CN113358650B - 96 hole micropore board reader - Google Patents
96 hole micropore board reader Download PDFInfo
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- CN113358650B CN113358650B CN202110610917.3A CN202110610917A CN113358650B CN 113358650 B CN113358650 B CN 113358650B CN 202110610917 A CN202110610917 A CN 202110610917A CN 113358650 B CN113358650 B CN 113358650B
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- light source
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- pore plate
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- 239000013307 optical fiber Substances 0.000 claims abstract description 52
- 239000011148 porous material Substances 0.000 claims abstract description 33
- 238000004321 preservation Methods 0.000 claims abstract description 28
- 230000003287 optical effect Effects 0.000 claims abstract description 21
- 238000012545 processing Methods 0.000 claims abstract description 17
- 230000005540 biological transmission Effects 0.000 claims abstract description 12
- 230000003993 interaction Effects 0.000 claims abstract description 11
- 239000007788 liquid Substances 0.000 claims description 26
- 238000001514 detection method Methods 0.000 claims description 20
- 238000010438 heat treatment Methods 0.000 claims description 9
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims description 4
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 claims description 4
- 229910052782 aluminium Inorganic materials 0.000 claims description 4
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 4
- 229910052802 copper Inorganic materials 0.000 claims description 4
- 239000010949 copper Substances 0.000 claims description 4
- 229910052751 metal Inorganic materials 0.000 claims description 4
- 239000002184 metal Substances 0.000 claims description 4
- 150000002739 metals Chemical class 0.000 claims description 4
- 229910052709 silver Inorganic materials 0.000 claims description 4
- 239000004332 silver Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 abstract description 5
- 238000012423 maintenance Methods 0.000 abstract description 4
- 238000007405 data analysis Methods 0.000 abstract 1
- 238000002965 ELISA Methods 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 6
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 6
- 238000012742 biochemical analysis Methods 0.000 description 5
- 239000000758 substrate Substances 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 229940109239 creatinine Drugs 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 210000002700 urine Anatomy 0.000 description 3
- 230000007547 defect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 238000002558 medical inspection Methods 0.000 description 1
- 238000004452 microanalysis Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- OXNIZHLAWKMVMX-UHFFFAOYSA-N picric acid Chemical compound OC1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O OXNIZHLAWKMVMX-UHFFFAOYSA-N 0.000 description 1
- 238000012123 point-of-care testing Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- WFKWXMTUELFFGS-UHFFFAOYSA-N tungsten Chemical compound [W] WFKWXMTUELFFGS-UHFFFAOYSA-N 0.000 description 1
- 229910052721 tungsten Inorganic materials 0.000 description 1
- 239000010937 tungsten Substances 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Classifications
-
- 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
- G01N21/84—Systems specially adapted for particular applications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
Abstract
The invention provides a 96-hole micro-pore plate reader, which comprises an interaction and central processing module, a light source module, a temperature control system, an image transmission system and an image acquisition system, wherein light rays emitted by the light source module according to set parameters pass through micro-pore plates arranged on a micro-pore plate heat preservation frame, are focused on an optical fiber bundle by a condensing lens in 96 through holes, are converged at an optical fiber bundling device by an optical fiber bundle adjusting light path, are finally converted into image signals by a camera through an optical filter selected on an optical filter selector and the image acquisition lens to be transmitted to the interaction and central processing module for data analysis or sharing, and the temperature control system is designed to emit heat generated by the light source module and maintain a constant temperature environment for the micro-pore plates. The reader provided by the invention can rapidly, accurately and conveniently acquire the concentration of the object to be detected in the solution; the reaction process of each hole of the micro-hole plate can be synchronously observed in real time; the cost is low, the operation is simple, the purchase and maintenance cost of the instrument can be saved, and the popularization and the use in basic units are facilitated.
Description
Technical Field
The invention belongs to the field of medical inspection or chemical analysis, and relates to a microplate reader, in particular to a 96-hole microplate reader.
