CN1766580A - Scanning type up-conversion phosphor particle distribution state detector - Google Patents
Scanning type up-conversion phosphor particle distribution state detector Download PDFInfo
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- CN1766580A CN1766580A CN 200510030550 CN200510030550A CN1766580A CN 1766580 A CN1766580 A CN 1766580A CN 200510030550 CN200510030550 CN 200510030550 CN 200510030550 A CN200510030550 A CN 200510030550A CN 1766580 A CN1766580 A CN 1766580A
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Abstract
A scanning type up-conversion phosphor particle distribution state detector comprises: the device comprises a photoelectric conversion and signal processing system, a data acquisition and control system, a phosphorescence excitation light path and a phosphorescence receiving light path, wherein the phosphorescence excitation light path comprises an infrared semiconductor laser, and a spherical surface adjusting mirror, a dichroic mirror, a focusing/collimating mirror and a scanning platform are sequentially arranged along the advancing direction of an excitation light beam emitted by the infrared semiconductor laser, and the scanning platform is used for placing a test strip which is positioned on the focal plane of the focusing/collimating mirror; the phosphorescence receiving light path consists of the focusing/collimating mirror and the dichroic mirror which share the optical axis, and a filter, a rear focusing mirror, a confocal slit diaphragm and a photoelectric converter which are arranged in sequence; the confocal slit diaphragm is positioned on the back focal plane of the back focusing mirror. The invention has the advantages of high detection sensitivity, high detection reliability, high detection spatial resolution, high detection stability and sterilization function, thereby ensuring the safety of detection personnel.
Description
Technical field
The present invention relates to immunochromatography technique, particularly a kind of scanning type up-converting phosphor particle distribution state detector.This detector can detect and interpretation as a result scanning based on last converting phosphor technology immune chromatography test paper, thereby realizes quick, multiple, detection by quantitative to plurality of target checking matters such as pathogen, antigen, antibody, illegal drug, major disease (tumour, cancer and diabetes etc.) marks.
Background technology
At present, in the immunochromatography technique, usually with enzyme, collaurum and the things that serve as a mark such as coloring agent pearl, with test strips as detecting carrier, usually adopt the detection band of test strip such as visual method, CCD and cmos image sensor and the luminous intensity of the label on the quality control band, thereby determine the concentration of target checking matter in the sample.
There is following shortcoming in above-mentioned technology formerly:
1. detection sensitivity is low.The general light emitting diode that adopts is as lighting source, and illumination intensity is low; When receiving image, need large area lighting, thereby tested paper slip surface is difficult to obtain high intensity illumination with face formation CCD or cmos image sensor; And the sensitivity of face formation CCD or cmos image sensor is lower usually.If observe with visual method, then except that sensitivity was low, testing result also was subjected to artificial factor.
2. the reliability of testing result is low.Because the label luminous intensity on detected test strips surface is directly proportional with illumination light intensity, in order to obtain stable testing result, these Technology Need large tracts of land are evenly thrown light on, and the even illumination of large tracts of land is difficult to realize usually.
3. because the restriction of detection sensitivity generally can only be carried out qualitative analysis, can't realize that accurate quantification detects;
4. some label as glue Jin Ti etc., has certain life-span.Thereby, can't preserve the long period by the test strips that this label is made, so can not check;
5. do not have sterilizing function, when testing result is positive, may work the mischief the testing staff.
Summary of the invention
The objective of the invention is to overcome the shortcoming of above-mentioned prior art, a kind of scanning type up-converting phosphor particle distribution state detector is provided, this detector should have detection sensitivity height, detecting reliability height, detects the spatial resolution height, detects the stability height and have the function of sterilizing, to ensure testing staff's safety.
