CN112858246B - Micro-droplet chip analyzer containing multi-light path assembly - Google Patents
Micro-droplet chip analyzer containing multi-light path assembly Download PDFInfo
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- 238000001514 detection method Methods 0.000 abstract description 15
- 239000007788 liquid Substances 0.000 abstract description 10
- 230000005284 excitation Effects 0.000 description 25
- 239000007850 fluorescent dye Substances 0.000 description 17
- 238000004458 analytical method Methods 0.000 description 8
- 238000000034 method Methods 0.000 description 6
- 239000013307 optical fiber Substances 0.000 description 6
- 238000010586 diagram Methods 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 239000000975 dye Substances 0.000 description 3
- 230000004907 flux Effects 0.000 description 3
- 230000033001 locomotion Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 2
- 238000007847 digital PCR Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
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- 230000001351 cycling effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000000695 excitation spectrum Methods 0.000 description 1
- 238000002284 excitation--emission spectrum Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000003754 machining Methods 0.000 description 1
- 238000001921 nucleic acid quantification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
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- 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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- 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
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Abstract
The invention provides a micro-droplet chip analyzer with a multi-light-path assembly, which comprises a first multi-light-path assembly and a second multi-light-path assembly, wherein the first multi-light-path assembly and the second multi-light-path assembly respectively comprise a plurality of LED light source assemblies, the LED light source assemblies are used for emitting exciting light with preset wavelengths, the preset wavelengths of the exciting light emitted by the LED light source assemblies in the first multi-light-path assembly and the preset wavelengths of the exciting light emitted by the LED light source assemblies in the second multi-light-path assembly are different from each other and are adjacent at intervals. According to the invention, the preset wavelengths of the exciting light emitted by the LED light source assemblies respectively arranged on the first multi-light-path assembly and the second multi-light-path assembly are different from each other and are adjacent at intervals, so that the difference between the preset wavelengths of the exciting light emitted by the same multi-light-path assembly is larger, the problem of interference between adjacent preset wavelength signals can be better solved, the single detection index of the detection liquid is obviously increased, and the detection efficiency is improved.
Description
Technical Field
The invention belongs to the technical field of digital PCR analyzers, and particularly relates to a micro-droplet chip analyzer with a multi-light-path component.
Background
Digital PCR is a recent quantitative technique, which is an absolute quantitative method for nucleic acid quantification based on a single-molecule PCR method for counting. The method mainly adopts a micro-fluidic or micro-droplet method to disperse a large amount of diluted nucleic acid solution into micro-reactors or micro-droplets of a chip, wherein the number of nucleic acid templates in each reactor is less than or equal to 1. Thus, after PCR cycling, the microdroplets are irradiated with light of a particular wavelength, a reactor with a nucleic acid molecule template will give a particular fluorescent signal, and a reactor without a template will not give a particular fluorescent signal. Based on the relative proportions and the volume of the reactor, the nucleic acid concentration of the original solution can be deduced. In a micro-droplet chip analyzer, a light path device is an important device for irradiating a micro-droplet with excitation light of a specific wavelength and causing a fluorophore in the micro-droplet to emit fluorescence of a specific wavelength. Because fluorescence signals with adjacent wavelengths are easy to interfere with each other, and the signal analysis precision is affected, most of the traditional micro droplet chip analyzers adopt 2-optical path devices and 3-optical path devices. With the development of science and technology, higher requirements are put forward for the detection speed, efficiency, flux and the like of the micro droplet chip analyzer in medical institutions, scientific research institutions and the like, and new challenges are put forward for the optical path device of the micro droplet chip analyzer.
Disclosure of Invention
Therefore, the technical problem to be solved by the present invention is to provide a micro droplet chip analyzer including a multi-light path assembly, where the preset wavelengths of the excitation lights respectively emitted by a plurality of LED light source assemblies respectively provided in a first multi-light path assembly and a second multi-light path assembly are different and adjacent to each other at intervals, so as to better solve the interference problem between adjacent preset wavelength signals, significantly increase the single detection index of the detection liquid, and improve the detection efficiency.
In order to solve the above problems, the present invention provides a micro droplet chip analyzer, including a first multi-light-path component and a second multi-light-path component, where the first multi-light-path component and the second multi-light-path component respectively include a plurality of LED light source components, the LED light source components are used to emit excitation light with preset wavelengths, and the preset wavelengths of the excitation light emitted by the plurality of LED light source components in the first multi-light-path component and the preset wavelengths of the excitation light emitted by the plurality of LED light source components in the second multi-light-path component are different from each other and are adjacent to each other at intervals.
