CN211627376U - Planar miniature multi-channel fluorescence detection optical system - Google Patents
Planar miniature multi-channel fluorescence detection optical system Download PDFInfo
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- CN211627376U CN211627376U CN202020318062.8U CN202020318062U CN211627376U CN 211627376 U CN211627376 U CN 211627376U CN 202020318062 U CN202020318062 U CN 202020318062U CN 211627376 U CN211627376 U CN 211627376U
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Abstract
The utility model relates to a miniature multichannel fluorescence detection optical system of plane formula, include: a planar excitation light assembly and a planar lighting assembly; the reaction tanks are respectively connected to the two components through optical fibers; the planar excitation light assembly includes: the single LED light source light penetrates through the dichroic mirror as single-channel light after being filtered and collimated and/or is focused to the optical fiber after being reflected by the dichroic mirror; single-channel light rays in different directions are output as a group of light ray bundles in the same emergent direction after passing through a first-stage dichroic mirror; the light ray bundles in different directions are output as a group of multi-channel light ray bundle groups in the same emergent direction after passing through a second-stage dichroic mirror; planar daylighting subassembly includes: the fluorescence reaction penetrates through the dichroic mirror after passing through the cylindrical lens and/or is separated into a plurality of single-channel fluorescence in different directions after being reflected by the dichroic mirror, and the single-channel fluorescence is transmitted to a photosensitive surface of the photodiode after being focused and filtered; the fluorescence in the single direction is output as fluorescence beams in different emergent directions after passing through the dichroic mirror of the second stage; each group of fluorescent beams is output as single-channel fluorescent light in different emergent directions after passing through the first-stage dichroic mirror.
Description
Technical Field
The utility model relates to a multichannel fluorescence detects technical field, especially relates to a miniature multichannel fluorescence detection optical system of plane formula.
Background
The fluorescence detection is a natural luminescence reaction, and can detect human cells, bacteria, mould and food residues by reacting luciferase with ATP, and obtain a reaction result within 15 seconds. The fluorescence illuminance is generally measured by a light-sensing device and is represented in digital form.
At present, the most widely used fluorescence detection is the fluorescence quantitative PCR technology, which is a method of adding a fluorescent group into a PCR reaction system, realizing real-time monitoring of the whole process of PCR through continuous accumulation of fluorescence signals, and then carrying out quantitative analysis on an unknown template through a standard curve.
In general, in fluorescence detection technology, a corresponding optical system needs to be designed to ensure a stable state of generated light and fluorescence. The conventional fluorescence detection optical system can only use one optical system to detect one channel, or use a mechanical turntable to use a plurality of optical systems to meet the requirement of detecting a plurality of channels. This leads to problems of large size, heavy weight, and difficult operation of the optical system.
SUMMERY OF THE UTILITY MODEL
The technical scheme of the utility model is that: a planar micro multi-channel fluorescence detection optical system can be applied to fluorescence quantitative PCR, portable PCR, rapid PCR and the like, and solves the problems that the traditional fluorescence detection optical system is large in size, cannot be placed in equipment in a three-dimensional mode, and optical components are not installed well. Meanwhile, the problems that a mechanical turntable is required to be used in a traditional multi-channel fluorescence detection optical system and the like are solved.
According to the scheme, N light beams are emitted from the same channel, coupled into the emitting optical fiber and transmitted to the reaction tank, after the excitation process is completed, fluorescence emitted by a detected object is transmitted to the planar lighting system through the receiving optical fiber, the N light beams are emitted from the N channels respectively, and finally signals are collected through the N photodiodes respectively.
Specifically, the planar micro multi-channel fluorescence detection optical system includes: a plane type exciting light component, a reaction cell and a plane type lighting component. Incident light formed by the planar excitation light assembly is transmitted to a sample to be detected through the incident optical fiber, and fluorescence generated by exciting the sample to be detected by the incident light is transmitted to the photodiode through the planar lighting assembly.
The planar excitation light assembly includes: monochromatic LED lamp, filter tube piece, collimating lens, first order dichroic mirror, second order dichroic mirror, first focusing lens, second focusing lens.