Technical solution of the present invention is as follows:
A kind of scanning type up-converting phosphor particle distribution state detector, comprise optical system, scanning platform, opto-electronic conversion and signal processing system, Data Acquisition and Conversion System (DACS), described opto-electronic conversion and signal processing system are made of photoelectric commutator and prime amplifier, described Data Acquisition and Conversion System (DACS) is made of multifunctional data acquisition card, embedded computer and process software, it is characterized in that:
Described optical system is made up of phosphorescent illumination and phosphorescence receiving light path, described phosphorescent illumination comprises an infrared semiconductor laser, on the excitation beam working direction of sending along this infrared semiconductor laser, be provided with sphere successively and adjust mirror, dichronic mirror, focusing/collimating mirror and scanning platform, this scanning platform is for placing test strips, and this test strips is positioned on the focal plane of focusing/collimating mirror; Described phosphorescence receiving light path is made up of the described focusing/collimating mirror of common optical axis and dichronic mirror and the optical filter that sets gradually, back focus lamp, confocal slit aperture, photoelectric commutator; Described confocal slit aperture is positioned on the back focal plane of back focus lamp; Described phosphorescent illumination part before described dichronic mirror is vertical mutually with the optical axis of phosphorescence receiving light path, the surface of this dichronic mirror and the optical axis of described phosphorescent illumination angle at 45, phosphorescent illumination behind this dichronic mirror with described phosphorescence receiving light path common optical axis.
Be provided with ultraviolet germicidal between described test strips and the described focusing/collimating mirror, and place on described phosphorescent illumination and the unobstructed position of phosphorescence receiving light path, can be at any time to tested paper slip carry out disinfection, sterilization processing.
Described dichronic mirror is coated with to the reflectance coating of exciting light with to the anti-reflection film of phosphorescence.
Described sphere is adjusted mirror and is provided with along the adjustment structure of the position of optical axis direction.
Described photoelectric commutator is a photomultiplier.
(Up-converting phosphor, UCP) immuno-chromatographic test paper strip (abbreviation test strips) as biomarker is a detected object of the present invention to converting phosphor material in the employing.On test strips, be provided with a plurality of function bands, as a quality control band and one or more detection band.The function band is arranged in parallel with a determining deviation, and its direction is perpendicular to the test strips long side direction.The present invention with the infrared semiconductor laser of firm power as excitation source, scan detection to detecting on band and the quality control band on the test strips by the UCP particle stimulated luminescence intensity on the biological immune reaction bonded, determine its content and distribution, according to the corresponding relation of photosignal and biological information, realize quick, multiple, quantitative test to one or more target checking matters.
Described Data Acquisition and Conversion System (DACS) is made of the multifunctional data acquisition card that possesses high-precision A/D translation function, embedded computer and process software.In process software, adopt suitable algorithm that data are handled and analyzed, provide sentence read result.
Compare with technology formerly, the present invention has following technique effect:
1. detection sensitivity height.The present invention adopts infrared semiconductor laser as excitation source, and is focused into focal line, the excitating light strength height; The phosphorescence receiving light path adopts the optical texture of confocal pattern, has reduced the influence of background noise.
2. detecting reliability height.It is constant that the power of excitation source of the present invention keeps, and need not the even lighting requirement in the large tracts of land.
3. detect the spatial resolution height.The present invention scans detection with superfine focal line to the UCP distribution of particles on the tested paper slip.
4. it is high to detect stability.Because the luminescent properties of UCP is invariable, so but with UCP particle serve as a mark after the test strips long preservation that thing makes still duplicate detection and testing result stable and consistent.
5. ultraviolet germicidal is housed, occurs using under the positive situation, can ensure testing staff's safety in testing result.
Description of drawings
Fig. 1 is a structural representation of going up the converting phosphor immuno-chromatographic test paper strip.
Fig. 2 is the structure and the light path synoptic diagram of scanning type up-converting phosphor particle distribution state detector most preferred embodiment of the present invention.
Fig. 3 is the stimulated emission curve of spectrum of the UCP particle that adopts of the present invention.
Fig. 4 is the curve of spectrum of the infrared semiconductor laser that adopts of the present invention.
Fig. 5 is the transmittance curve of the dichronic mirror that adopts of the present invention.
Fig. 6 is the transmittance curve of the optical filter that adopts of the present invention.
Fig. 7 is that the present invention is to standard series concentration test result of samples.
Fig. 8 is the examination testing result that the present invention carries out 239 parts of blood serum samples.
