and cooling devices (2), N being a positive integer greater than 2; wherein the upper parts of all blind holes (11) are connected, N heating and cooling devices (2) respectively heat and/or cool the bottoms of the blind holes (11) which are positioned in N different areas of the sample block (1), and each area comprises one or more blind holes (11). According to the embodiment of the invention, the upper parts of the blind holes in the sample block are connected, and the bottoms of the blind holes in different areas of the sample block are heated and/or cooled by the N heating and cooling devices, so that the heat transfer effect between the blind holes in different areas of the sample block is reduced, and the accuracy of temperature control on different areas of the sample block and the repeatability of an experiment are improved.
Description
Technical Field
The invention relates to the technical field of reaction vessels for performing chemical reactions or biological reactions, in particular to a thermal cycling device.
Background
In exploring optimal reaction temperatures and optimal reaction times for biological or chemical samples, it is often necessary to set and perform a series of temperature cycles on the biological or chemical sample from which an optimal temperature and time combination is selected.
The prior art device for performing the series of temperature cycles is a Polymerase Chain Reaction (PCR) nucleic acid amplification apparatus, which may be referred to as a PCR apparatus or a thermal cycling apparatus. The thermal cycling device comprises a sample block on which a blind hole array is uniformly arranged, and a heating device and a cooling device are arranged below two opposite sides of the sample block. And placing the PCR plate or the PCR tube filled with the biological sample into the blind hole of the sample block, and enabling the biological sample to undergo a plurality of temperature cycles in the thermal cycling device by presetting heating temperature, heating time, cooling temperature and cooling time. When heating the sample piece, two heating device independently heat up to different preset temperature, heat rather than the sample piece both sides that correspond, and the part between the sample piece both sides can realize the ladder temperature between two preset temperatures through heat-conduction, and the heat conducts to biological sample through the blind hole and the PCR board of sample piece. When the sample block is cooled, the cooling device moves to the lower parts of two opposite sides of the sample block to execute a heat dissipation process. The temperature profile on the sample block is similar to the heating process and will not be described further herein.
Therefore, the temperature of the two sides of the sample block corresponding to the heating device and the cooling device can be accurately controlled to the preset temperature, and the temperature of the part between the two sides of the sample block influences the repeatability of the experiment due to the uncontrollable heat conduction effect.
Disclosure of Invention
The embodiment of the invention provides a thermal cycling device, and aims to solve the problems of accuracy and experiment repeatability of temperature control of each part of a sample block in the prior art.
In a first aspect, a thermal cycler is provided, comprising:
a sample block (1) provided with a blind hole array; and
n heating and cooling devices (2), N being a positive integer greater than 2;
wherein the upper parts of all blind holes (11) are connected, N heating and cooling devices (2) respectively heat and/or cool the bottoms of the blind holes (11) which are positioned in N different areas of the sample block (1), and each area comprises one or more blind holes (11).
According to the embodiment of the invention, the upper parts of the blind holes in the sample block are connected, and the bottoms of the blind holes in different areas of the sample block are heated and/or cooled by the N heating and cooling devices, so that the heat transfer effect between the blind holes in different areas of the sample block is reduced, and the accuracy of temperature control on different areas of the sample block and the repeatability of an experiment are improved.
Drawings
The foregoing and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is an exploded view of a thermal cycler according to one embodiment of the present invention;
FIG. 2 is a top view and a bottom view of a sample block provided by an embodiment of the present invention with the blind holes disconnected at the bottom;
FIG. 3 is a top and bottom view of a sample block with the bottoms of blind holes connected in a column according to an embodiment of the present invention;
FIG. 4 is a top and bottom view of sample blocks with the bottoms of blind holes connected in two rows according to an embodiment of the present invention;
FIG. 5 is a top and bottom view of sample blocks with the bottoms of blind holes connected in two rows according to another embodiment of the present invention;
FIG. 6 is an exploded view of a thermal cycler according to a second embodiment of the present invention;
FIG. 7 is a top view of a sample block provided in accordance with a second embodiment of the present invention;
fig. 8 is an exploded view of a thermal cycler according to a third embodiment of the present invention;
fig. 9 is a schematic view of a thermal cycler according to a third embodiment of the present invention.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar modules or modules having the same or similar functionality throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention. On the contrary, the embodiments of the invention include all changes, modifications and equivalents coming within the spirit and terms of the claims appended hereto.