Background
With the development of biochemical analysis technologies such as ELISA (enzyme-linked immunosorbent assay) technology, the analysis technology has the characteristics and trends of high throughput, high sensitivity, low dosage consumption, multiple marks and the like, so that a microplate becomes a standard device in the field of high-throughput biochemical analysis, and a detection device represented by an enzyme-labeled instrument also becomes a basic instrument of biochemical analysis. Further expansion and diversification of biochemical analysis technology also put more complex requirements on corresponding detection instruments, and application of the traditional enzyme-labeled instrument in numerous scenes has been limited. In recent years, in order to meet the demands of continuous development of high-throughput biological analysis technology, a multifunctional microplate detector has emerged. On the premise of sacrificing part of performance indexes, the detector uses a higher-density micro-pore plate as a carrier, so that the unit area can be compatible with more sample numbers, and can perform more micro analysis while saving a large amount of reagent consumption; the automatic analysis processing capacity is enhanced, and the full-automatic process integrating sample adding, detection, result transmission and data processing until the micro-pore plate is washed can be realized; based on the original single function, the multifunctional integrated detection can be synchronously realized by matching with a new analysis detection technology. However, the light sources of the existing enzyme-labeled instrument and microplate reader are usually tungsten lamps, so that the defects of high requirements on the use environment, low detection speed, high price, high maintenance cost and high difficulty of the existing enzyme-labeled instrument and microplate reader are caused by large whole volume and large mass of the instrument, and the application scene of detection means such as ELISA (enzyme-Linked immuno sorbent assay) detection technology and the like on a base layer is greatly limited.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides a 96-hole microplate reader, and solves the problems that: the enzyme label instrument and the microplate reader are not portable enough; the detection speed of the enzyme label instrument and the microplate reader is low; the enzyme label instrument and the microplate reader have complex structures and high manufacturing and maintenance costs.
The aim of the invention is achieved by the following technical scheme: the utility model provides a 96 hole micropore board reader, this reader includes mutual and central processing module, light source module, accuse temperature system, image transmission system and image acquisition system, mutual and central processing module pass through the wire simultaneously with light source module, accuse temperature system and image acquisition system link to each other, the light source radiator of accuse temperature system is hugged closely to light source module's one side, and the light of the specific colour of another side emission, after the light passed 96 hole micropore boards of placing on the micropore board heat preservation frame of accuse temperature system, is gathered through optic fibre by image transmission system, finally is converted into image signal by image acquisition system and transmits for mutual and central processing module.
Further, 96 groups of light-emitting sources are distributed in the light source module according to the positions corresponding to 96 holes of the micro-hole plate, each group of light-emitting sources comprises a red light-emitting source, a green light-emitting source and a blue light-emitting source, and a light-homogenizing plate is covered on the top of a light-emitting surface of the light source module.
Further, the temperature control system comprises a heat conducting liquid circulating pump, a light source radiator, a first heat conducting liquid connecting pipe, a second heat conducting liquid connecting pipe, a micro-pore plate heat preservation frame, heating coils and a temperature sensor, wherein the heat conducting liquid circulating pump is connected with the light source radiator, the light source radiator is made of metals with good heat conductivity such as copper, aluminum and silver, a heat conducting liquid flow path is arranged in the light source radiator, the light source module is tightly attached to the bottom surface of the light source radiator, the first heat conducting liquid connecting pipe and the second heat conducting liquid connecting pipe are connected with the light source radiator and the micro-pore plate heat preservation frame to form a heat conducting liquid closed loop, the micro-pore plate heat preservation frame is made of metals with good heat conductivity such as copper, aluminum and silver, through holes are reserved in the positions of the holes on the corresponding 96-pore plate heat preservation frame, the heating coils are encircling and tightly attached to the upper part of the micro-pore plate heat preservation frame, and the temperature sensor is tightly attached to the surface of the micro-pore plate heat preservation frame.
Further, the image transmission system comprises a condensing lens, an optical fiber fixing seat, an optical fiber bundle and an optical fiber bundling device, wherein the condensing lens is arranged in 96 through holes of the micro-hole plate heat preservation frame, the distance between the condensing lens and the section of the optical fiber bundle below is the focal length of the lens, the optical fiber fixing seat is respectively arranged at the bottom of each through hole, the optical fiber bundle consists of 96 optical fibers, one end of the optical fiber bundle is coaxially connected with the optical fiber fixing seat, and the other end of the optical fiber bundle is converged into a bundle at the optical fiber bundling device.
