CN113832020A - Optical module, thermal cycle module and PCR device suitable for PCR device - Google Patents
Optical module, thermal cycle module and PCR device suitable for PCR device Download PDFInfo
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- CN113832020A CN113832020A CN202010696808.3A CN202010696808A CN113832020A CN 113832020 A CN113832020 A CN 113832020A CN 202010696808 A CN202010696808 A CN 202010696808A CN 113832020 A CN113832020 A CN 113832020A
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- 230000003287 optical effect Effects 0.000 title claims abstract description 59
- 238000005382 thermal cycling Methods 0.000 claims abstract description 25
- 239000004065 semiconductor Substances 0.000 claims description 14
- 238000001816 cooling Methods 0.000 claims description 4
- JBRZTFJDHDCESZ-UHFFFAOYSA-N AsGa Chemical compound [As]#[Ga] JBRZTFJDHDCESZ-UHFFFAOYSA-N 0.000 claims description 3
- JMASRVWKEDWRBT-UHFFFAOYSA-N Gallium nitride Chemical compound [Ga]#N JMASRVWKEDWRBT-UHFFFAOYSA-N 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 229910044991 metal oxide Inorganic materials 0.000 claims description 3
- 150000004706 metal oxides Chemical class 0.000 claims description 3
- 238000003752 polymerase chain reaction Methods 0.000 abstract description 76
- 150000007523 nucleic acids Chemical group 0.000 description 20
- 108020004707 nucleic acids Proteins 0.000 description 13
- 102000039446 nucleic acids Human genes 0.000 description 13
- 230000003247 decreasing effect Effects 0.000 description 6
- 238000000137 annealing Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 238000011835 investigation Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000005180 public health Effects 0.000 description 4
- 238000011002 quantification Methods 0.000 description 4
- 230000003321 amplification Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000003757 reverse transcription PCR Methods 0.000 description 3
- 238000011529 RT qPCR Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 238000005549 size reduction Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000001668 nucleic acid synthesis Methods 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
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- 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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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The present invention discloses an optical module suitable for a PCR (polymerase chain reaction) apparatus, a thermal cycling module, and a PCR apparatus, the optical module comprising: an optical unit, said optical unit comprising: a light source assembly; a light sensing element; and a lens, and the surface of the lens has a grating structure; and; a sample-bearing stage, wherein the sample-bearing stage is arranged below the optical unit, and the lens is positioned between the sample-bearing stage and the light-sensitive component; wherein the light source assembly and the light sensing assembly are located on the same side or opposite sides of the stage for carrying the sample.
Description
Technical Field
The present invention relates to an optical module, a thermal cycling module, and more particularly, to an optical module suitable for a PCR (polymerase chain reaction) device, a thermal cycling module suitable for a PCR device, and a PCR device including the optical module and/or the thermal cycling module.
Background
Polymerase Chain Reaction (PCR) is a nucleic acid synthesis technique for amplifying specific nucleic acid fragments in vitro by using the principle of nucleic acid double-strand replication. By this technique, a large amount of a target gene can be amplified in a short time. PCR is a simple, cheap and reliable nucleic acid fragment amplification method, and has been widely applied in the fields of life sciences, medical diagnosis, forensic medicine, food safety, environmental detection, and the like.
A conventional PCR should consist of 20 to 35 cycles, each cycle comprising the following 3 steps:
1. denaturation: double-stranded nucleic acids are separated using high temperatures (93-98 ℃). High temperatures break the hydrogen bonds connecting the two nucleic acid strands. Prior to the first cycle, the heating is typically continued for a period of time to ensure that the template and primer are completely separated and are present only in single stranded form. The time of the previous step is 1-2 minutes, and then the machine controls the temperature to enter a circulation stage;
2. annealing or bonding, annealing: after separation of the nucleic acid duplex, the temperature is lowered so that the primer can bind to the single-stranded nucleic acid. The temperature at this stage is typically 5 ℃ below the melting point of the starter. The wrong annealing temperature may result in primers that do not bind to the template or that bind incorrectly. The time of the previous step is 1-2 minutes. The temperature of the fusion type nucleic acid polymerase in the new technology at this stage is 3-5 ℃ higher than the melting point, and only needs 5-10 seconds; and
3. extension: the nucleic acid polymerase starts to synthesize a complementary strand along the nucleic acid strand from the primer bound at the time of cooling. The temperature at this stage is dependent on the nucleic acid polymerase. The time of the preceding step depends on the polymerase and the length of the nucleic acid fragment to be synthesized.