The basic light path is: the single LED light source light penetrates the dichroic mirror as single-channel light after being filtered and collimated and/or is focused to the optical fiber after being reflected by the dichroic mirror.
Because the dichroic mirror is characterized by: almost completely transmitting light of certain wavelengths and almost completely reflecting light of other wavelengths. At a given wavelength, both sides of the dichroic mirror are transmissive or both sides are reflective. Therefore, according to the band selection function of the dichroic mirror in the present application, two output directions of the dichroic mirror are defined as: the transmission direction and the reflection direction. The direction of the light path penetrating the dichroic mirror within the specified wavelength is the penetration direction, and the direction of the light path radiated by the dichroic mirror within the specified wavelength is the reflection direction.
Specifically, in the present embodiment, incident light rays in both the transmission direction and the reflection direction have an incident angle of 45 °. Therefore, two incident light paths on the same dichroic mirror are arranged in a vertical orientation as viewed in a plan view of the optical system. However, based on the above-described principle of arrangement, it is not excluded that there is a differential arrangement of other same principles that exists because the dichroic mirrors differ in size and surface slope.
Based on the arrangement method of the dichroic mirrors, in the planar excitation light assembly, single-channel light rays in different directions pass through the first-stage dichroic mirror and are output as a group of light ray bundles in the same emergent direction; and the light ray bundles in different directions are output as a group of multi-channel light ray bundle groups in the same emergent direction after passing through the second-stage dichroic mirror.
In this scheme, the direction of penetrating of every first order dichroic mirror corresponds a single channel light, and its direction of reflection also corresponds a single channel light. That is, two single channel light rays injected into the same first-stage dichroic mirror form a group, and the group of single channel light rays are selected as a light ray bundle of a specified waveband after passing through the first-stage dichroic mirror.
Each first-stage dichroic mirror can output a group of light ray beams with specified wave bands, so that when two light ray beams are used as a group of incident light of the second-stage dichroic mirror, the incident light correspondingly penetrates into the penetrating direction and the reflecting direction of the second-stage dichroic mirror, and is selectively output as a multi-channel light ray beam group with a single emergent direction through secondary wave bands.
Theoretically, the optical system can be expanded in a tree shape, and the arrangement of optical elements can be correspondingly designed only by inputting and outputting light wave bands.
In the scheme, the multi-channel light beam group output by the second-stage dichroic mirror passes through the first focusing lens and the second focusing lens again and then enters the reaction tank through the incident optical fiber.
The reaction tank is a placing area for detecting a sample, specifically, a plurality of grooves for placing sample containers are arranged on the reaction tank, and the optical fibers penetrate through the reaction tank body and then abut against the side walls of the sample containers. Correspondingly, an emergent optical fiber for receiving the fluorescence reaction is also connected to the reaction cell, and the emergent optical fiber is also abutted to the side wall of the sample container. The corresponding emergent optical fiber and incident optical fiber of the same sample container are opposite in corresponding relation and are optimally positioned at the same liquid level height. The outgoing optical fiber is connected to the planar lighting assembly and conducts the detection equipment through the planar lighting assembly.
Planar daylighting subassembly includes: the device comprises a cylindrical lens, a second-stage dichroic mirror, a first-stage dichroic mirror, a focusing lens and an optical filter.
The basic light path is: the fluorescence reaction excited by the sample penetrates through the dichroic mirror after passing through the cylindrical lens and/or is separated into a plurality of single-channel fluorescence in different directions after being reflected by the dichroic mirror, and the single-channel fluorescence is transmitted to the photosensitive surface of the photodiode after being focused and filtered.
The cylindrical lens is adopted for collimation of light beams, so that a plurality of planar excitation light systems are matched with one planar lighting system to detect a plurality of detected objects.
The second-stage dichroic mirror and the first-stage dichroic mirror in the planar lighting assembly are arranged at an angle of 45 degrees, so that two emergent light paths on the same dichroic mirror are arranged in a vertical direction when viewed from a plane of the optical system. However, based on the above-described principle of arrangement, it is not excluded that there is a differential arrangement of other same principles that exists because the dichroic mirrors differ in size and surface slope.