Embodiment
See also Fig. 2, Fig. 2 is the structure and the light path synoptic diagram of scanning type up-converting phosphor particle distribution state detector most preferred embodiment of the present invention.As seen from the figure, scanning type up-converting phosphor particle distribution state detector of the present invention is made up of optical system, scanning platform 2, opto-electronic conversion and signal processing system, Data Acquisition and Conversion System (DACS); Described opto-electronic conversion and signal processing system are made of photoelectric commutator 10 and prime amplifier 11, described Data Acquisition and Conversion System (DACS) is by multifunctional data acquisition card 12, embedded computer 13 and process software constitute, it is characterized in that: described optical system is made up of phosphorescent illumination and phosphorescence receiving light path, described phosphorescent illumination comprises an infrared semiconductor laser 3, on the excitation beam working direction of sending along this infrared semiconductor laser 3, be provided with sphere successively and adjust mirror 4, dichronic mirror 6, focusing/collimating mirror 5 and scanning platform 2, this scanning platform 2 is for placing test strips 1, and this test strips 1 is positioned on the focal plane of focusing/collimating mirror 5; Described phosphorescence receiving light path is made up of the described focusing/collimating mirror 5 of common optical axis and dichronic mirror 6 and the optical filter 7 that sets gradually, back focus lamp 8, confocal slit aperture 9, photoelectric commutator 10; Described confocal slit aperture 9 is positioned on the back focal plane of back focus lamp 8; Described phosphorescent illumination part before described dichronic mirror 6 is vertical mutually with the optical axis of phosphorescence receiving light path, the surface of this dichronic mirror 6 and the optical axis of described phosphorescent illumination angle at 45, phosphorescent illumination behind this dichronic mirror 6 with described phosphorescence receiving light path common optical axis.Between described test strips 1 and described focusing/collimating mirror 5, be provided with ultraviolet germicidal 14, and be on described phosphorescent illumination and the unobstructed position of phosphorescence receiving light path.Described dichronic mirror 6 is coated with to the reflectance coating of exciting light with to the anti-reflection film of phosphorescence.Described sphere is adjusted mirror (4) and is provided with along the adjustment structure of the position of optical axis direction.Described photoelectric commutator 10 is a photomultiplier.
Detected object is that immunoreactive test strips 1 has taken place, and the structure of test strips 1 as shown in Figure 1.The present invention by test strips 1 is detected with 101 and quality control band 102 on UCP particle stimulated luminescence intensity scan detection, analyze its content and distribution, thereby derive the concentration information of target checking matter in the test sample.
Optical system of the present invention comprises phosphorescent illumination and phosphorescence receiving light path, is respectively applied for the UCP particle that excites on the test strips 1, and receives the phosphorescent signal that the UCP particle sends; Scanning platform 2 is used to carry test strips 1, finishes the shuttle-scanning motion under the control of described embedded computer 13; Opto-electronic conversion and signal processing system are used for the phosphorescent signal switching electrical signals that will receive, and are amplified to certain amplitude; The scanning that Data Acquisition and Conversion System (DACS) is used to control test strips 1 detects motion, and electric signal is gathered, analyzed and handles, and obtains net result.
The infrared beam that infrared semiconductor laser 3 sends is focused into a focal line on test strips 1 surface after sphere is adjusted mirror 4, dichronic mirror 6 and focusing/collimating mirror 5.Sphere is adjusted mirror 4 can carry out trace adjustment along optical axis direction, with the length and the width of the focal line that obtains suitable illumination.The length of focal line and width should be complementary with test strips width and scanning resolution of the present invention respectively, have advantages of higher stability and sensitivity to guarantee detector.
The optical axis of phosphorescent illumination is vertical mutually with the optical axis of receiving light path, and all with the angle surperficial at 45 of dichronic mirror 6, make two light path common optical axis by dichronic mirror 6.This design has reduced the dimensions of mechanical structures of light path.
Being excited of UCP particle in the focal line scope of exciting light illumination and converting phosphor on producing, line focus/collimating mirror 5 collimations successively behind dichronic mirror 6 and optical filter 7 filtering exciting lights and other parasitic lights, are focused on the confocal slit aperture 9 by back focus lamp 8 again.The suitable dimensions of choosing this confocal slit aperture 9 can reduce outside noise to be disturbed, and improves detection sensitivity.