According to the embodiment of the invention, the upper parts of the blind holes in the sample block are connected, and the bottoms of the blind holes in different areas of the sample block are heated and/or cooled by the N heating and cooling devices, so that the heat transfer effect between the blind holes in different areas of the sample block is reduced, and the accuracy of temperature control on different areas of the sample block and the repeatability of an experiment are improved.
Example one
Fig. 1 is an exploded view of a thermal cycler according to an embodiment of the present invention. As shown in fig. 1, the thermal cycler includes a sample block 1, N heating and cooling devices 2.
In the embodiment of the invention, the sample block 1 is provided with blind holes 11 which are uniformly arranged at equal intervals to form a blind hole array. The blind holes 11 refer to the hole grooves on the sample block 1 for placing the PCR tubes, and the size and the distance of the blind holes are set to be matched with standard or non-standard PCR plates and PCR tubes, so that the PCR plates filled with biological samples can be integrally placed in the blind hole array, and the inner walls of the blind holes 11 are close to the outer walls of the tubes of the PCR plates as much as possible, thereby being beneficial to heat conduction. The biological sample is a substance to undergo a thermal cycling reaction, and includes, but is not limited to, oligonucleotides as primers, dNTP mixture, Taq DNA polymerase, PCR buffer, and the like. The upper parts of all blind holes 11 are connected. The connecting part is as close as possible to the opening of the blind hole or is parallel and level to the opening of the blind hole and far away from the bottom of the blind hole. The connecting portion and each blind hole 11 can be integrally formed by using the same material to form a whole sample block 1.
As one embodiment of the present invention, the sample block 1 includes a first substrate 12 protruding toward the outer periphery at the upper portion of the outermost peripheral blind via 11 of the blind via array. The first substrate 12 may be formed by integrally molding the connecting portion and the blind holes 11, with the connecting portion extending toward the outer periphery. The thermal cycler also includes a press fitting 3. The press member 3 presses the sample block 1 against the N heating and cooling devices 2 via the first substrate 12, with the array of blind holes emerging from a window in the middle of the press member 3.
The material of the sample block 1 as a whole may be selected from materials having good thermal conductivity. Preferably, it is an aluminum alloy, aluminum, copper, silver, or the like.
N heating and cooling devices 2 are located below the sample block 1, N being a positive integer greater than 2. As shown in fig. 1, N is six, but is not limited thereto. The heating and cooling device 2 includes, but is not limited to, TEC (semiconductor Cooler), heater wire, heater rod, air conduction device or liquid conduction device, etc. In an embodiment of the invention, six TECs each independently heat and/or cool the bottom of a blind hole 11 located in six different regions of the sample block 1, each region comprising one or more blind holes 11, to a target temperature. As shown in fig. 1, one TEC heats and/or cools the bottom of two columns (12) of blind holes 11.
Fig. 2 is a top view and a bottom view of the sample block 1 when the bottoms of all the blind holes 11 are not connected, in this case, the sample block 1 is divided into different regions in the shape of TEC. The blind hole 11 corresponding to each TEC is a heating and/or cooling area thereof.
Considering the problem of temperature uniformity of the blind holes 11 in different areas, as an embodiment of the present invention, the bottom of each blind hole may be connected in a manner that the bottoms of M columns of blind holes shown in fig. 3, 4, and 5 are connected, where M is a positive integer greater than or equal to 1 and less than or equal to N. FIG. 3 is a top view and a bottom view of the sample block 1 with the bottoms of the blind holes 11 connected in a row; FIG. 4 is a top view and a bottom view of the sample block 1 with the bottoms of the blind holes 11 connected in two rows; fig. 5 is a top view and a bottom view of the sample block 1 when the bottoms of the blind holes 11 are connected in two rows, and a groove is provided at the connecting portion, the groove not penetrating the connecting portion. The connecting part at the bottom of the blind hole 11 and the sample block 1 can be integrally formed by using the same material to form a whole sample block 1. Based on different connection modes of the bottoms of the blind holes 11, the sample block 1 is divided into different areas, and correspondingly, the plurality of TECs respectively and independently heat and/or cool the bottoms of the blind holes in the different areas. By the mode, different heating temperatures, different heating times and different heating rates can be set for different areas of the sample block 1, the volume of the biological sample in the PCR tube can be changed according to the division of the different areas of the sample block 1, the primers in the PCR tube can be changed, and the optimal conditions for carrying out experiments can be explored.