Preferably, the micro droplet chip analyzer further includes an optical path supporting component, the optical path supporting component includes a first optical path supporting plate and a second optical path supporting plate which are arranged at an interval, the first optical path supporting plate and the second optical path supporting plate are connected with a first optical path connecting plate and a second optical path connecting plate at an interval, the first optical path connecting plate is connected with the first multi-optical path component, the second optical path connecting plate is connected with the second multi-optical path component, and a space between the first optical path connecting plate and the second optical path connecting plate is used for placing the micro droplet chip.
Preferably, the second optical path connecting plate is slidably connected between the first optical path supporting plate and the second optical path supporting plate along a first direction, and after the position adjustment of the second multi-optical-path component is completed, the position of the second optical path connecting plate can be locked.
Preferably, one side of the first light path connecting plate, which deviates from the second light path connecting plate, is connected with a screw motor, the first multi-light-path assembly is connected with the first light path connecting plate through the screw motor, and the screw motor can adjust the height of the first multi-light-path assembly.
Preferably, the screw rod motor is connected to the first light path connecting plate through a motor connecting piece, and the motor connecting piece can drive the screw rod motor and the first multi-light path assembly to slide along a second direction relative to the first light path connecting plate, wherein the second direction is different from the first direction in surface and is orthogonal to the first direction.
Preferably, the first optical path connecting plate is further connected to a displacement adjusting assembly, and the displacement adjusting assembly is configured to adjust a position of the first multi-optical-path assembly in the second direction.
Preferably, the displacement adjusting assembly comprises a fine adjustment platform, a first positioning plate and a second positioning plate, the fine adjustment platform is sequentially connected with the first positioning plate and the second positioning plate towards one side of the motor connecting piece, one side of the second positioning plate, which is far away from the first positioning plate, is connected with the motor connecting piece, and when the fine adjustment platform runs, the first positioning plate and the second positioning plate can force the motor connecting piece to slide along the second direction; and/or, the opposite two sides of the motor connecting piece are provided with guide pieces, the guide pieces are connected with the first light path connecting plate, and the two guide pieces which are arranged oppositely form a sliding guide channel of the motor connecting piece.
Preferably, the axis of the objective lens included in the first multi-light-path assembly and the axis of the objective lens included in the second multi-light-path assembly are parallel to each other and spaced apart by a predetermined distance while being coplanar with the first direction.
Preferably, the preset distance is d, and d is more than or equal to 50 mu m and less than or equal to 200 mu m.
Preferably, the LED light source assembly includes an LED light source, an LED fixing member, a ball lens, a second plano-convex lens, a light filter, and a lens holder, which are linearly arranged in sequence, wherein the LED fixing member, the ball lens, the second plano-convex lens, and the light filter are located in a mounting hole of the lens holder, and an inner side surface of the LED light source is connected to an outer side surface of the lens holder.
According to the micro-droplet chip analyzer provided by the invention, the preset wavelengths of the exciting light respectively emitted by the LED light source assemblies respectively arranged on the first multi-light-path assembly and the second multi-light-path assembly are different and adjacent at intervals, so that the difference between the preset wavelengths of the exciting light emitted by the same multi-light-path assembly is larger, the problem of interference between adjacent preset wavelength signals can be better solved, the single detection index of the detection liquid is obviously increased, and the detection efficiency is improved.
Drawings
FIG. 1 is a schematic structural diagram of a micro droplet chip analyzer according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of the three-beam optical assembly of FIG. 1;
FIG. 3 is a schematic structural diagram (disassembled) of the LED light source assembly in FIG. 1;
FIG. 4 is a schematic structural diagram of the optical path support assembly of FIG. 1;
FIG. 5 is a schematic view of the multi-optical-path module (three-optical-path module) of FIG. 1 (arrows indicate light transmission paths);
fig. 6 shows excitation spectra and emission spectra of four conventional fluorescent dyes, where the ordinate T% is the transmittance, dimensionless, and represents the percentage of the light flux transmitted through a transparent or translucent body to the incident light flux, and the abscissa is the wavelength.
