CN201262615Y - Rapid detection device for pathogenic microorganism - Google Patents
Rapid detection device for pathogenic microorganism Download PDFInfo
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- CN201262615Y CN201262615Y CN 200820099973 CN200820099973U CN201262615Y CN 201262615 Y CN201262615 Y CN 201262615Y CN 200820099973 CN200820099973 CN 200820099973 CN 200820099973 U CN200820099973 U CN 200820099973U CN 201262615 Y CN201262615 Y CN 201262615Y
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Abstract
The utility model discloses a rapid detection device of pathogenic microorganism, which comprises an SPR biosensor detection device, an incident light channel, an emergent light channel and an intelligent analysis processing platform; the SPR biosensor detection device comprises an SPR biosensor and an adjustable detection light source; the incident light channel comprises a cruise adjustable polarized light source which includes a polarized light source and a control system thereof; the emergent light channel comprises a convergent lens and a photodetector; the rapid detection device can accurately analyze the mutual action (multi-factor action) among various substances in a sample, avoids the system deviation caused by single measurement, greatly reduces the detection time and accelerates the detection process; the rapid detection device is easy for carry and high in detection sensitivity, can be applied to a plurality of fields and various environments, particularly can fully meet the need of use in the wild environment and has broad application prospect.
Description
Technical field
The utility model relates to field of biosensors, particularly a kind of pathogenic microorganism device for fast detecting.
Background technology
Surface plasma body resonant vibration (SPR) is a kind of physical optics phenomenon.It is a kind of technology of utilizing surface plasma wave (SPW, Surface Plasma Wave) to detect that surface plasma body resonant vibration detects, advantages such as having need not mark, high speed, selectivity, highly sensitive and massive parallel screening.Surface plasma (SP) is that the electromagnetic wave of propagating along interface between metal and dielectric forms.When the polarized light of parallel surfaces is incident on the interface to be referred to as the surface plasma body resonant vibration angle, when attenuated total reflection takes place, incident light is coupled in the surface plasma, and luminous energy is absorbed in a large number, causes that owing to surface plasma body resonant vibration boundary reflection light significantly reduces on this angle.Because SPR is very responsive to the dielectric refractive index in metal surface, its surface plasma body resonant vibration angle difference of different dielectrics.Dielectric of the same race, it is attached to the amount difference of metal surface, then the response intensity difference of SPR.Based on this principle, surface plasmon resonance biosensor is that part is fixed in metallic film surface with a kind of molecule of tool specific recognition attribute usually, the analyte in the monitoring solution and the cohesive process of this part.In compound formation or dissociation process, the refractive index of metallic film surface solution changes, and is detected by surface plasmon resonance biosensor immediately.
Compare with traditional interaction technology such as ultracentrifugation, fluorescence method, calorimetry etc., surface plasmon resonance biosensor has following distinguishing feature: (1) is detected in real time, dynamically the overall process of monitoring bio interaction of molecules; (2) need not the mark sample, kept molecular activity; (3) the sample requirement is few, and a general surface only needs about 1 μ g protein ligands; (4) testing process is convenient and swift, and is highly sensitive; (5) range of application is very extensive; (6) high flux, high-quality analysis data; The stability of (7) can tracing and monitoring fixing part; (8) quantitative measurement of the compound balance of disturbance reponse not; (9) in most cases, do not need sample is carried out pre-service; (10) since SPR based on to not penetrating the catoptrical measurement of sample, can in sample muddiness even opaque, carry out so detect.Based on above characteristics, surface plasmon resonance biosensor is able in field widespread uses such as bio-molecular interaction, drug screening, clinical diagnosis, food detection and environmental monitoring, membrane biologies.
The present method that the detection and the evaluation of pathogenic microorganism are typically used separation and Culture combining form characteristic, biochemical identification, immunoassay; there are shortcomings such as complicated operation, specificity are not strong, required time is long in cultivation; and many microbe growth require high; biochemical character, antibiotic sensitive type isophenous feature instability are vulnerable to gene regulation, plasmid and obtain and lose and the influence of aspects such as technical operation.And based on the authentication method of molecular biology such as plasmid analysis, nucleic acid hybridization, pvuii restriction fragment analysis, PCR etc., can overcome with the microorganism phenotypic characteristic is the shortcoming and the influence factor of basic detection method, shorten detection time greatly, improved detection efficiency.But these technology still exist shortcomings such as complicated operation, the high and low flux of false positive rate, poor specificity in various degree.Thereby these technology have been limited clinical, especially the application in the lowered in field environment.The inspection and the evaluation of pathogenic microorganism during China present field work, what still adopt is conventional art and method, test item is few, length consuming time, sensitivity is low, and some also need just can carry out in special laboratory, and this obviously can not satisfy the needs of modern medicine rescue.