Conventional PCR cannot provide real-time quantification of amplified nucleic acid fragments, and gel electrophoresis is necessary for further quantification. Accordingly, a Real-Time PCR, also called Quantitative Real Time Polymerase Chain Reaction (RT-PCR or qPCR), which can provide Real-Time quantification for the amplified product of nucleic acid fragments, is generated. The principle of RT-PCR is mainly that in the process of amplifying nucleic acid fragments by using PCR, a specific fluorescent agent capable of combining with double-strand nucleic acid is added, and the fluorescent agent is used for detecting the total amount of products after each PCR cycle, so that the more the double-strand nucleic acid fragments are synthesized, the higher the detected fluorescence intensity is, the more the number of cycles of PCR is, the more the nucleic acid is copied, the more the operation mechanism of simultaneously detecting fluorescence signals is, the time course of nucleic acid amplification analysis is effectively shortened, and the accuracy of nucleic acid amplification quantification is improved.
In order to detect the fluorescence emitted by the product after each PCR cycle, the light source module adopted by the existing RT-PCR device is required to be matched with a lens with radian no matter a penetrating light source module or a reflecting light source module so as to strengthen the fluorescence intensity emitted by the product after each PCR cycle is irradiated by a light source in the light source module, and the detection is carried out by a CCD photosensitive assembly.
In addition, as mentioned above, each cycle of PCR mainly includes 3 steps of high temperature denaturation, cooling annealing and low temperature extension, and each step is only different from several minutes to several seconds, the conventional thermal cycle module in the PCR device mainly uses a resistive heater or a microwave magnetron, but the heating and cooling speed of the resistive heater is too slow, consuming time and consuming power, and cannot meet the requirement of portable mobile PCR, while the microwave magnetron can quickly heat and cool, but the overall size is too large and consuming power is too large, the design will cause the size of the thermal cycle module to be too large, so that the reduction of the PCR size is difficult, the PCR device cannot be conveniently carried, and the PCR device also has a great obstacle for medical staff or public health staff to go to a laggard epidemic situation investigation in an epidemic area.
Accordingly, a light source module capable of satisfying the requirement of PCR size reduction, a thermal cycling module capable of satisfying the requirement of PCR size reduction, and a PCR device comprising the light source module and/or the thermal cycling module are needed.
Disclosure of Invention
One objective of the present invention is to disclose an optical module suitable for a PCR apparatus, comprising: an optical unit comprising: a light source assembly; a light sensing element; and a lens, the surface of the lens has a grating structure; and; a sample-bearing stage arranged below the optical unit, wherein the lens is positioned between the sample-bearing stage and the light-sensitive component; wherein the light source assembly and the light sensing assembly are located on the same side or opposite sides of the stage for carrying the sample.
As mentioned above, the light source module can be a direct-type light source module or a side-light type light source module.
In the optical module for a PCR device as described above, the light source assembly is a direct-type light source assembly, and the direct-type light source assembly includes a direct-type light emitting diode, and incident light emitted by the direct-type light emitting diode can be directly irradiated toward the stage for carrying the sample.
In the optical module for PCR device, the light source assembly is a side-light type light source assembly, and the side-light type light source assembly includes a side-light type light emitting diode and a light guide device, and the light guide device can guide the side incident light emitted from the side-light type light emitting diode to irradiate toward the stage for carrying the sample.
In the optical module for a PCR device as described above, the grating pitch size of the grating structure on the lens surface is on the micrometer scale or on the sub-micrometer scale.
In the optical module for PCR device, the light sensing element is a photodiode IC
In the above optical module for a PCR device, the photodiode IC includes a photodiode array chip and a light sensing device, the light sensing device is integrated on a surface of the photodiode array chip, and the photodiode array chip includes a photodiode electrically connected to the light sensing device.
It is another object of the present invention to disclose a PCR device comprising an optical module as described in any of the above [ h1] for use in a PCR device.
It is another object of the present invention to disclose a thermal cycling module for a PCR apparatus, the thermal cycling module comprising a semiconductor heater capable of rapidly increasing and decreasing temperature
In the thermal cycling module for a PCR device, the semiconductor heater capable of rapidly increasing and decreasing temperature is made of Laterally Diffused Metal Oxide Semiconductor (LDMOS), or gallium arsenide (GaAs) or gallium nitride (GaN) which are III-V semiconductors.