The reference numerals are represented as:
300. an LED light source assembly; 301. a first LED light source assembly; 302. a second LED light source assembly; 303. a third LED light source assembly; 100. a first multi-light-path component; 200. a second multi-light path assembly; 400. An optical path support assembly; 1. an LED light source; 2. an LED fixture; 3. a ball lens; 4. a second plano-convex lens; 5. an optical filter; 6. a lens holder; 7. a first dichroic mirror; 8. a second dichroic mirror; 9. a first mounting member; 10. a second mount; 11. a third mount; 12. an objective lens; 13. a fourth mount; 14. a third dichroic mirror; 15. a fifth mount; 16. a first plano-convex lens; 17. a biphase chromatic mirror gland bush; 18. A fourth dichroic mirror; 19. an optical fiber connector; 20. a lens connecting member; 21. a first optical path support plate; 22. A second optical path support plate; 23. a second optical path connecting plate; 24. a first optical path connecting plate; 25. fine tuning the platform; 26. a first positioning plate; 27. a second positioning plate; 28. a motor connector; 29. a guide member; 30. a screw motor; 31. micro-droplet chips.
Detailed Description
Referring to fig. 1 to 6 in combination, according to an embodiment of the present invention, a droplet chip analyzer including a multi-light-path assembly is provided, which includes a first multi-light-path assembly 100 and a second multi-light-path assembly 200, wherein the first multi-light-path assembly 100 and the second multi-light-path assembly 200 respectively include a plurality of LED light source assemblies 300 therein, the LED light source assemblies 300 are configured to emit excitation light with a predetermined wavelength, and the predetermined wavelength of the excitation light emitted by each of the plurality of LED light source assemblies 300 in the first multi-light-path assembly 100 (the predetermined wavelength is selected according to a specific detection target) and the predetermined wavelength of the excitation light emitted by each of the plurality of LED light source assemblies 300 in the second multi-light-path assembly 200 are different from each other and are adjacent to each other at intervals. In the technical scheme, the preset wavelengths of the excitation lights respectively emitted by the plurality of LED light source assemblies 300 respectively arranged in the first multi-light-path assembly 100 and the second multi-light-path assembly 200 are different from each other and are adjacent at intervals, and the excitation lights with adjacent wavelengths (excitation light and emission light wavelength) can be respectively arranged in the two light-path assemblies, so that the two light paths can be ensured not to be detected on the same liquid drop at any moment, the difference between the preset wavelengths of the excitation lights emitted in the same multi-light-path assembly is large, the interference problem between adjacent preset wavelength signals can be better solved, the single detection index of the detection liquid is remarkably increased, the most efficient wave band is adopted for exciting to obtain the strongest signal value aiming at each fluorescent dye, and the detection efficiency is improved.
Taking the first multi-light-path assembly 100 and the second multi-light-path assembly 200 as three light-path assemblies as an example, each of the three light-path assemblies includes three LED light source assemblies 300, at this time, when the first multi-light-path assembly 100 and the second multi-light-path assembly 200 are simultaneously used, a six-light-path device of the analyzer is formed at this time, the six LED light source assemblies 300 respectively emit laser light with corresponding preset wavelengths, that is, excitation light with six wavelengths, and the six excitation light is set to be a (corresponding to the fluorescent dye a), B (corresponding to the fluorescent dye B), C (corresponding to the fluorescent dye C), D (corresponding to the fluorescent dye D), E (corresponding to the fluorescent dye E), and F (corresponding to the fluorescent dye F) according to the serial numbers from small to large, the wavelength of the laser light is set according to the selection of the fluorescent dye, and according to the actual situation, the excitation wavelengths of the selected six fluorescent dyes are not distributed in an arithmetic progression from 480nm to 700nm, the excitation wavelengths of some fluorescent dyes are very close (such as fluorescent dyes three and four in figure 6), the excitation wavelengths of some fluorescent dyes are greatly different (such as fluorescent dyes one and two in figure 6), if the six-optical-path device is made into a whole, six excitation lights simultaneously excite the six fluorescent dyes, the dye A can be excited by the laser one (excitation light wavelength a), meanwhile, the dye A can be excited by the laser two (excitation light wavelength B), the dye C only has the excitation effect of the laser three (excitation light wavelength C), the emitted fluorescent signals are not uniform in strength, and the signals need to be split by a dichroscope in the subsequent transmission and analysis processes, however, the existing dichroic mirror technology is limited, and a plurality of lights with close wavelengths cannot be effectively split, so that the light intensity of a certain light is weakened, or two lights enter the same photomultiplier, and thus the interference problem of adjacent light signals is caused, and finally the obtained signal cannot reflect the actual situation, but in the present invention, an integral six-light-path device is respectively a first multi-light-path component 100 and a second multi-light-path component 200, taking the first multi-light-path component 100 as an example, the laser serial numbers of the light paths of the first multi-light-path component 100 are one, three, five, and the serial numbers of the target fluorescent dyes to be excited are a, C, and E, in the subsequent signal analysis, only the possible lights of the excited fluorescent dyes B, D, and F need to be filtered, correspondingly, the laser serial numbers of the light paths of the second multi-light-path component 200 are B, D, and F, and the signal analysis is the same as the light paths of the first multi-light-path component 100, and thus the six-light-path forms of the three light paths respectively arranged are integrated with respect to the six-light paths, and thus the signal interference problem is better solved.