Phosphorescent signal by confocal slit aperture 9 is converted to electric signal by electrooptical device 10, amplify through prime amplifier 11, convert digital signal to by the multifunctional data acquisition card 12 that possesses the A/D translation function again, gather, carry out data processing, storage by embedded computer 13.
Embedded computer 13 is by the round scanning motion of multifunctional data acquisition card 12 output pulse signals and direction signal gated sweep platform 2, thereby realization is to the scanning detection of UCP phosphorescent signal distribution on the test strips 1.
Process software adopts following algorithm that the signal that collects is handled: respectively get 10 to 20 points in detection with before and after 101 the central point, promptly get 21 to 41 signal value altogether, calculate the mean value of these points, with this mean value as detection with 101 signal value T; Before and after the central point of quality control band 102, respectively get 10 to 20 points, promptly get 21 to 41 signal value altogether, calculate the mean value of these points, with the signal value C of this mean value as quality control band 102; Calculate to detect the ratio T/C of band signal value T and quality control band signal value C, T/C is corresponding with the concentration of target checking matter, compares by standard working curve, calculates the concentration of target checking matter.
Compare with technology formerly, characteristics of the present invention are:
1. excitation source adopts the infrared semiconductor laser 3 of power stabilize;
2. utilize sphere adjustment mirror 4 to move, the exciting light focal line size on control test strips 1 surface along the trace of optical axis direction;
3. adopt dichronic mirror 6 to make phosphorescent illumination and phosphorescence receiving light path common optical axis, reduced the size of optical system;
4. the phosphorescence receiving light path adopts the optical texture of confocal pattern, has reduced the influence of background noise, has improved detection sensitivity;
5. utilize scan method to detect, the spatial resolution height, low to the uniformity requirement of excitation source, can realize multiple, detection by quantitative.
Compare with other light sources, said excitation source adopts infrared semiconductor laser 3, has that volume is little, power is high, the life-span is long, good directionality and good characteristics of monochromaticity.Do not occurring absorbing under the saturated conditions, the stimulated luminescence intensity of UCP particle is in rising trend with the increase of excitating light strength in the certain limit.The wavelength of infrared semiconductor laser 3 is identical or close with selected UCP particle absorption peak wavelength, to improve the efficient of UCP particle stimulated luminescence.
Through behind the phosphorescent illumination, excitation beam is focused into focal line.This focal line size can be adjusted mirror 4 by sphere and adjust.Its width dimensions direction is consistent with the direction of scanning of scanning platform 2, and focal line width size equals or be slightly larger than the step distance of scanning platform 2.If this is undersized, the distributed intelligence meeting of UCP particle is lost; If this is oversize, the intensity of exciting light focal line can reduce.The width of focal line length dimension and test strips 1 is close, to avoid UCP distribution of particles information dropout.
The effect of dichronic mirror 6 is reflection exciting lights, sees through phosphorescence, therefore to exciting light high as far as possible reflectivity should be arranged; The effect of optical filter 7 is filtering parasitic lights, and the principal ingredient of parasitic light is by the micro-exciting light of test strips 1 surface reflection through dichronic mirror 6 transmissions, so 7 pairs of exciting lights of optical filter should have the high as far as possible efficient of ending.And dichronic mirror 6 all should have high as far as possible transmitance with 7 pairs of phosphorescence of optical filter.
The confocal slit aperture 9 that the present invention adopts only makes the test strips 1 surface phosphorescence that the UCP particle produces in the zone that is stimulated pass through, phosphorescence or parasitic light that test strips 1 surperficial upper and lower dust granule produces are focused on outside the confocal slit aperture 9, further the interference of the extraneous parasitic light of filtering has improved detection sensitivity.