It is a further object of the present invention to disclose a PCR device comprising a thermal cycling module as described in any of the above embodiments, which is suitable for use in a PCR device.
It is a further object of the present invention to disclose a PCR apparatus, comprising: an optical module as any one of the preceding for use in a PCR device; and a thermal cycling module as described in any of the preceding embodiments adapted for use in a PCR apparatus.
Drawings
Fig. 1 shows an optical module 1000 for a PCR device according to an embodiment of the invention.
FIG. 1 'is a cross-sectional view of an optical module 1000' for use in a PCR apparatus according to another embodiment of the present invention.
FIG. 2A is a schematic cross-sectional view of a direct light source assembly 110, which can be used in the optical module 1000 for PCR device shown in FIG. 1 or the optical module 1000 'for PCR device shown in FIG. 1'.
FIG. 2B is a schematic cross-sectional view of an edge-light source assembly 110 ' suitable for use in the optical module 1000 of FIG. 1 or the optical module 1000 ' of FIG. 1 '.
FIG. 3 is a top view of a lens 120, which can be used in the optical module 1000 for PCR device shown in FIG. 1 or the optical module 1000 'for PCR device shown in FIG. 1'.
FIG. 4 is a cross-sectional view of a photo sensor assembly 130, which can be used in the optical module 1000 for the PCR device shown in FIG. 1 or the optical module 1000 'for the PCR device shown in FIG. 1'.
Fig. 5 is a block diagram of a thermal cycling module 2000 according to an embodiment of the invention.
Fig. 6 is a block diagram of a PCR apparatus 3000 including a light source module 1000 or 1000' according to an embodiment of the invention.
FIG. 7 is a block diagram of a PCR apparatus 4000 including a thermal cycling module 2000 according to an embodiment of the present invention.
Fig. 8 is a block diagram of a PCR device 5000 including a light source module 1000 or 1000' and a thermal cycling module 2000 according to an embodiment of the invention.
Wherein the symbols in the drawings are briefly described as follows:
100 optical unit
110. 110' light source assembly
111 direct type light emitting diode
113 side light type LED
115 light guide device
120 lens
125 grating structure
130 light-sensitive component
132 photodiode array chip
134 optical sensing assembly
136 photodiode
200 carrying platform
250 semiconductor heater capable of quickly raising and lowering temperature
300 sample
1000 light source module
2000 thermal cycle module
3000 ~ 5000 PCR device
Detailed Description
In order to make the disclosure of the present invention more detailed and complete, the following description is given for illustrative purposes with respect to embodiments and specific examples of the present invention; it is not intended to be the only form in which an embodiment of the invention may be practiced or utilized. The various embodiments disclosed below may be combined with or substituted for one another where advantageous, and additional embodiments may be added to one embodiment without further recitation or description.
Examples
Example one
First, please refer to fig. 1, fig. 2A to 2B, fig. 3 and fig. 4. As shown in fig. 1, the optical module 1000 for a PCR device includes: an optical unit 100 comprising: a light source assembly 110 or 110'; a light sensing element 130; and a lens 120, the surface of the lens 120 has a grating structure 125; and a sample-carrying stage 200 for carrying a sample 300, wherein the sample-carrying stage 200 is disposed below the optical unit 100, and the lens 120 is located between the sample-carrying stage 200 and the optical sensor assembly 130; the light source assembly 110 or 110' and the light sensor assembly 130 are located on opposite sides of the sample stage 200.
As shown in fig. 2A, the light source assembly 110 is a direct-type light source assembly and includes a direct-type light emitting diode 111, and incident light emitted by the direct-type light emitting diode 111 can be directly irradiated toward the sample-carrying stage 200. As shown in fig. 2B, the light source assembly 110' is a side-light type light source assembly, and includes a side-light type light emitting diode 113 and a light guide device 115, wherein the light guide device 115 can guide the side-incident light emitted by the side-light type light emitting diode 113 to irradiate towards the sample-carrying stage 200.
As shown in fig. 3, the surface of the lens 120 has a grating structure 125, and the grating pitch size of the grating structure 125 is in a micron level or a sub-micron level, so as to enhance the fluorescence intensity emitted by the product after each PCR cycle after being irradiated by the light source in the light source module.