Further, the image acquisition system comprises an optical filter selector, an optical filter, an image acquisition lens and a camera, wherein a plurality of optical filters are uniformly distributed on the optical filter selector in the same diameter, the optical filters can be adjusted to the position coaxial with a light beam gathering device in the image transmission system, the image acquisition lens and the camera are sequentially coaxially installed with the optical fiber gathering device, the optical filter is arranged on the optical fiber gathering device, the camera is arranged above the optical fiber gathering device, and the distance between the optical fiber gathering device and the camera is the focal length of the image acquisition lens.
Further, the interaction and central processing module is provided with a touch screen, the color, brightness and color temperature of the light source module and the temperature maintained by the temperature control system can be modulated, RGB parameters of an image can be extracted, a standard curve is calculated according to parameter values of specific positions where standard substances are placed, parameter values of other positions are substituted into the standard curve for comparison to obtain a detection result, and the image detection result is presented to a user or shared to other intelligent terminals.
The principle of the invention is as follows: some biochemical analysis techniques such as ELISA measurement techniques obtain detection results by quantitatively analyzing the absorbance of a reaction substrate in a microplate. The absorbance can be reflected by the color shade degree of the image in the image technology, and the accuracy and the sensitivity of the image on absorbance reflection can be improved by modulating parameters such as background light color, color temperature and the like and using an optical filter. The principle of the invention is that when biochemical detection steps such as ELISA and the like are completed, a microplate with a reaction substrate is placed at a corresponding position of a microplate reader, and light emitted by a light source according to the modulated parameters is transmitted through the reaction substrate, converged by a lens and then projected onto the section of an optical fiber. The optical fiber can change the optical path of the optical fiber without damage, and the imaging area is reduced by bundling the optical fiber. Finally, the camera acquires an image after the optical fibers are clustered, and the information such as the concentration of the specific substances in the corresponding reaction substrates is analyzed according to the color conditions of the corresponding positions in the image.
Compared with the prior art, the invention has the following advantages and effects:
(1) Compared with an ELISA reader, the invention has higher sensitivity and shorter analysis time, and has more visual and flexible display modes through corresponding software, thereby realizing real-time issuing, long-term storage and network sharing of the diagnosis report.
(2) The invention has small volume, light weight, good portability, simple operation, better meets the POCT requirement, can be more flexibly adapted to various environments, can realize on-site and bedside detection, and is more suitable for basic medical units and household detection to cope with sudden public health events.
(3) The invention has simple structure and lower power consumption, improves the stability and reliability of the instrument, and reduces the purchase and maintenance cost of the instrument.
(4) The invention has wider application field, and can be used for various detection means and projects such as ELISA, spectrophotometry and the like.
Drawings
Fig. 1 is a schematic structural diagram of the present invention, wherein: the device comprises a 1-interaction and central processing module, a 2-camera, a 3-image acquisition lens, a 4-optical filter, a 5-optical filter selector, a 6-optical fiber bundling device, a 7-optical fiber bundle, an 8-optical fiber fixing seat, a 9-temperature sensor, a 10-heating coil, a 11-micropore plate heat preservation frame, a 12-condensing lens, a 13-kit, a 14-first heat conducting liquid connecting pipe, a 15-second heat conducting liquid connecting pipe, a 16-light source module, a 17-light source radiator and a 18-heat conducting liquid circulating pump;
fig. 2 is a schematic structural diagram of a light source module according to the present invention, wherein: 19-dodging plate, 20-red light source, 21-green light source, 22-blue light source.
Detailed Description
Embodiments of the present invention are described in further detail below with reference to the attached drawings; the present embodiments are to be considered as illustrative and not restrictive, and the scope of the invention is not to be limited thereto.