In one embodiment, the micro droplet chip analyzer further includes an optical path supporting assembly 400, the optical path supporting assembly 400 includes a first optical path supporting plate 21 and a second optical path supporting plate 22 disposed at an interval, a first optical path connecting plate 24 and a second optical path connecting plate 23 are connected between the first optical path supporting plate 21 and the second optical path supporting plate 22 at an interval, the first optical path connecting plate 24 is connected to the first multi-optical-path assembly 100, the second optical path connecting plate 23 is connected to the second multi-optical-path assembly 200, and a space between the first optical path connecting plate 24 and the second optical path connecting plate 23 is used for placing the micro droplet chip 31. In this embodiment, the first multi-optical-path module 100 and the second multi-optical-path module 200 are respectively disposed on the first optical-path connecting plate 24 and the second optical-path connecting plate 23 in an opposing manner, so that the two multi-optical-path modules can be arranged more compactly, and in a specific implementation, the first multi-optical-path module 100 and the second multi-optical-path module 200 are disposed in an opposing manner, and at this time, the droplet chips 31 are placed in a space therebetween by a robot or manually.
Preferably, the second optical path connecting plate 23 is slidably connected between the first optical path supporting plate 21 and the second optical path supporting plate 22 along a first direction, and after the position adjustment of the second multi-optical-path assembly 200 is completed, the position of the second optical path connecting plate 23 can be locked, the second optical path connecting plate 23 is designed to be slidably connected along the first direction, so that the machining size error caused by the second optical path connecting plate 23 adopting a fixed connection mode can be effectively avoided, and the position of the second optical path connecting plate is more accurate. The aforementioned first direction may specifically be a direction (inward or outward from the paper) which is horizontal and perpendicular to the paper in the use orientation as shown in fig. 1.
Further, one side of the first optical path connecting plate 24 departing from the second optical path connecting plate 23 is connected to a lead screw motor 30, the first multi-optical-path component 100 is connected to the first optical path connecting plate 24 through the lead screw motor 30, and the lead screw motor 30 can adjust the height of the first multi-optical-path component 100, specifically, the first multi-optical-path component 100 is fixedly connected to a mover of the lead screw motor 30, and when the lead screw motor 30 operates, the mover drives the first multi-optical-path component 100 to perform a reciprocating motion (for example, to be raised or lowered, specifically according to the rotating direction of the lead screw motor 30).
Further, the screw rod motor 30 is connected to the first optical path connecting plate 24 through the motor connecting piece 28, and the motor connecting piece 28 can drive the screw rod motor 30 and the first multi-optical-path assembly 100 is opposite to the first optical path connecting plate 24 slides along the second direction, the second direction is different from the first direction and orthogonal, at this time, the height and the horizontal position of the first multi-optical-path assembly 100 can be flexibly adjusted, and the second direction is different from the first direction and orthogonal, so that the height adjustment effect of the screw rod motor 30 is compounded, and the focusing adjustment of the objective lenses 12 in the two optical-path assemblies 100 is more accurate and convenient. It can be understood that, a displacement adjusting assembly is further connected to the first optical path connecting plate 24, the displacement adjusting assembly is used for adjusting the position of the first multi-optical-path assembly 100 in the second direction, specifically, the displacement adjusting assembly includes a fine tuning platform 25, a first positioning plate 26, and a second positioning plate 27, the fine tuning platform 25 is connected to the first positioning plate 26 and the second positioning plate 27 in sequence towards one side of the motor connecting member 28, one side of the second positioning plate 27 away from the first positioning plate 26 is connected to the motor connecting member 28, and when the fine tuning platform 25 operates, the first positioning plate 26 and the second positioning plate 27 can force the motor connecting member 28 to slide in the second direction; and/or, two opposite sides of the motor connector 28 are provided with guide members 29, the guide members 29 are connected with the first light path connecting plate 24, and the two opposite guide members 29 form a sliding guide channel of the motor connector 28. When the horizontal position of the objective lens 12 (i.e., the first direction and the second direction) is determined, the position of the motor connector 28 relative to the first optical path connecting plate 24 is locked, and then the displacement adjusting assembly may be removed.