Since the material of the sample block 1 is a metal having good thermal conductivity, the outer walls of all the blind holes 11 are not connected except for the connecting portion at the upper part of the blind hole 11 and the connecting portion at the bottom of the blind hole 11 in order to reduce the overall weight of the sample block 1.
In the experimental process, the sample block 1 contains a PCR plate or a PCR tube filled with biological samples, the biological samples are mainly distributed at the positions corresponding to the lower parts of the blind holes 11, the upper parts of the blind holes in the sample block are connected, and the bottoms of the blind holes in different areas of the sample block are respectively heated and/or cooled by N heating and cooling devices, so that the heat transfer effect among the blind holes in different areas of the sample block is reduced, the temperature of the blind holes in each area of the sample block is uniform, the accuracy of temperature control on different areas of the sample block and the repeatability of the experiment are improved, and the method is suitable for optimizing the multiple PCR nucleic acid amplification process.
Example two
In the embodiment of the present invention, the basic structure of the thermal cycling apparatus is the same as that of the first embodiment, and the same components as those of the first embodiment are denoted by the same reference numerals as those of the first embodiment, including all the features described in the first embodiment, and will not be described again here.
Fig. 6 is an exploded view of a thermal cycler according to a second embodiment of the present invention. FIG. 7 is a top view of a sample block provided in accordance with a second embodiment of the present invention. With reference to fig. 6 and 7, the thermal cycler includes a sample block 1, N heating and cooling devices 2, and a press member 3. The sample block 1 further comprises an array of through holes arranged in a plane contiguous to the upper part of the blind holes 11. Through holes 13 are provided between the blind holes 11 at the connecting portion of the upper portions of the blind holes 11. In the embodiment of the present invention, as shown in fig. 7, one through hole 13 is provided in the middle area surrounded by every four blind holes 11. Further, a plurality of through holes 13 are provided in this region and/or between every two blind holes 11. Thereby, the metal heat transfer in this region is changed to air heat transfer, reducing the heat conduction effect between the blind holes 11. In order to further reduce the heat transfer effect, the thermal cycling device further comprises a heat insulating material filled in the through hole 13 and a heat insulating member 4 filled in the outer wall of the blind hole 11. The thermal conductivity of these insulating materials and insulation 4 should be lower than that of air.
In the embodiment of the present invention, the sample block 1 includes the second substrate 14 protruding toward the outer periphery at the bottom of the outermost peripheral blind hole of the blind hole array. The first substrate 14 and the blind holes 11 are integrally molded by using the same material to form the press-fixing member 3, the sample block 1 is pressed on the N heating and cooling devices 2 through the second substrate 14, and the blind hole array is exposed from the window in the middle of the press-fixing member 3.
According to the embodiment of the invention, the upper parts of the blind holes in the sample block are connected, the connecting part of the blind holes is punched, the heat insulating material is filled in the gap of the sample block, and the bottoms of the blind holes in different areas of the sample block are respectively heated and/or cooled through the N heating and cooling devices, so that the heat transfer effect between the blind holes in different areas of the sample block is reduced, and the accuracy of temperature control on different areas of the sample block and the repeatability of an experiment are improved.
EXAMPLE III
In the embodiment of the present invention, the basic structure of the thermal cycling apparatus is the same as that of the first embodiment or the second embodiment, and the same components as those of the first embodiment or the second embodiment are denoted by the same reference numerals, including all the features described therein, and are not described again here.
Fig. 8 is an exploded view of a thermal cycler according to a third embodiment of the present invention. Fig. 9 is a schematic view of a thermal cycler according to a third embodiment of the present invention. Referring to fig. 8 and 9, the thermal cycling apparatus includes a sample block 1, N heating and cooling apparatuses 2, a press fitting member 3, a first heat conductive element 5, a TEC spacer 6, a second heat conductive element 7, a heat sink 8, a thermal cover 9, and a bottom case 10.