Example 1
As shown in fig. 1 and 2, the invention provides a 96-hole microplate reader, which comprises an interaction and central processing module, a light source module, a temperature control system, an image transmission system and an image acquisition system, wherein the interaction and central processing module 1 is provided with a large touch screen, and is connected with the light source module 16, a temperature sensor 9 of the temperature control system, a heating coil 10 and a camera 2 of the image acquisition system through wires, so that a feedback result of the temperature sensor 9 can be obtained, parameters of the light source module 16 and the heating coil 10 can be modulated, and meanwhile, the acquired image result can be processed or shared with other intelligent terminals; the light source module 16 is provided with 96 groups of light emitting sources according to the corresponding positions of 96 holes of the micro-hole plate, each group of light emitting sources comprises a red light emitting source 20, a green light emitting source 21 and a blue light emitting source 22, the top of the light emitting surface is covered with a light homogenizing plate, one surface can emit light rays with specific colors according to parameters set by the interaction and the central processing module, and the other surface is tightly attached to a light source radiator 17 of the temperature control system to emit heat generated by work to maintain a stable working state; the temperature control system comprises a heat conducting liquid circulating pump 18, a light source radiator 17, a first heat conducting liquid connecting pipe 14, a second heat conducting liquid connecting pipe 15, a micro-pore plate heat preservation frame 11, a heating coil 10 and a temperature sensor 9, wherein the heat conducting liquid circulating pump 18 is connected with the light source radiator 17 to drive heat conducting liquid to transfer heat generated by a light source module 16 to the micro-pore plate heat preservation frame 11 for heat preservation of the micro-pore plate 13 through the internal circulation of a closed loop formed by the internal flow path of the light source radiator 17, the first heat conducting liquid connecting pipe 14, the internal flow path of the micro-pore plate heat preservation frame 11 and the second heat conducting liquid connecting pipe 15, the micro-pore plate 13 is placed on the micro-pore plate heat preservation frame 11, through holes are reserved at corresponding positions of the holes, the heating coil 10 is wound on the upper part of the micro-pore plate heat preservation frame 11 for preheating and rapid temperature rising when the temperature is insufficient, and the temperature sensor 9 is tightly clung to the surface of the micro-pore plate heat preservation frame 11 for feeding back the current maintained temperature; the image transmission system comprises a condensing lens 12, an optical fiber fixing seat 8, an optical fiber bundle 7 and an optical fiber bundling device 6, wherein light rays emitted by a light source module 16 pass through a micro-pore plate 13 arranged on a micro-pore plate heat preservation frame 11, are focused to the optical fiber bundle 7 consisting of 96 optical fibers on the optical fiber fixing seat 8 arranged at the bottom of the micro-pore plate heat preservation frame 11 by the condensing lens 12 in 96 through holes of the micro-pore plate heat preservation frame 11, and are converged at the optical fiber bundling device 6 by an optical fiber bundle 7 adjusting light path; the image acquisition system comprises an optical filter selector 5, an optical filter 4, an image acquisition lens 3 and a camera 2, wherein an image formed by converging optical fibers 7 on an optical fiber bundling device 6 passes through the optical filter 4 selected on the optical filter selector 4, the imaging range is adjusted by the image acquisition lens 3, and finally the image is captured by the camera 2 and converted into an image signal to be transmitted to the interaction and central processing module 1.
Example 2
Creatinine in urine was detected based on a 96-well microplate reader as shown in fig. 1, as follows:
(1) The temperature of the temperature control system is set to 37 ℃ by the 96-hole microplate reader, the optical filter is selected to be 510nm, the light source module is set to be green light, preheating is started, and background light calibration is completed.
(2) The microwell plate was prepared with 9 wells each having a volume of 150. Mu.L, and concentrations of 0. Mu.M, 2.5. Mu.M, 5. Mu.M, 10. Mu.M, 12.5. Mu.M, 20. Mu.M, 25. Mu.M, 50. Mu.M, and 100. Mu.M in creatinine solution.
(3) 50 μl of urine sample to be tested is diluted 200 times and 150 μl is placed in the 10 th well of the microplate.
(4) To each well was added 50. Mu.L of 25mM picric acid solution and 50. Mu.L of 0.75M sodium hydroxide solution.
(5) The microplate was placed in a reader and the mixed solution was allowed to react at 37℃for 10 minutes.
(6) And recording the image at the moment by using a reader, analyzing the image of each hole to obtain data of each hole, generating a standard curve by using the data of the previous nine holes and substituting the standard curve into the data of the 10 th hole, and finally obtaining the concentration of creatinine in urine.
The invention simplifies the operation flow of detection and improves the convenience of instrument use; meanwhile, the detection speed is improved, and the states of all holes of the micro-pore plate can be synchronously observed in real time; it is also possible to store a large amount of verification data and to review or share this data with other intelligent terminals.
