CN102830224B - Optical path switching mechanism for homogeneous luminescent immunodetection - Google Patents
Optical path switching mechanism for homogeneous luminescent immunodetection Download PDFInfo
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- CN102830224B CN102830224B CN201210310778.3A CN201210310778A CN102830224B CN 102830224 B CN102830224 B CN 102830224B CN 201210310778 A CN201210310778 A CN 201210310778A CN 102830224 B CN102830224 B CN 102830224B
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- rotating disc
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- 230000003287 optical effect Effects 0.000 title abstract description 11
- 238000001514 detection method Methods 0.000 claims description 19
- 230000005284 excitation Effects 0.000 claims description 4
- 238000013461 design Methods 0.000 description 6
- 230000035945 sensitivity Effects 0.000 description 4
- 206010038743 Restlessness Diseases 0.000 description 3
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 3
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000012423 maintenance Methods 0.000 description 3
- 238000007634 remodeling Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
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- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
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- 230000001900 immune effect Effects 0.000 description 1
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- 238000002372 labelling Methods 0.000 description 1
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- 239000004005 microsphere Substances 0.000 description 1
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- 229910052760 oxygen Inorganic materials 0.000 description 1
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- 230000003071 parasitic effect Effects 0.000 description 1
- 239000003504 photosensitizing agent Substances 0.000 description 1
- 238000002310 reflectometry Methods 0.000 description 1
- 235000009165 saligot Nutrition 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention belongs to the technical field of homogeneous luminescent immunodetection and relates to an optical path switching mechanism for homogeneous luminescent immunodetection. The optical path switching mechanism for homogeneous luminescent immunodetection comprises a fixed holder (2) and a rotary disk (3). The fixed holder (2) is used for fixing a photo counter (1) and is provided with a signal light collection channel (11). A sample cell (7) is disposed below the rotary disk (3) and right below the signal light collection channel (11). The rotation center of the rotary disk (3) is eccentric to the center of the signal light collection channel. Rectangular prisms (6) for reflecting laser beams to irradiate the sample cell (7) and lenses (5) for collecting signal light are disposed at different angular positions on the rotary disk (3). Optical paths can be switched by controlling the rotary disk to rotate at different angles. The optical path switching mechanism is capable of lowering background noise in homogeneous luminescent immunodetection.
Description
Affiliated technical field
The invention belongs to homogeneous luminescent technical field of immunoassay, be specifically related to a kind of high speed light path switching mechanism.
Background technology
Homogeneous luminescent immunoassay technology is that of getting up of development in recent years is intended to replace existing enzyme mark technology, have efficient, high precision, high reliability, the brand-new clinical detection means of the series of advantages such as low cost.The utilization of homogeneous luminescent immunological technique is carried out the detection of biomolecule as the microballon of donor and acceptor.Wherein, donor microballon is filled with photosensitizer, and acceptor microballon is filled with chemiluminescence agent, if there is biomolecule to be measured in sample, donor microballon and acceptor microballon can be close to each other due to the specific binding of antibody antigen, thus form bridge couplings donor, be subject to attachment structure.After Stimulated Light excites, free oxygen transmits in bridge architecture, then produces the light signal of 615nm.Based on above principle, the core of homogeneous luminescent immune detection instrument is by photon counter, and semiconductor laser and high speed light path switching mechanism form.
Because sample produces flashlight after by laser excitation within very short time, and signal light intensity declines in time according to exponential law.Again due to the vulnerability of the high sensitivity of photon counter and target surface, during laser is to sample irradiation, any laser leaks into photon counter passage and all may cause detecting and unsuccessfully even damage expensive photon counter, has greatly reduced system signal noise ratio.Therefore, reasonably high speed light path switching mechanism is the key of system.
Existing light path switching mechanism is made up of optical shutter and sample carrier travel mechanism.Detecting point 4 steps completes: 1) sample cell is moved on to laser instrument below by travel mechanism, opens laser instrument sample cell is irradiated.During this period, optical shutter is closed, and stops photon to enter photon counter passage.2) Ear Mucosa Treated by He Ne Laser Irradiation finishes, and closes laser instrument.3) travel mechanism moves on to sample cell photon counter below at once, opens optical shutter and carries out photon counting.4) counted, closed optical shutter.