The course of work of scanning type up-converting phosphor particle distribution state detector of the present invention is as follows:
Test strips to be detected 1 is positioned on the scanning platform 2, and the laser beam of being sent by infrared semiconductor laser 3 is reflected by dichronic mirror 6 after sphere is adjusted mirror 4, and line focus/collimating mirror 5 backs form focal line on test strips 1 surface again.The phosphorescence that UCP particle in the focal line scope sends is focused/collimating mirror 5 receives and collimation, behind dichronic mirror 6 and optical filter 7 filtering parasitic lights, again through after focus lamp 8 focus on the confocal slit aperture 9.Phosphorescent signal by confocal slit aperture 9 is converted to electric signal by electrooptical device 10, amplifies through prime amplifier 11, converts digital signal to by the multifunctional data acquisition card 12 that possesses the A/D translation function again, enters embedded computer 13 and gathers, stores.Embedded computer 13 is by the round step motion of multifunctional data acquisition card 12 output pulse signals and direction signal gated sweep platform 2, test strips 1 result scanned the UCP phosphorescent signal scans detection in the window, embedded computer 13 goes out to detect with 101 signal value T, the signal value C and the ratio between two T/C of quality control band 102 by certain algorithm computation, calculates the concentration of target checking matter at last by standard working curve.
Fig. 2 is the structural representation of most preferred embodiment of the present invention, enumerates its concrete structure and parameter below:
The UCP material that the present invention adopts is NaYF
4: Yb
3+, Er
3+, Fig. 3 is excitation wavelength its spectral radiation curves when being 980nm, and emission light main peak value wavelength is 541.5nm, and the secondary peak wavelength is 669.6nm.
The centre wavelength of the infrared semiconductor laser 3 that the present invention selects for use is 980nm, and power is 30mW, its curve of spectrum as shown in Figure 4, its peak wavelength is at the 980nm place.
The clear aperture that sphere in the phosphorescent illumination is adjusted mirror 4 is φ 8mm, and focal length is 20mm.
Focusing/collimating mirror 5 is selected planoconvex lens for use, and its clear aperture is φ 14.6mm, and focal length is 20.64mm.Infrared excitation light focuses on test strips 1 surface behind phosphorescent illumination focal line is of a size of 2mm * 27 μ m.This size is complementary with the width 3mm of test strips 1 and the step resolution 20 μ m of scanning platform 2 respectively.
The reflecting surface of dichronic mirror 6 plates the film that is all-trans to the 980nm wavelength, plates anti-reflection film in the stimulated emission phosphorescence wavelength coverage of transmission plane to the UCP material.Fig. 5 and 6 is respectively the transmittance curve of dichronic mirror 6 and optical filter 7.Dichronic mirror 6 is about 0.88% in the transmitance at 980nm place, and the transmitance in the phosphorescence wavelength coverage is between 85～95%; Optical filter 7 greater than 85%, and approaches 0 in the transmitance at 980nm place in 541.5nm place transmitance.
Embedded computer 13 is the PC104 embedded system, and multifunctional data acquisition card 12 has A/D, D/A and DI/DO function, and signals collecting/output frequency is 100kHz.
The power of ultraviolet germicidal 14 is 5W, can kill various viruses and bacterium fully in 20min.
The mode of present embodiment being carried out study on the stability is: to same test strips 1 to be checked after the start, after the long-term work and repeat to plug under three kinds of situations of test strips and carry out repeatedly duplicate detection.Adopt T/C as testing result judge value, its Variation Lines number average illustrates working stability of the present invention less than 3～5% in all cases.
The last converting phosphor immunity test strip that present embodiment obtains after to the reaction of 20 parts of series concentration standard items detects, scanning detecting result with the T/C value as X-axis, with the normal concentration of the anti-plague IgG of rabbit as Y-axis, drawing standard concentration sample detection figure, as shown in Figure 7.As shown in Figure 7, in the 200-6000ng/ml concentration range, the linear response relationship of present embodiment is better.Can find out also that simultaneously the detection sensitivity of detector has been better than minimum positive criteria sample concentration (200ng/ml).
Adopt least square method that the data in the 200-6000ng/ml concentration range are carried out linear fit, the straight line that obtains has very high correlativity, its coefficient R
2〉=0.95.
The test strips that present embodiment obtains after to the reaction of 239 parts of human serums has been carried out the detection of anti-plague antibodies, the results are shown in Figure 8, at this with the net result of T/C value as detection by quantitative.This result shows that present embodiment has possessed the ability of Yersinia pestis antibody being carried out actual detected, and highly sensitive.