The bottom case 10 serves to carry all the weight of the thermal cycler. The heat sink 8 is fixed in the bottom case 10, and the upper portion thereof is a flat plane. A single piece or N second heat conducting elements 7 lie flat on the upper plane of the heat sink 8, with the lower surface of the second heat conducting element 7 in contact with the upper surface of the heat sink 8. The N TECs with the upper surfaces and the lower surfaces coated with the heat conduction silicone grease are correspondingly placed on the N second heat conduction elements 7 respectively, and the lower surfaces of the TECs are in contact with the upper surfaces of the second heat conduction elements 7. The N TECs are separated by the hollowed-out TEC isolating piece 6, the TEC isolating piece 6 is a high-temperature-resistant heat-insulating material, and the hollowed-out parts are determined according to the number and the size of the TECs, so that the TECs are mutually thermally isolated. The N first heat conducting elements 5 are correspondingly arranged on the N TECs respectively, and the lower surfaces of the first heat conducting elements 5 are in contact with the upper surfaces of the TECs. The first and second heat conducting elements 5 and 7 include, but are not limited to, heat conducting materials such as heat gasket, aluminum foil, copper foil, or silver foil, which have high heat conduction efficiency. The sample block 1 is placed on the N first heat-conducting elements 5 with the bottom of each blind hole 11 in contact with the upper surface of the first heat-conducting element 5. The pressing piece 3 is pressed on the first substrate 12 of the sample block 1, and then the pressing piece 3 is connected to the heat sink 8 through screws, so as to further press the sample block 1, the first heat conducting element 5, the TEC, and the second heat conducting element 7 on the heat sink 8. The gaps between the bottom of each blind hole 11 and the upper surface of the TEC are filled with the heat conducting silicone grease and the first heat conducting element 5 under pressure, and the gaps between the lower surface of the TEC and the upper surface of the heat dissipating device 8 are filled with the heat conducting silicone grease and the second heat conducting element 7 under pressure. The seamless design can reduce thermal resistance and increase heat conduction efficiency.
The thermal cycler can be used in a variety of embodiments for controlling the temperature of the bottom of blind holes in different areas of the block 1. As an embodiment of the invention, the PCR tubes contained in all the blind holes 11 of the sample block 1 react at the same temperature, and the N TECs heat or refrigerate according to the same temperature instruction so as to reach the same temperature of all the blind holes 11. As another embodiment of the invention, PCR tubes contained in the blind holes 11 in different areas of the sample block 1 react according to different temperatures, so that the temperature of the blind holes 11 in different areas of the sample block 1 changes in a gradient manner in the same thermal cycle process. As a further embodiment of the invention, some of the blind holes 11 are selected from different areas of the sample block 1 to accommodate PCR tubes and to perform reactions at different temperatures. The larger the distance between the selected blind holes 11 is, the better the distance between the selected blind holes 11 is, the adjacent blind holes 11 can be subjected to temperature compensation (heating or cooling) by adopting the TEC, so that the thermal influence between the selected adjacent blind holes 11 is reduced.
As shown in fig. 9, the thermal cycling device further includes a heating and cooling control system, a central control system, and a heat dissipation control system. In the embodiment of the present invention, different high-sensitivity thermal sensors (not shown in the figure) are respectively connected to different regions of the sample block 1.
When heating is carried out, the central control system sends out a sample block heating command, and the command is transmitted to the heating and cooling control system through the central and heating and cooling control loops. After being processed, the heating and cooling control system acts on the N TECs through the TEC control loop, the upper surfaces of the TECs start to heat, and the lower surfaces of the TECs correspondingly start to refrigerate. The temperature sensor transmits the detected temperature back to the heating and cooling control system through the TEC control loop, the heating and cooling control system judges whether the target temperature is reached, and then the TEC is controlled to continue heating or stop heating through the TEC control loop. To continue heating, the TEC continuously conducts heat to the sample block 1, thereby continuously providing heat to the PCR plate or the biological sample within the PCR tube. This process requires several cycles.