In the detecting step (1) of existing light path switching mechanism, be subject to the restriction of stepper motor-mechanical guide rail slide block structure its switching time, be difficult to the distance of tens millimeters of moving within a few tens of milliseconds, thereby miss optimum detection opportunity, reduce the sensitivity of system.Simultaneously, owing to being subject to the restriction of physical construction and material, the design of individual layer optical shutter is difficult to stop completely the scattered light of laser or external light source, and the scattered light of its infiltration will be leaked to photon counter passage, cause " dark counting " and ground unrest to raise, reduce the signal to noise ratio (S/N ratio) of system.
In the detecting step (3) of existing light path switching construction, another shortcoming of this mechanism is that optical shutter and travel mechanism are indispensable.In the occasion that does not need sample cell to move, as single-point device for fast detecting, the travel mechanism that only plays light path switching effect makes system complex, and cost increases.
In existing homogeneous luminescent immunity analysis instrument, LASER Light Source, light path switching mechanism, flashlight collection, photon counter all adopt Split type structure, be relatively independent between these four parts, cause whole instrument system complex structure, collaborative work Efficiency Decreasing, greatly increase instrument holistic cost, safeguarded and comparatively difficulty of upgrading.
Summary of the invention
The object of the invention is in order to reduce the ground unrest in homogeneous phase exciting light immune detection, avoid laser to leak into photon counter passage, develop a kind of high speed light path switching mechanism, and LASER Light Source, high speed light path switching mechanism, flashlight collection, photon counter are fixed to integrated design at the sub-position of interior key, making it to become a system core assembly independently, is the design of homogeneous phase exciting light immune detection system, produces, maintenance, remodeling brings great convenience.
As preferred implementation, for the light path switching mechanism of homogeneous luminescent immune detection, comprise fixed support 2 and rotating disc 3 for fixing photon counter 1, on fixed support, offer flashlight collection channel 11, sample cell 10 be placed in rotating disc 3 belows and in flashlight collection channel 11 under; Between the center of the rotation center of rotating disc 3 and flashlight collection channel, be eccentric position relation, be placed with for reflection lasering beam at the Difference angles place of rotating disc 3 and make laser beam irradiation to the right-angle prism 6 on sample cell 10 with for collecting the lens 5 of flashlight; Turn to respective angles by controlling rotating disc, make right-angle prism 6 and lens respectively the different periods rotate to flashlight collection channel 11 under, switch thereby realize light path.
As preferred implementation, the described light path switching mechanism for homogeneous luminescent immune detection, it is characterized in that, described fixed support 2 and the combination end face of rotating disc 3 are provided with concentric annular optics labyrinth structure, in this structure, below fixed support 2 He above rotating disc 3, be provided with multinest formula protuberance and groove structure, protuberance and groove intercouple, but do not hinder the rotation of rotating disc; Described rotating disc 3 in the time of laser excitation station, rotates to right-angle prism 6 above sample cell 10 under stepper motor drives, and the light beam 8 that laser instrument 7 sends is done after 90 degree turn to be radiated at sample cell 10 through right-angle prism 6; Described rotating disc 3 in the time of input station, rotates to flashlight collecting lens 5 above sample cell 10 under stepper motor drives; Photon counter 1, the stationary shaft of laser instrument 7 and rotating disc 3 is all integrated in fixed support 2) on.
The invention has the beneficial effects as follows by the rotation of rotating disc and realize light path from exciting station to the switching that detects station, and by concentric annular labyrinth structure being set on the combination end face between rotating disc and fixed support, effectively reduce the ground unrest in homogeneous phase exciting light immune detection, avoided laser to leak into photon counter passage.In addition the present invention is needed homogeneous luminescent immune detection photon counter, laser instrument, and rotating disc, flashlight sampler is all integrated on fixed support at interior crucial subassembly, makes it to become a system core assembly independently.This individual cores assembly is the design of homogeneous luminescent immune detection system, produces, and maintenance, remodeling brings great convenience.
Brief description of the drawings
The structural drawing of Fig. 1 high speed light path switching mechanism, (a) and (b) is respectively rotating disc position view while exciting station and while detecting station; (c) and (d) be respectively the light channel structure schematic diagram while exciting station and while detecting station.
Fig. 2 is the optics labyrinth multiple reflections schematic diagram that the present invention adopts.