Present embodiment has been finished detection by quantitative to anthrax spore virus in 30 minutes, multiple sporeformer and other pathogen are estimated the detection sensitivity that shows present embodiment and reach 1 * 10
4Individual gemma.
Claims (5)
1, a kind of scanning type up-converting phosphor particle distribution state detector is made up of optical system, scanning platform (2), opto-electronic conversion and signal processing system, Data Acquisition and Conversion System (DACS); Described opto-electronic conversion and signal processing system are made of photoelectric commutator (10) and prime amplifier (11), described Data Acquisition and Conversion System (DACS) by multifunctional data acquisition card (12), embedded computer (13 and process software constitute, it is characterized in that:
Described optical system is made up of phosphorescent illumination and phosphorescence receiving light path, described phosphorescent illumination comprises an infrared semiconductor laser (3), on the excitation beam working direction of sending along this infrared semiconductor laser (3), be provided with sphere successively and adjust mirror (4), dichronic mirror (6), focusing/collimating mirror (5) and scanning platform (2), this scanning platform (2) is for placing test strips (1), and this test strips (1) is positioned on the focal plane of focusing/collimating mirror (5); Described phosphorescence receiving light path is made up of the described focusing/collimating mirror (5) of common optical axis and dichronic mirror (6) and the optical filter (7) that sets gradually, back focus lamp (8), confocal slit aperture (9), photoelectric commutator (10); Described confocal slit aperture (9) is positioned on the back focal plane of back focus lamp (8); Part is vertical mutually with the optical axis of phosphorescence receiving light path before at described dichronic mirror (6) for described phosphorescent illumination, the optical axis angle at 45 of surface of this dichronic mirror (6) and described phosphorescent illumination, phosphorescent illumination behind this dichronic mirror (6) with described phosphorescence receiving light path common optical axis.
2, scanning type up-converting phosphor particle distribution state detector according to claim 1, it is characterized in that being provided with ultraviolet germicidal (14) between described test strips (1) and the described focusing/collimating mirror (5), and be positioned on described phosphorescent illumination and the unobstructed position of phosphorescence receiving light path.
3, scanning type up-converting phosphor particle distribution state detector according to claim 1 is characterized in that described dichronic mirror (6) is coated with to the reflectance coating of exciting light with to the anti-reflection film of phosphorescence.
4, scanning type up-converting phosphor particle distribution state detector according to claim 1 is characterized in that described sphere adjustment mirror (4) is provided with along the adjustment structure of the position of optical axis direction.
5, scanning type up-converting phosphor particle distribution state detector according to claim 1 is characterized in that described photoelectric commutator (10) is a photomultiplier.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100424492C (en) * | 2006-05-26 | 2008-10-08 | 中国科学院上海技术物理研究所 | Narrowband two-waveband scanning-type infrared focal plane detector |
| CN103149353A (en) * | 2013-02-05 | 2013-06-12 | 中国科学院上海光学精密机械研究所 | Quantitative detection system and detection method for multi-channel test paper disc |
| CN104508483A (en) * | 2012-03-01 | 2015-04-08 | 奎多公司 | Interactive test device and apparatus with timing mechanism |
| CN105353115A (en) * | 2015-10-21 | 2016-02-24 | 中国科学院上海光学精密机械研究所 | Measuring device and method of spatial distribution of scattering light field of immunochromatographic test strip |
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| CN109791040A (en) * | 2016-08-25 | 2019-05-21 | 纳诺福卡斯股份公司 | Method and apparatus for carrying out optical surface measurement by colored confocal sensor |
| CN110568179A (en) * | 2019-11-11 | 2019-12-13 | 上海奥普生物医药有限公司 | device and method for realizing multi-line extensible detection of immunochromatography analyzer |
| US10845300B2 (en) | 2013-12-06 | 2020-11-24 | Quidel Corporation | Method for reducing analyzer variability using a normalization target |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100424492C (en) * | 2006-05-26 | 2008-10-08 | 中国科学院上海技术物理研究所 | Narrowband two-waveband scanning-type infrared focal plane detector |
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