In some embodiments, the axis of the objective lens 12 in the first multi-optical-path assembly 100 and the axis of the objective lens 12 in the second multi-optical-path assembly 200 are parallel to each other and separated by a predetermined distance, and are coplanar with the first direction, and the predetermined distance is preferably between 50 μm and 200 μm, as verified by the inventor, in this range, the signal intensity is clearly contrasted, and the signal interference resistance is more obvious.
Preferably, the first multi-light-path assembly 100 and the second multi-light-path assembly 200 have the same structure, and it is understood that the other structures are the same except for the LED light source assemblies 300 respectively provided therein. The specific structure of the first multi-optical-path component 100 is described below as a three-optical-path component: the first multi-light-path component 100 includes a first LED light source component 301, a second LED light source component 302, a third LED light source component 303, a first dichroic mirror 7, a second dichroic mirror 8, a third dichroic mirror 14, an objective lens 12, a first plano-convex lens 16, and a fourth dichroic mirror 18, excitation lights respectively emitted by the first LED light source component 301 and the second LED light source component 302 sequentially pass through the first dichroic mirror 7, the second dichroic mirror 8, and the third dichroic mirror 14, the excitation light emitted by the third LED light source component 303 is emitted from the objective lens 12 to micro droplets in a channel of a micro droplet chip 31 after passing through the second dichroic mirror 8 and the third dichroic mirror 14, and fluorescence generated by excitation sequentially passes through the objective lens 12, the third dichroic mirror 14, and the first plano-convex lens 16, enters the optical fiber connector 19 and/or the lens connector 20 at the fourth dichroic mirror 18, the optical fiber connector 19 is connected with optical fibers to transmit corresponding fluorescence signals to electronic components such as photomultiplier tubes, so as to convert the signals into electrical signals, and record the movement of the micro droplets in the micro camera, and display the micro droplets on the micro-camera chip.
Further, the first multi-light-path assembly 100 comprises a structural body, the structural body comprises a first mounting piece 9, a second mounting piece 10, a third mounting piece 11, a fourth mounting piece 13 and a fifth mounting piece 15, the first mounting piece 9, the second mounting piece 10, the third mounting piece 11, the fourth mounting piece 13 and the fifth mounting piece 15 are assembled into a whole, a light path channel is formed inside the first mounting piece 9, the first dichroic mirror 7 is clamped at the connection position of the first mounting piece 9 and the second mounting piece 10, and the second dichroic mirror 8 is clamped at the connection position of the first mounting piece 9 and the third mounting piece 11; the objective lens 12 is connected to the mounting hole of the third mounting part 11; the third dichroic mirror 14 is interposed at a connection part of the third mounting member 11 and the fourth mounting member 13; the first plano-convex lens 16 is clamped at the joint of the fourth mounting piece 13 and the fifth mounting piece 15; the fourth dichroscope 18 is clamped between the dichroscope gland 17 and the fifth mounting part 15; the optical fiber connector 19 is connected to an end of the fifth mounting member 15, and the lens connector 20 is connected to a side of the fifth mounting member 15.
The LED light source assembly 300 includes an LED light source 1, an LED fixing member 2, a spherical lens 3, a second plano-convex lens 4, an optical filter 5, and a lens holder 6, which are linearly arranged in sequence, wherein the LED fixing member 2, the spherical lens 3, the second plano-convex lens 4, and the optical filter 5 are located in a mounting hole of the lens holder 6, and an inner side surface of the LED light source 1 is connected with an outer side surface of the lens holder 6.