In the scheme, after the fluorescence reaction light in a single direction passes through the second-stage dichroic mirror, the penetrating direction of the second-stage dichroic mirror corresponds to one emergent fluorescent beam, and the reflecting direction of the second-stage dichroic mirror also corresponds to one emergent fluorescent beam. That is, the first-stage dichroic mirror selectively outputs two fluorescence beams of a specified wavelength band.
Based on the characteristics of the dichroic mirror, each group of fluorescent beams passes through the first-stage dichroic mirror and then is output as single-channel fluorescent light in two different emergent directions. Finally, the single-channel fluorescence sequentially passes through the third lens and the optical filter and then reaches the photodiode.
After the arrangement of the optical path system, the fluorescence detection can be carried out on the detected sample. However, the system may vary in placement depending on the measured properties of the sample. For example, if it is necessary to detect N characteristics of a plurality of objects to be detected, it is only necessary to place a plurality of reaction cells, each reaction cell corresponds to one planar excitation light system, and the plurality of reaction cells only need to be matched with one planar lighting system.
The planar excitation light assembly and the planar lighting assembly on the whole are designed and manufactured, and the structural size is reasonably designed to be 40-10 mm, so that the optical system is small in size, thin in thickness and convenient to assemble and install.
The utility model has the advantages that: the N light beams are emitted from the same channel, coupled into a transmitting optical fiber and transmitted to a reaction pool, after the excitation process is completed, fluorescence emitted by the detected object is transmitted to a planar lighting system through a receiving optical fiber, the N light beams are respectively emitted from the N channels, and finally signals are respectively collected through N photodiodes.
Drawings
The invention will be further described with reference to the following drawings and examples:
FIG. 1 is a technical schematic diagram of a planar micro multi-channel fluorescence detection optical system;
the various references in the drawings are: single color LEDs (1-1, 1-2, 1-3, 1-4); excitation light filters (2-1, 2-2, 2-3, 2-4); collimating lenses (3-1, 3-2, 3-3, 3-4); excitation light dichroic mirrors (4-1, 4-2, 4-3); excitation light focusing lenses (5-1, 5-2); an incident optical fiber (6-1); an exit optical fiber (6-2); a reaction tank (7); a cylindrical lens (8); daylighting dichroic mirrors (9-1, 9-2, 9-3); a lighting focusing lens (10-1, 10-2, 10-3, 10-4); a lighting filter (11-1, 11-2, 11-3, 11-4); photoelectric sensors (12-1, 12-2, 12-3, 12-4).
Detailed Description
Example 1
As shown in fig. 1, the planar excitation light assembly includes: the device comprises monochromatic LEDs (1-1, 1-2, 1-3 and 1-4), an excitation light filter (2-1, 2-2, 2-3 and 2-4), a collimating lens (3-1, 3-2, 3-3 and 3-4), an excitation light dichroic mirror (4-1, 4-2 and 4-3) and an excitation light focusing lens (5-1 and 5-2).
As shown in fig. 1, the planar lighting assembly comprises: the photoelectric sensor comprises a cylindrical lens (8), lighting dichroic mirrors (9-1, 9-2 and 9-3), lighting focusing lenses (10-1, 10-2, 10-3 and 10-4), lighting optical filters (11-1, 11-2, 11-3 and 11-4) and photoelectric sensors (12-1, 12-2, 12-3 and 12-4).
As shown in figure 1, a reaction cell (7) is arranged between the planar excitation light assembly and the planar lighting assembly, the reaction cell (7) is butted with the planar excitation light assembly through an incident optical fiber (6-1), and the reaction cell (7) is butted with the planar lighting assembly through an emergent optical fiber (6-2).