When cooling is carried out, the central control system sends out a sample block cooling instruction, and the instruction is transmitted to the heating and cooling control system through the central and heating and cooling control loop and is transmitted to the heat dissipation control system through the central and heat dissipation control loop. The heating and cooling control system is processed and then acts on the N TECs through the TEC control loop, the upper surfaces of the TECs start to refrigerate, and the lower surfaces of the TECs correspondingly start to transfer heat to the heat dissipation device 8. The temperature sensor transmits the detected temperature back to the heating and cooling control system through the TEC control loop, the heating and cooling control system judges whether the target temperature is reached, and then the TEC is controlled to continue cooling or stop cooling through the TEC control loop. To continue cooling, the TEC continuously extracts heat from the sample block 1, thereby continuously cooling the biological sample in the PCR plate or PCR tube. The heat dissipation control system is processed and then acts on the heat dissipation device 8 through the heat dissipation control loop, and heat is transferred to the outside of the heat circulation device. This process requires several cycles.
The inside of the thermal cover 9 is provided with a heating device (not shown in the figure) and a high-sensitivity heat-sensitive sensor (not shown in the figure). When heating is carried out, the central control system sends out a heating command of the heating cover, and the command is transmitted to the heating and cooling control system by the central control system and the heating and cooling control loop. The heating and cooling control system is processed and then acts on the hot lid 9 through the hot lid heating control loop, and the heating device in the hot lid 9 starts to generate heat. The temperature sensor sends the detected temperature back to the heating and cooling control system through the hot cover heating control loop, the heating and cooling control system judges whether the target temperature is reached, and then the heating device in the hot cover 9 is controlled to continue heating or stop heating through the hot cover heating control loop. If heating is to be continued, the heating means in the thermal cap 9 continuously conducts heat to the PCR plate or the nozzle of the PCR tube. This process requires several cycles.
The heating and cooling control system, the central control system and the heat dissipation control system may be integrated into one control system, which is not limited herein.
According to the embodiment of the invention, the upper parts of the blind holes in the sample block are connected, and the bottoms of the blind holes in different areas of the sample block are heated and/or cooled by the N heating and cooling devices, so that the heat transfer effect between the blind holes in different areas of the sample block is reduced, and the accuracy of temperature control on different areas of the sample block and the repeatability of an experiment are improved.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (10)
1. A thermal cycler, comprising:
a sample block (1) provided with a blind hole array; and
n heating and cooling devices (2), N being a positive integer greater than 2;
wherein the upper parts of all blind holes (11) are connected, the N heating and cooling devices (2) respectively heat and/or cool the bottom parts of the blind holes (11) which are positioned in N different areas of the sample block (1), and each area comprises one or more blind holes (11).
2. A thermocycling device according to claim 1, wherein the bottoms of all blind holes (11) are not connected.
3. The thermal cycler apparatus of claim 1, wherein the bottom of the M series of blind holes (11) are connected, M being a positive integer greater than or equal to 1 and less than or equal to N.
4. A thermocycling device according to any one of claims 1 to 3, wherein the sample block (1) further comprises an array of through holes arranged in a contiguous face at the upper part of the blind holes (11).
5. The thermal cycler apparatus of claim 4, further comprising an insulating material filled in the through-holes (13).
6. The thermal cycler apparatus of claim 4, further comprising a thermal insulator (4) filled in an outer wall of the blind hole (11).
7. The thermal cycler apparatus of claim 1, further comprising a hold down (3), the hold down (3) pressing the sample block (1) against the N heating and cooling devices (2).
8. The thermal cycler apparatus of claim 7, wherein the sample block (1) further comprises a first base plate (12) protruding peripherally above the outermost blind holes (11) of the array; the press (3) presses the sample block (1) against the N heating and cooling devices (2) via the first base plate (12).
9. The thermal cycler apparatus of claim 7, wherein the sample block (1) further comprises a second substrate (14) protruding peripherally at the bottom of the outermost blind hole (11) of the array; the pressing part (3) presses the sample block (1) on the N heating and cooling devices (2) through the second substrate (14).
10. The thermal cycler apparatus of claim 1, wherein the heating and cooling apparatus (2) is a semiconductor Cooler (TEC).
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CN202010792851.XA CN112827524A (en) | 2020-08-10 | 2020-08-10 | Thermal cycling device |
PCT/CN2021/103962 WO2022033226A1 (en) | 2020-08-10 | 2021-07-01 | Thermal cycling device |
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CN202010792851.XA CN112827524A (en) | 2020-08-10 | 2020-08-10 | Thermal cycling device |
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