The utility model content
In view of this, the purpose of this utility model provides a kind of pathogenic microorganism device for fast detecting, can overcome shortcomings such as the high and low flux of complicated operation, false positive rate that prior art exists, poor specificity, and test item is various, the inspection of pathogenic microorganism and evaluation when being specially adapted to field work.
Pathogenic microorganism device for fast detecting of the present utility model, comprise surface plasma body resonant vibration bio-sensing pick-up unit, incident light passage, emergent light passage and intellectual analysis processing platform, described incident light passage comprises the adjustable polarized light source that cruises, and the described adjustable polarized light source that cruises comprises polarized light source and the light-source control system that shakes;
Described emergent light passage comprises convergent lens and photodetector, and described photodetector is arranged on the focus of convergent lens, and the output terminal of described photodetector and intellectual analysis processing platform link;
Further, described surface plasma body resonant vibration bio-sensing pick-up unit comprises surface plasma resonance biosensor and adjustable detection light source: described surface plasma resonance biosensor comprises reaction tank, lens I and matrix, described reaction tank is arranged on surface plasma resonance biosensor top, described lens I is arranged on the surface plasma resonance biosensor bottom, described matrix is arranged between reaction tank and the lens I, described matrix is provided with metal film, described metal film and reaction tank keep in touch, and are coated with a plurality of bioprobes on the described reaction tank in advance;
Further, described intellectual analysis processing platform comprises signal amplification circuit, A/D change-over circuit, be the signal analysis/treatment circuit and the display circuit of core with the microprocessor, described signal through the photodetector collection is sent into signal analysis/treatment circuit successively and is carried out analyzing and processing behind signal amplification circuit, A/D change-over circuit, the gained result sends into display device by display circuit and shows;
Further, described polarized light source comprises optical transmitting set, lens II, optical filter, polaroid and cylindrical lens, and the light that described optical transmitting set sends behind scioptics II, optical filter and the polaroid, is assembled by cylindrical lens generation angle successively, directive lens I bottom;
Further, the polarized light source control system comprises stepper motor, segmentation drive unit and controllor for step-by-step motor, the pulse signal that described controllor for step-by-step motor sends is converted into the angular displacement of stepper motor by the segmentation drive unit, controllor for step-by-step motor is electrically connected with stepper motor by the segmentation drive unit, by the frequency of control step pulse signal, can be to the accurate speed governing of stepper motor; The number of control step pulse can accurately be located stepper motor;
Further, also comprise temperature control equipment, described temperature control equipment is by microprocessor and peripheral circuit thereof, temperature measurement circuit, signal amplification circuit, the A/D change-over circuit, display interface circuit, temperature-adjusting circuit, constant temperature overload alarm circuit is formed, described temperature measurement circuit comprises temperature sensor and peripheral components thereof, described temperature sensor is arranged on the position of energy detection reaction pond temperature, the signal of gathering through temperature sensor is successively through signal amplification circuit, send into microprocessor behind the A/D change-over circuit and carry out analyzing and processing, described display interface circuit, temperature-adjusting circuit, constant temperature overload alarm circuit is electrically connected with microprocessor;
Further, described metal film is the structure of the outer gold-plated film of chromium film;
Further, described surface plasma body resonant vibration bio-sensing pick-up unit also comprises sample inlet and sample liquid outlet, and described sample inlet and sample liquid outlet are arranged on the top of reaction tank;
Further, described matrix is glass or silicon chip, and described lens I is spherical lens or triangular lens;
Further, described bioprobe comprises staphylococcus aureus bioprobe, pseudomonas aeruginosa bioprobe, clostridium tetani bioprobe, Bacillus perfringens bioprobe, shigella dysenteriae bioprobe, bacillus anthracis bioprobe and/or comma bacillus bioprobe.
Principle of work of the present utility model is: when solution to be measured contacts with surface plasmon resonance biosensor, each biomolecule of its biologic single molecular layer combines with target molecule specificity in the testing sample solution, form compound, the surface structure of biologic single molecular layer is changed, this variation can cause each biomolecule on this unimolecular layer the change of SPR phenomenon to occur, can detect the variation at SPR angle by pick-up unit.