The working process is as follows:
the four monochromatic LED lamps emit 4 light beams, stray light is filtered out through the excitation light filters, the light beams are collimated through the collimating lenses, the light beams are combined through the excitation light dichroic mirror, the light beams are focused and coupled through the excitation light focusing lens and enter the incident optical fiber (6-1), the light beams enter the reaction tank (7) to complete the fluorescence excitation process, a plurality of fluorescence beams radiated by a detected object are transmitted to the cylindrical lens (8) through the emergent optical fiber (6-2) to be collimated, the four light beams are split through the lighting dichroic mirror and are focused through the lighting light focusing lens, and finally the light beams irradiate the surface of the photoelectric sensor to collect optical signals after the stray light beams are filtered through the lighting light filters, so that the flow of the whole optical system is completed.
Compared with the traditional fluorescence detection optical system:
1. the scheme adopts a planar mode to place optical components, and the volume of the planar system is only 40 × 10mm, so that the optical system is small in volume, light and thin and convenient to assemble and install.
2. The LED is used as a light source and the photodiode is used as a detector, so that the cost and the system space are greatly reduced compared with the traditional laser and camera, and the LED-based optical system is convenient to use on portable equipment and various micro spaces with requirements on the volume of the optical system.
3. N monochromatic LEDs are adopted, and the filters are used for filtering out stray light, so that the good unicity of the wavelength of the light source can be ensured.
4. The optical fiber is used as an optical transmission medium for connecting the fluorescence detection system and the reaction cell, so that the space and the cost can be well saved.
Example 2:
the scheme comprises excitation light dichroic mirrors (4-1, 4-2 and 4-3) and lighting dichroic mirrors (9-1, 9-2 and 9-3).
Specifically, the transmission and reflection bands of each dichroic mirror are:
the specification of the excitation light dichroic mirror (4-1) is: reflection at 400nm-485nm and transmission at 500nm-700 nm;
the specification of the excitation light dichroic mirror (4-2) is: reflection at 500nm-590nm and transmission at 600nm-700 nm;
the specification of the excitation light dichroic mirror (4-3) is: reflection at 400nm-550nm and transmission at 560nm-700 nm;
the specification of the lighting dichroic mirror (9-1) is as follows: transmission at 450nm-580nm and reflection at 600nm-750 nm;
the specification of the lighting dichroic mirror (9-2) is as follows: reflection at 500nm-630nm and transmission at 650nm-700 nm;
the specification of the lighting dichroic mirror (9-3) is as follows: 400-535nm reflection and 550-700 nm transmission;
based on the specific parameters of the dichroic mirror, the planar micro multi-channel fluorescence detection optical system specifically comprises the following components:
as shown in figure 1, a monochromatic LED (1-1) of a first channel on a planar excitation light assembly emits light, stray light is filtered by an excitation light filter (2-1), the light is collimated by an excitation light collimating lens (3-1), then the light is reflected to the surface of an excitation light dichroic mirror (4-3) placed at 45 degrees from an excitation light dichroic mirror (4-1) placed at 45 degrees, the light is reflected by the surface of the excitation light dichroic mirror (4-3) and then reaches excitation light focusing lenses (5-1 and 5-2) to be coupled and enter an incident optical fiber (6-1), and the incident optical fiber (6-1) is connected into a reaction tank (7).
Light beams excited by a sample in the reaction cell (7) are transmitted to the cylindrical lens (8) through the emergent optical fiber (6-2), the collimated light beams are transmitted to the 45-degree lighting dichroic mirror (9-3) from the 45-degree lighting dichroic mirror (9-1), the light beams are reflected by the lighting dichroic mirror (9-3), focused by the lighting focusing lens (10-1), transmitted through the lighting optical filter (11-1) and finally incident to the surface of the photoelectric sensor (12-1) of the first channel.
As shown in fig. 1, the monochromatic LED (1-2) of the second channel of the planar excitation light assembly emits light, the stray light is filtered by the excitation light filter (2-2), and the light beam is collimated by the excitation light collimating lens (3-2), and then the light beam is transmitted from the excitation light dichroic mirror (4-1) placed at 45 degrees to the surface of the excitation light dichroic mirror (4-3) placed at 45 degrees and is reflected. The reflected light enters an incident optical fiber (6-1) after being coupled by excitation light focusing lenses (5-1 and 5-2), and the incident optical fiber (6-1) is connected into a reaction cell (7).