The micro-droplet chip analyzer of the invention needs to perform necessary position determination operation during assembly and debugging, and can be specifically performed by adopting the following modes:
referring to fig. 1 and 4 in combination, when the position is determined during initial installation, the six LED light sources 1 are powered on to emit light with 6 different wavelengths; the screw motor 30 acts to lift the first three optical path assembly (i.e. a specific implementation form of the first multi-optical path assembly 100) to the highest position, so as to make room for placing the micro-droplet chip 31; the micro-droplet chip 31 is placed on a specific structure between the first and second multi-optical-path components (i.e. a specific implementation form of the second multi-optical-path component 200) by a manipulator or manually, the specific structure drives the chip to slightly move left and right, up and down (the orientation shown in fig. 1) under algorithm control, so that the micro-droplet in the trench is just at the focus of the objective lens 12 in the second multi-optical-path component, at this time, the micro-droplet can form a clear image on the display screen of the industrial computer, then the lead screw motor 30 acts to lower the first multi-optical-path component to a certain position, the first multi-optical-path component is finally driven to slightly move by the fine tuning platform 25 on the manual fine tuning optical-path supporting component 400, the first positioning plate 26, the second positioning plate 27, the motor connecting piece 28 and the lead screw motor 29, and by observing the imaging of the camera on the display screen of the industrial computer on the first multi-optical-path component, the light emitted by the objective lens on the first light path component is aligned with the groove on the micro-droplet chip 31, then the bolt on the motor connecting piece 28 is fixed, the position of the first light path component is fixed relative to the light path supporting component 400 through the motor connecting piece 28 and the screw rod motor 30, thus the axes of the upper and lower two objective lenses 12 are positioned on the same vertical plane as shown in figure 1, the fine tuning platform 25, the first positioning plate 26, the second positioning plate 27 and the guiding piece 29 are taken as the assembly for assembly and can be removed, the front and back positions of the second light path component are finely tuned through manually fine tuning the first light path connecting plate 23, so that the axes of the upper and lower two objective lenses 12 are positioned on two different planes which are parallel to the paper surface and have the distance of 50-200 μm, the bolt on the first light path connecting plate 23 is fastened, and the position of the second light path component v is fixed, no change is made, and the relative position (horizontal relative position) of the first and second third optical path components in the analyzer is determined, and the height mode completely depends on the up-and-down adjustment of the screw motor 30 (and it can be understood that the detection function after the analyzer does not need to perform the position determination process). And then, subsequent detection operation can be performed, specifically, the screw rod motor 30 continues to perform up-and-down jogging, so that the micro liquid drops in the channel are exactly positioned at the focus of the objective lens 12 in the first three light path component, a controller of the screw rod motor 30 remembers the position at this time, the position can be marked as the focus position of the first three light path component and is used as a position of subsequent repeated movement, the liquid drops in the channel of the micro liquid drop chip 31 move from far to near (parallel to the first direction) in the direction perpendicular to the paper surface, the liquid drops sequentially pass through irradiation of light rays emitted by the first three light path components and the second three light path components, namely the liquid drops are respectively irradiated by 3 light rays with different wavelengths at two positions and are respectively excited to generate 3 fluorescence with different wavelengths, the 2 groups of excited fluorescence respectively pass through the respective three light path components and finally enter 2 groups of optical devices such as photomultiplier tubes along optical fibers to be converted into electric signals, analysis is performed through an algorithm, the 2 groups of 3 light path fluorescence signal analysis results are fitted, and finally 6 light path fluorescence signal analysis results are output. After the droplet analysis is completed, the lead screw motor 30 is operated to lift the first third optical path component to the highest position, and the chip 31 is taken out by a manipulator or manually and placed in a specified recovery device.
It is readily understood by a person skilled in the art that the advantageous ways described above can be freely combined, superimposed without conflict.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention. The above is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several improvements and modifications can be made without departing from the technical principle of the present invention, and these improvements and modifications should also be regarded as the protection scope of the present invention.