Number in the figure explanation:
1: photomultiplier/photon counter
2: fixed support
3: rotating disc
4: lens 2
5: lens 1
6: right angle water chestnut mirror
7: laser instrument
8: laser beam
9: optics labyrinth ring
10: sample cell
11: flashlight collection channel
Embodiment
The present invention is a kind of high speed light path switching mechanism, comprises photon counter (1), fixed support (2), rotating disc (3) and laser instrument (7) etc.Photon counter (1) is arranged on fixed support (2).On fixed support (2), have the flashlight collection channel (11) concentric with photon counter target surface.Sample cell (10) is placed in the below of rotating disc (3) the position concentric with flashlight collection channel (11).The rotation center of rotating disc (3) and flashlight collection channel (11) center are set for the eccentric.Right-angle prism (6) for reflection lasering beam is installed respectively on the diameter that rotating disc (3) overlaps with flashlight collection channel (11) and for collecting the lens (5) of flashlight, sees accompanying drawing 1.
Comprise photon counter (1), laser instrument (7), rotating disc (3), it is upper that flashlight sampler is all integrated in fixed support (2) at interior crucial subassembly, makes it to become a high speed light path switching mechanism independently.This individual cores assembly is the design of homogeneous luminescent immune detection system, produces, and maintenance, remodeling brings great convenience.
The combination end face of fixed support (2) and rotating disc (3) is concentric annular optics labyrinth structure.In this structure, be provided with multinest formula protuberance and groove structure in fixed support (2) below and rotating disc (3) top, the mutual close-coupled of protuberance and groove also can freely be rotated, thereby forms multilayer barrier labyrinth structure.See accompanying drawing 2, Δ is enough little, and scattered light need to experience and reflect many times the surface that could arrive photon counter.The surface reflectivity in optics labyrinth is enough low, and the scattered light intensity of multiple reflections declines with the form of index, and the photon number that arrives photon counter target surface can be lower than the background noise sound of photon counter itself.
Referring to accompanying drawing 1, the principle of work of high speed light path switching mechanism is: (1) is at laser excitation station, rotating disc (3) rotates to right-angle prism (6) sample cell (10) top under stepper motor drives, and the light beam (8) that laser instrument (7) sends is done after 90 degree turn to be radiated at sample cell through right-angle prism (6).Because right-angle prism (6) does not communicate with flashlight collection channel (11) at the fixing position of rotating disc (3), flashlight collecting lens (5) is also rotated to the angle not communicating with flashlight collection channel (11), add the catadioptric absorption function in concentric annular optics labyrinth, the scattered light that irradiates sample can not drain to the collection channel of photon counter.Low-down system " dark counting " value and background noisiness are ensured.(2), at input station, rotating disc (3) rotates to flashlight collecting lens (5) sample cell (10) top under stepper motor drives.Now, sample cell (10), flashlight collecting lens (5), flashlight collection channel (11), flashlight collecting lens (4) and photon counter target surface are concentric, and photon counting passage is opened.
Because the moment of inertia of rotating disc (3) can design much smaller than the translation inertia of sample cell movable support bracket, light path is switched and can within a few tens of milliseconds, be completed.
Further set forth integrated high speed light path switching mechanism below in conjunction with experimental data and ensureing, on the basis of low noise, low stray light, can also to obtain higher detection sensitivity.
Experiment condition
At standard 96 orifice plate reacting holes, (lucifuge deep hole approximately 300 μ mix the coated acceptor microspheres solution (10 μ g/ml) of 25 μ l biotin labeling antibody+25, μ l alpha-fetoprotein (AFP) standard model+25 μ l antibody in l), after above-mentioned solution is mixed, at 37 DEG C of concussion incubation 30min.Lucifuge adds the 175 general liquid of μ l donor microballoon (self-control), fully mixes concussion incubation 10min.
Experimental result
In experiment, the concentration of AFP standard model is designed to 0,10,20,50,100,250ng/ml.Adopt the corresponding data of high speed light path switching mechanism collection, as shown in the table.From table, adopt high speed light path switching mechanism can obviously end parasitic light, reduce nature background fluorescence, significantly improve the sensitivity of system.