Light beams excited by a sample in the reaction cell (7) are transmitted to the cylindrical lens (8) through the emergent optical fiber (6-2), the collimated light beams are transmitted to the 45-degree lighting dichroic mirror (9-3) from the 45-degree lighting dichroic mirror (9-1), the light beams are transmitted through the lighting dichroic mirror (9-3), then are focused through the lighting focusing lens (10-2), then penetrate through the lighting optical filter (11-2), and finally enter the surface of the photoelectric sensor (12-2) of the second channel.
As shown in figure 1, a monochromatic LED (1-3) of a third channel of the planar excitation light assembly emits light, stray light is filtered by an excitation light filter (2-3), the light is collimated by an excitation light collimating lens (3-3), then the light is reflected to the excitation light dichroic mirror (4-3) placed at 45 degrees from the excitation light dichroic mirror (4-2) placed at 45 degrees for transmission, the transmitted light reaches excitation light focusing lenses (5-1 and 5-2), then the light enters an incident optical fiber (6-1) through coupling, and the incident optical fiber (6-1) is connected to a reaction tank (7).
Light beams emitted by a sample in the reaction tank (7) are transmitted to the cylindrical lens (8) through the emergent optical fiber (6-2), the collimated light beams are reflected to the lighting dichroic mirror (9-2) arranged at 45 degrees from the lighting dichroic mirror (9-1) arranged at 45 degrees, the light beams pass through the lighting focusing lens (10-3) after being reflected, are focused and then penetrate through the lighting optical filter (11-3), and finally enter the surface of the third channel photoelectric sensor (12-3) of the lighting system.
As shown in figure 1, a monochromatic LED (1-4) of a fourth channel of the planar excitation light assembly emits light, stray light is filtered by an excitation light filter (2-4), the light is collimated by an excitation light collimating lens (3-4), then the light is transmitted to the excitation light dichroic mirror (4-3) placed at 45 degrees from the excitation light dichroic mirror (4-2) placed at 45 degrees, then the light is transmitted by the excitation light dichroic mirror (4-3) and then reaches excitation light focusing lenses (5-1 and 5-2), the light enters a light fiber (6-1) through light condensation coupling, and an incident optical fiber (6-1) is connected to a reaction tank (7).
The light beam excited by the sample in the reaction cell (7) is transmitted to the cylindrical lens (8) through the emergent optical fiber (6-2), the collimated light beam is reflected to the 45-degree lighting dichroic mirror (9-2) from the 45-degree lighting dichroic mirror (9-1), is transmitted through the lighting dichroic mirror (9-2), passes through the lighting focusing lens (10-4), is focused and then penetrates through the lighting optical filter (11-4), and finally enters the surface of the fourth channel photoelectric sensor (12-4).
Based on the scheme, the optical system comprises N channels, and light emitted by each LED light source passes through the same optical element. And the optical paths of the N channels are the same, so that the same attenuation of light of each channel can be ensured, the light intensity is convenient to control, and compared with other optical systems, the optical system does not need to additionally control the light intensity by a circuit system, and the function of the same light intensity can be directly achieved.
In addition, the light adopting the N-channel planar excitation light assembly and the planar lighting assembly only passes through the dichroic mirror for 2 times, so that the light beam passes through the dichroic mirror for the minimum time, and the control on the light beam propagation path is achieved.
Meanwhile, the N characteristics of a plurality of detected objects can be detected through the design of the N channels, and compared with a traditional optical system, the system enables the detection efficiency to be improved to the maximum.
The embodiments of the present invention are merely illustrative for explaining the principles and effects of the present invention, and are not intended to limit the present invention. Modifications and variations can be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be made by those skilled in the art without departing from the spirit and technical concepts of the present invention be covered by the claims of the present invention.