Claims (8)
1. The micro-droplet chip analyzer with the multi-light-path component is characterized by comprising a first multi-light-path component (100) and a second multi-light-path component (200), wherein the first multi-light-path component (100) and the second multi-light-path component (200) respectively comprise a plurality of LED light source components (300), the LED light source components (300) are used for emitting exciting light with preset wavelengths, and the preset wavelengths of the exciting light respectively emitted by the LED light source components (300) in the first multi-light-path component (100) and the preset wavelengths of the exciting light respectively emitted by the LED light source components (300) in the second multi-light-path component (200) are different from each other and are adjacent at intervals;
the optical path support assembly (400) comprises a first optical path support plate (21) and a second optical path support plate (22) which are arranged at intervals relatively, a first optical path connecting plate (24) and a second optical path connecting plate (23) are connected between the first optical path support plate (21) and the second optical path support plate (22) at intervals relatively, the first optical path connecting plate (24) is connected with the first multi-optical-path assembly (100), the second optical path connecting plate (23) is connected with the second multi-optical-path assembly (200), and a space between the first optical path connecting plate (24) and the second optical path connecting plate (23) is used for placing a micro-droplet chip (31);
the second optical path connecting plate (23) is slidably connected between the first optical path supporting plate (21) and the second optical path supporting plate (22) in a first direction;
the first multi-optical-path assembly (100) slides along a second direction relative to the first optical-path connecting plate (24), and the second direction is different from and orthogonal to the first direction;
the axial line of the objective lens (12) provided in the first multi-light-path assembly (100) and the axial line of the objective lens (12) provided in the second multi-light-path assembly (200) are parallel to each other and spaced apart by a predetermined distance while being coplanar with the first direction.
2. The micro droplet chip analyzer according to claim 1, wherein the position of the second optical path connection plate (23) can be locked after the position adjustment of the second multi optical path module (200) is finished.
3. The microfluidic chip analyzer according to claim 1, wherein a lead screw motor (30) is connected to a side of the first optical path connecting plate (24) away from the second optical path connecting plate (23), the first multi-optical-path assembly (100) is connected to the first optical path connecting plate (24) through the lead screw motor (30), and the lead screw motor (30) can adjust the height of the first multi-optical-path assembly (100).
4. The microfluidic chip analyzer according to claim 3, wherein the lead screw motor (30) is connected to the first optical path connecting plate (24) through a motor connector (28), and the motor connector (28) can drive the lead screw motor (30) and the first multi-optical-path assembly (100) to slide along the second direction relative to the first optical path connecting plate (24).
5. The microfluidic chip analyzer according to claim 4, wherein a displacement adjusting assembly is further connected to the first optical path connecting plate (24), and the displacement adjusting assembly is used for adjusting the position of the first multi-optical path assembly (100) in the second direction.
6. The micro droplet chip analyzer according to claim 5, wherein the displacement adjusting assembly comprises a fine adjusting platform (25), a first positioning plate (26), and a second positioning plate (27), the fine adjusting platform (25) is connected to the first positioning plate (26) and the second positioning plate (27) in sequence on a side facing the motor connecting member (28), the second positioning plate (27) is connected to the motor connecting member (28) on a side away from the first positioning plate (26), and when the fine adjusting platform (25) operates, the first positioning plate (26) and the second positioning plate (27) can force the motor connecting member (28) to slide along the second direction; and/or guide pieces (29) are arranged on two opposite sides of the motor connecting piece (28), the guide pieces (29) are connected with the first light path connecting plate (24), and the two oppositely arranged guide pieces (29) form a sliding guide channel of the motor connecting piece (28).
7. The micro droplet chip analyzer of claim 1, wherein the predetermined distance is d, d is 50 μm or less and 200 μm or less.
8. The microfluidic chip analyzer according to claim 1, wherein the LED light source assembly (300) comprises an LED light source (1), an LED holder (2), a ball lens (3), a second plano-convex lens (4), a light filter (5) and a lens holder (6) which are linearly arranged in sequence, wherein the LED holder (2), the ball lens (3), the second plano-convex lens (4) and the light filter (5) are located in a mounting hole of the lens holder (6), and an inner side surface of the LED light source (1) is connected with an outer side surface of the lens holder (6).
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| KR100818351B1 (en) * | 2005-10-28 | 2008-04-01 | 굿젠 주식회사 | Multichannel biochip scanner |
| JP5279481B2 (en) * | 2008-12-25 | 2013-09-04 | 株式会社日立ハイテクノロジーズ | Nucleic acid analyzer |
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| CN208028908U (en) * | 2018-05-17 | 2018-10-30 | 大连优迅科技有限公司 | Device for reducing multi-wavelength crosstalk and optical path system |
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| CN211227169U (en) * | 2019-12-04 | 2020-08-11 | 苏州东胜兴业科学仪器有限公司 | Multichannel fluorescence quantitative PCR detection system |
| CN214472777U (en) * | 2021-03-14 | 2021-10-22 | 新羿制造科技（北京）有限公司 | Micro-droplet chip analyzer with linearly arranged LED light sources |
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