Sample concentration | 0 | 10 | 20 | 50 | 100 | 250 |
Signal intensity | 28 | 1806 | 3480 | 6705 | 9094 | 9275 |
Claims (5)
1. the light path switching mechanism for homogeneous luminescent immune detection, comprise fixed support (2) and rotating disc (3) for fixing photon counter (1), on fixed support, offer flashlight collection channel (11), sample cell (10) be placed in rotating disc (3) below and in flashlight collection channel (11) under; Between the rotation center of rotating disc (3) and the center of flashlight collection channel, be eccentric position relation, be placed with for reflection lasering beam at the Difference angles place of rotating disc (3) and make laser beam irradiation to the right-angle prism (6) on sample cell (10) with for collecting the lens (5) of flashlight; Turn to respective angles by controlling rotating disc, make right-angle prism (6) and lens respectively the different periods rotate to flashlight collection channel (11) under, thereby realize light path switching.
2. the light path switching mechanism for homogeneous luminescent immune detection according to claim 1, it is characterized in that, described fixed support (2) and the combination end face of rotating disc (3) are provided with concentric annular optics labyrinth structure, in this structure, be provided with multinest formula protuberance and groove structure in fixed support (2) below and rotating disc (3) top, protuberance and groove intercouple, but do not hinder the rotation of rotating disc.
3. the light path switching mechanism for homogeneous luminescent immune detection according to claim 1, it is characterized in that, described rotating disc (3), in the time of laser excitation station, right-angle prism (6) is rotated to sample cell (10) top under stepper motor drives, the light beam (8) that laser instrument (7) sends is done after 90 degree turn to be radiated at sample cell (10) through right-angle prism (6).
4. the light path switching mechanism for homogeneous luminescent immune detection according to claim 1, it is characterized in that, described rotating disc (3), in the time of input station, flashlight collecting lens (5) is rotated to sample cell (10) top under stepper motor drives.
5. the light path switching mechanism for homogeneous luminescent immune detection according to claim 1, it is characterized in that, photon counter (1), the stationary shaft of laser instrument (7) and rotating disc (3) is all integrated on fixed support (2).
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CN110146493B (en) * | 2018-02-11 | 2024-09-27 | 科美诊断技术(苏州)有限公司 | Light path detection system and light excitation chemiluminescence detector with same |
CN109669038B (en) * | 2019-02-28 | 2024-06-21 | 深圳市易瑞生物技术股份有限公司 | Portable multichannel chromatographic device |
CN111443042B (en) * | 2020-06-01 | 2025-01-10 | 厦门爱科思光科技有限公司 | A device for measuring variable temperature long afterglow characteristics |
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JPH09237428A (en) * | 1995-12-27 | 1997-09-09 | Sony Corp | Biaxial actuator and optical disk device |
DE19815109A1 (en) * | 1998-04-03 | 1999-10-07 | Bodenseewerk Perkin Elmer Co | Device for detecting a fluorescent dye |
CN1120361C (en) * | 1999-07-08 | 2003-09-03 | 上海春晓光电科技有限公司 | Optical system of multifunction spectrophotometer |
JP2001091463A (en) * | 1999-09-22 | 2001-04-06 | Tosoh Corp | Scanner-type fluorescence detector using a small excitation light source |
DE10136863A1 (en) * | 2001-07-28 | 2003-02-20 | Berthold Tech Gmbh & Co Kg | Device for the optional measurement of in particular luminescence and / or fluorescence radiation |
JP2003262804A (en) * | 2002-03-11 | 2003-09-19 | Teijin Seiki Co Ltd | Optical path switching device |
JP2005062023A (en) * | 2003-08-14 | 2005-03-10 | Japan Science & Technology Agency | Epi-illumination fluorescence measuring device |
US7295316B2 (en) * | 2004-01-14 | 2007-11-13 | Applera Corporation | Fluorescent detector with automatic changing filters |
JP2006010515A (en) * | 2004-06-25 | 2006-01-12 | Aisin Seiki Co Ltd | Fluorescence intensity detector |
JP3949681B2 (en) * | 2004-09-03 | 2007-07-25 | 株式会社トプコン | Rotating laser device |
CN100427923C (en) * | 2006-03-08 | 2008-10-22 | 东华大学 | Multifunctional chip detection device |
JP4644261B2 (en) * | 2007-09-10 | 2011-03-02 | 中部日本マルコ株式会社 | Non-contact connector |
CN102435314B (en) * | 2011-11-22 | 2013-09-11 | 中国科学院安徽光学精密机械研究所 | Multi-optical-path switching system for satellite-borne differential absorption measuring instrument |
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