Claims (9)
1. The utility model provides a miniature multichannel fluorescence detection optical system of plane formula, is based on light source, lens subassembly, optic fibre, reaction tank and sensor, its characterized in that: including the following that do not interfere with each other: a planar excitation light assembly and a planar lighting assembly; the thin-wall tube is arranged in the reaction tank, and the reaction tank is connected to the planar excitation light assembly and the planar lighting assembly through optical fibers;
the planar excitation light assembly includes: the single LED light source light penetrates through the dichroic mirror as single-channel light after being filtered and collimated and/or is focused to the optical fiber after being reflected by the dichroic mirror;
single-channel light rays in different directions are output as a group of light ray bundles in the same emergent direction after passing through a first-stage dichroic mirror; the light ray bundles in different directions are output as a group of multi-channel light ray bundle groups in the same emergent direction after passing through a second-stage dichroic mirror;
the planar lighting assembly comprises: the fluorescence reaction penetrates through the dichroic mirror after passing through the cylindrical lens and/or is separated into a plurality of single-channel fluorescence in different directions after being reflected by the dichroic mirror, and the single-channel fluorescence is transmitted to a photosensitive surface of the photodiode after being focused and filtered;
the reaction fluorescence in the single direction is output as fluorescence beams in different emergent directions after passing through the dichroic mirror of the second stage; each group of fluorescent beams is output as single-channel fluorescent light in different emergent directions after passing through the first-stage dichroic mirror.
2. The planar micro multi-channel fluorescence detection optical system according to claim 1, wherein: the dichroic mirror comprises a transmission direction and a reflection direction; incident light rays in the penetrating direction and the reflecting direction form an incident angle of 45 degrees with the corresponding mirror surface.
3. The planar micro multi-channel fluorescence detection optical system according to claim 2, wherein: in the planar excitation light assembly: the two single-channel light rays form a group, and each group of single-channel light rays share a first-stage dichroic mirror; the two light beams form a group, and each group of light beams share one second-stage dichroic mirror.
4. The planar micro multi-channel fluorescence detection optical system according to claim 2, wherein: in the planar excitation light assembly: the dichroic mirror of the second stage separates two fluorescent beams in different emergent directions, and the dichroic mirror of the first stage separates single-channel fluorescent light in two different emergent directions.
5. The planar micro multi-channel fluorescence detection optical system according to claim 1, wherein: each of the planar excitation light assemblies includes four single-color LED lamps.
6. The planar micro multi-channel fluorescence detection optical system according to claim 1, wherein: comprises a plurality of reaction cells, a plurality of planar excitation light assemblies and a planar lighting assembly for detecting different characteristics of a plurality of objects to be detected.
7. The planar micro multi-channel fluorescence detection optical system according to claim 1, wherein: each light path in the planar excitation light assembly comprises the following components in sequence: monochromatic LED lamp, filter tube piece, collimating lens, dichroic mirror, focusing lens.
8. The planar micro multi-channel fluorescence detection optical system according to claim 1, wherein: every pipeline in the plane formula daylighting subassembly is gone up including passing through in proper order: cylindrical lens, dichroic mirror, focusing lens, optical filter.
9. The planar micro multi-channel fluorescence detection optical system according to claim 1, wherein: a standard 0.2ml thin-walled tube is placed in the reaction cell.
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| CN115629053A (en) * | 2022-08-24 | 2023-01-20 | 北京化工大学 | High-flux protein thermal stability analyzer |
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| CN115629053A (en) * | 2022-08-24 | 2023-01-20 | 北京化工大学 | High-flux protein thermal stability analyzer |
| CN115629053B (en) * | 2022-08-24 | 2024-05-24 | 北京化工大学 | High-flux protein thermal stability analyzer |
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Address after: 215000 rooms 101 and 201, C7 building, biomedical industrial park, 218 Xinghu street, Suzhou Industrial Park, Jiangsu Province Patentee after: Suzhou Yarui Biotechnology Co.,Ltd. Address before: 214500 1-2f, C7 building, 218 Xinghu street, Suzhou Industrial Park, Jiangsu Province Patentee before: SUZHOU MOLARRAY BIOTECHNOLOGY Co.,Ltd. |
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