The above-described embodiments are intended to illustrate the present invention, not to limit it, and any modifications and variations made thereto are within the spirit of the invention and the scope of the appended claims.
Claims (5)
1. A 96-well microplate reader, characterized in that: the reader comprises an interaction and central processing module, a light source module, a temperature control system, an image transmission system and an image acquisition system, wherein the interaction and central processing module is connected with the light source module, the temperature control system and the image acquisition system through wires at the same time, one surface of the light source module is tightly attached to a light source radiator of the temperature control system, the other surface of the light source module emits light with a specific color, and after the light passes through a 96-hole micro-pore plate placed on a micro-pore plate heat preservation frame of the temperature control system, the light is gathered by the image transmission system through optical fibers and finally converted into an image signal by the image acquisition system to be transmitted to the interaction and central processing module;
the temperature control system comprises a heat conducting liquid circulating pump, a light source radiator, a first heat conducting liquid connecting pipe, a second heat conducting liquid connecting pipe, a micro-pore plate heat preservation frame, a heating coil and a temperature sensor, wherein the heat conducting liquid circulating pump is connected with the light source radiator, the light source radiator is made of metals with good heat conductivity such as copper, aluminum and silver, a heat conducting liquid flow path is arranged in the light source radiator, the bottom surface of the light source radiator is tightly attached to a light source module, the first heat conducting liquid connecting pipe and the second heat conducting liquid connecting pipe are connected with the light source radiator and the micro-pore plate heat preservation frame to form a heat conducting liquid closed loop, the micro-pore plate heat preservation frame is made of metals with good heat conductivity such as copper, aluminum and silver, through holes are reserved in positions corresponding to all holes on the 96-pore plate heat preservation frame, the heating coil is tightly attached to the upper part of the micro-pore plate heat preservation frame in a surrounding mode, and the temperature sensor is tightly attached to the surface of the micro-pore plate heat preservation frame.
2. The 96-well microplate reader of claim 1, wherein: the light source module is provided with 96 groups of light emitting sources distributed according to the corresponding positions of 96 holes of the micro-hole plate, each group of light emitting sources comprises a red light emitting source, a green light emitting source and a blue light emitting source, and the top of the light emitting surface of the light source module is covered with a light homogenizing plate.
3. The 96-well microplate reader of claim 1, wherein: the image transmission system comprises a condensing lens, an optical fiber fixing seat, an optical fiber bundle and an optical fiber bundling device, wherein the condensing lens is arranged in 96 through holes of a micro-hole plate heat preservation frame, the distance between the condensing lens and the section of the optical fiber bundle below is the focal length of the lens, the optical fiber fixing seat is respectively arranged at the bottom of each through hole, the optical fiber bundle consists of 96 optical fibers, one end of the optical fiber bundle is coaxially connected with the optical fiber fixing seat, and the other end of the optical fiber bundle is converged into one bundle at the optical fiber bundling device.
4. The 96-well microplate reader of claim 1, wherein: the image acquisition system comprises an optical filter selector, optical filters, an image acquisition lens and a camera, wherein a plurality of optical filters are uniformly arranged on the optical filter selector in the same diameter, the optical filters can be adjusted to the position coaxial with a light beam gathering device in the image transmission system, the image acquisition lens and the camera are sequentially and coaxially arranged with the optical fiber gathering device, the optical filters are arranged on the optical fiber gathering device, the camera is arranged above the optical fiber gathering device, and the distance between the optical fiber gathering device and the camera is the focal length of the image acquisition lens.
5. The 96-well microplate reader of claim 1, wherein: the interaction and central processing module is provided with a touch screen, and can modulate the color, brightness and color temperature of the light source module and the temperature maintained by the temperature control system; the RGB parameters of the image can be extracted, a standard curve is calculated according to the parameter values of the specific positions where the standard substance is placed, the parameter values of other positions are substituted into the standard curve for comparison to obtain a detection result, and the image detection result is presented to a user or shared with other intelligent terminals.
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| CN202110610917.3A CN113358650B (en) | 2021-06-01 | 2021-06-01 | 96 hole micropore board reader |
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