As shown in fig. 4, the photo sensor device 130 is a photo diode IC, and the photo diode IC includes a photo diode array chip 132 and a photo sensor device 134. The photo sensor device 134 is integrated on the surface of the photodiode array chip 132, and the photodiode array chip 132 includes a photodiode 136 therein, and the photodiode 136 is electrically connected to the photo sensor device 134.
In the optical module 1000 suitable for the PCR device, since the lens 120 has no curvature, and the used light sensing element 130 is a photodiode IC, and the photodiode IC includes a photodiode array chip 132 and a light sensing element 134 integrated on the surface of the photodiode array chip 132, a smaller light-gathering distance of the optical module 1000 can be provided, so as to achieve the purpose of downsizing the optical module.
Example two
First, please refer to fig. 1', fig. 2A-2B, fig. 3 and fig. 4. As shown in fig. 1 ', the optical module 1000' for a PCR device includes: an optical unit 100' comprising: a light source assembly 110 or 110'; a light sensing element 130; and a lens 120, the surface of the lens 120 has a grating structure 125; and a sample-carrying stage 200 for carrying a sample 300, wherein the sample-carrying stage 200 is disposed below the optical unit 100, and the lens 120 is located between the sample-carrying stage 200 and the optical sensor assembly 130; the light source assembly 110 or 110' and the light sensor assembly 130 are located on the same side of the sample stage 200.
As shown in fig. 2A, the light source assembly 110 is a direct-type light source assembly and includes a direct-type light emitting diode 111, and incident light emitted by the direct-type light emitting diode 111 can be directly irradiated toward the sample-carrying stage 200. As shown in fig. 2B, the light source assembly 110' is a side-light type light source assembly, and includes a side-light type light emitting diode 113 and a light guide device 115, wherein the light guide device 115 can guide the side-incident light emitted by the side-light type light emitting diode 113 to irradiate towards the sample-carrying stage 200.
As shown in fig. 3, the surface of the lens 120 has a grating structure 125, and the grating pitch size of the grating structure 125 is in a micron level or a sub-micron level, so as to enhance the fluorescence intensity emitted by the product after each PCR cycle after being irradiated by the light source in the light source module.
As shown in fig. 4, the photo sensor device 130 is a photo diode IC, and the photo diode IC includes a photo diode array chip 132 and a photo sensor device 134. The photo sensor device 134 is integrated on the surface of the photodiode array chip 132, and the photodiode array chip 132 includes a photodiode 136 therein, and the photodiode 136 is electrically connected to the photo sensor device 134.
In the optical module 1000 'for the PCR apparatus, since the lens 120 has no curvature, and the used light sensing element 130 is a photodiode IC, and the photodiode IC includes a photodiode array chip 132 and a light sensing element 134 integrated on the surface of the photodiode array chip 132, a smaller light-gathering distance of the optical module 1000' can be provided, so as to achieve the purpose of downsizing the optical module.
EXAMPLE III
Referring to fig. 5, a third embodiment of the present invention discloses a thermal cycling module 2000 suitable for a PCR apparatus, wherein the thermal cycling module includes a semiconductor heater 250 capable of rapidly increasing and decreasing temperature. The semiconductor heater 250 capable of rapidly increasing and decreasing temperature is made of Lateral Diffused Metal Oxide Semiconductor (LDMOS) or III-V semiconductor gallium arsenide (GaAs) or gallium nitride (GaN), for example, but not limited thereto.
In the third embodiment, the thermal cycling module 2000 adapted to the PCR apparatus can achieve the purpose of reducing the size of the thermal cycling module because the semiconductor heater 250 capable of rapidly increasing and decreasing the temperature is used, and the overall size is reduced compared to the conventional microwave magnetron heater in addition to rapidly increasing and decreasing the temperature in each cycle step of the PCR.
Example four
As shown in fig. 6, a PCR device 3000 including the light source module 1000 according to the first embodiment or the light source module 1000' according to the second embodiment is disclosed in the fourth embodiment. In the PCR device 3000 of the fourth embodiment, the light source module 1000 or 1000' with reduced size as shown in the first or second embodiment is adopted, so that the size of the PCR device can be greatly reduced, and the PCR device can be conveniently carried, which is helpful for medical staff or public health staff to go to a laggard epidemic area for epidemic situation investigation.
EXAMPLE five
As shown in FIG. 7, the fifth embodiment discloses a PCR apparatus 4000 including a thermal cycling module 2000 as shown in the third embodiment. In the PCR device 4000 disclosed in the fifth embodiment, since the thermal cycling module 2000 with a reduced size is adopted as shown in the third embodiment, the size of the PCR device can be greatly reduced, so as to achieve the purpose of being conveniently carried, thereby being greatly helpful for medical staff or public health staff to go to a laggard epidemic area for epidemic investigation.
EXAMPLE six
As shown in fig. 8, the sixth embodiment discloses a PCR device 5000 including the light source module 1000 shown in the first embodiment or the light source module 1000' shown in the second embodiment and the thermal cycling module 2000 shown in the third embodiment. In the PCR device 5000 disclosed in the sixth embodiment, the light source module 1000 or 1000' with reduced size shown in the first embodiment or the second embodiment and the thermal cycling module 2000 with reduced size shown in the third embodiment are adopted, so that the size of the PCR device can be greatly reduced, the PCR device can be conveniently carried, and the PCR device is greatly helpful for medical staff or public health staff to go to a laggard epidemic situation investigation in an epidemic area.
The above description is only for the preferred embodiment of the present invention and should not be construed as limiting the scope of the present invention, and any person skilled in the art can make further modifications and variations without departing from the spirit and scope of the present invention.
Claims (12)
1. An optical module adapted for use in a PCR device, comprising:
an optical unit, said optical unit comprising:
a light source assembly;
a light sensing element; and
a lens, wherein the surface of the lens is provided with a grating structure; and
a sample-bearing stage, wherein the sample-bearing stage is arranged below the optical unit, and the lens is positioned between the sample-bearing stage and the light-sensitive component;
wherein the light source assembly and the light sensing assembly are located on the same side or opposite sides of the stage for carrying the sample.
2. The optical module for a PCR apparatus as set forth in claim 1, wherein the light source assembly is a direct-type light source assembly or a side-light type light source assembly.
3. The optical module of claim 2, wherein the light source assembly is a direct-type light source assembly, and the direct-type light source assembly comprises a direct-type light emitting diode, and incident light emitted by the direct-type light emitting diode is directly irradiated toward the stage for carrying the sample.
4. The optical module of claim 2, wherein the light source assembly is a side-light type light source assembly, and the side-light type light source assembly comprises a side-light type light emitting diode and a light guide device, and the light guide device guides the side-incident light emitted by the side-light type light emitting diode to irradiate towards the sample-carrying stage.
5. The optical module for a PCR device according to claim 1, wherein the grating pitch size of the grating structure on the lens surface is on the micrometer scale or on the sub-micrometer scale.
6. The optical module of claim 1, wherein the light sensor is a photodiode IC.
7. The optical module of claim 6, wherein the photodiode IC comprises a photodiode array chip and a light sensing device, the light sensing device is integrated on a surface of the photodiode array chip, the photodiode array chip comprises a photodiode, and the photodiode is electrically connected to the light sensing device.
8. A PCR device comprising an optical module for a PCR device according to any one of claims 1 to 7.
9. A thermal cycling module suitable for a PCR device is characterized in that the thermal cycling module comprises a semiconductor heater capable of rapidly heating and cooling.
10. The thermal cycling module for use in a PCR device according to claim 9, wherein the semiconductor heater capable of rapid temperature rise and fall is made of Laterally Diffused Metal Oxide Semiconductor (LDMOS) or gallium arsenide (GaAs) or gallium nitride (GaN) which are group III-V semiconductors.
11. A PCR device comprising an optical module suitable for use in a PCR device according to claim 9 or 10.
12. A PCR device, comprising:
an optical module for a PCR device according to any one of claims 1 to 7; and
a thermocycling module according to claim 9 or claim 10 adapted for use in a PCR device.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| TW109208136 | 2020-06-24 | ||
| TW109208136U TWM602562U (en) | 2020-06-24 | 2020-06-24 | An optical module for a pcr apparatus and a pcr apparatus comprising the same |
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| CN113832020A true CN113832020A (en) | 2021-12-24 |
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| TWM602562U (en) * | 2020-06-24 | 2020-10-11 | 傅宗民 | An optical module for a pcr apparatus and a pcr apparatus comprising the same |
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