The utility model realizes that the working method of a plurality of biological signals parallel detections (concentration) is: 1. sampling: prepare the number duplicate samples, this sample contains the target molecule of the concentration known that can combine with the bioprobe specificity of pre-bag quilt, and the concentration of target molecule increases progressively successively in the number duplicate samples, by the variation of different t constantly of detection system record of the present utility model and biomolecule SPR angle ω, make ω/t curve according to the relation of t and ω; 2. draw typical curve:, can obtain when reaction reaches balance the SPR balance angle ω of each biomolecule by ω/t curve
0Value, make the typical curve of this angle value concentration of target molecules C corresponding, promptly corresponding to the ω of each target molecule with this point
0/ C curve, this corresponding relation deposit in the intellectualized analysis platform of detection system, form database; 3. detect: when the concentration solution of detection position, obtain ω/t curve according to the SPR angle ω of each point and the time relationship of t earlier, obtain the ω of each SPR by ω/t curve
0Value is again by ω
0The corresponding relation of/C curve can obtain the concentration value of this target molecule.
The beneficial effects of the utility model are:
(1) detects by selecting pathogen, and design corresponding bioprobe, be arranged in the reaction tank, realized multi-channel detection, simplified testing process, improved detection efficiency;
(2) can overcome non-specific response preferably, realize biomolecule to be measured and interactional detection thereof have been improved sensor performance on the whole;
(3) changed the set-up mode of conventional light source, setting by the adjustable polarized light source that cruises, can be according to detecting needs, at any time adjust the incident angle of incident light, overcome traditional pick-up unit and caused the low shortcoming of detection efficiency, can fully satisfy the detection needs owing to having fixed optimized incidence;
(4) of the present utility model based on miniaturization Design, compact conformation has not only improved the thermal stability and the mechanical stability of instrument simultaneously.
The accurately mutual effect (multifactorial effect) of multiple material in the analytical specimen of the utility model, the differences between batches of having avoided single mensuration to bring; And greatly reduced detection time, quickened testing process; Easy to use, the detection sensitivity height can be used for a plurality of fields, particularly can fully satisfy the use needs of field environment, has broad application prospects.
Other advantages of the present utility model, target and feature will be set forth to a certain extent in the following description, and to a certain extent, based on being conspicuous to those skilled in the art, perhaps can from practice of the present utility model, obtain instruction to investigating hereinafter.Target of the present utility model and other advantages can realize and obtain by following instructions and claims.
Description of drawings
In order to make the purpose of this utility model, technical scheme and advantage clearer, below in conjunction with accompanying drawing the utility model is described in further detail, wherein:
Fig. 1 is a structural representation of the present utility model;
Fig. 2 is provided with synoptic diagram for bioprobe;
Fig. 3 transmits synoptic diagram for signal;
Fig. 4 is the control structure synoptic diagram of adjustable light source of cruising.
Embodiment
1-reaction tank; 2-lens I; 3-matrix; 4-metal film; 5-bioprobe set-point; 6-polarized light source; 7-polarized light source control system; 8-convergent lens; 9-photodetector; 10-signal amplification circuit; 11-A/D change-over circuit, 12-signal analysis/treatment circuit; 13-display circuit; 14-optical transmitting set; 15-lens II; 16-optical filter; 17-polaroid; 18-cylindrical lens; 19-temperature control equipment; 20-sample inlet; 21-sample liquid outlet; 22-intellectualized analysis platform; 23-stepper motor; 24-segmentation drive unit; 25-controllor for step-by-step motor.
Fig. 1 is a structural representation of the present utility model; Fig. 2 is that the bioprobe bag is by synoptic diagram; Fig. 3 transmits synoptic diagram (wherein dotted arrow indicates light signal and transmits circuit, and solid arrow is represented the electrical signal transfer circuit) for photosignal; Fig. 4 is the control structure synoptic diagram of adjustable light source of cruising.As shown in the figure, the utility model comprises SPR bio-sensing pick-up unit, incident light passage, emergent light passage and intellectual analysis processing platform 22:
Wherein, SPR bio-sensing pick-up unit comprises surface plasmon resonance biosensor: surface plasmon resonance biosensor comprises reaction tank 1, lens I2 and matrix 3, reaction tank 1 is arranged on surface plasmon resonance biosensor top, lens I2 is arranged on the surface plasmon resonance biosensor bottom, matrix 3 is arranged between reaction tank 1 and the lens I2, matrix 3 is provided with metal film 4, and metal film 4 keeps in touch with reaction tank 1, is provided with a plurality of bioprobe set-points 5 in the reaction tank 1;
The incident light passage comprises the adjustable polarized light source that cruises, and the adjustable polarized light source that cruises comprises polarized light source 6 and polarized light source control system 7; Polarized light source 6 comprises optical transmitting set 14, lens II15, optical filter 16 and polaroid 17, the light that optical transmitting set 14 sends is successively behind scioptics II15, optical filter 16 and the polaroid 17, through cylindrical lens 18 angle taking place assembles, directive lens I2 bottom, wherein, optical filter 16 can improve the monochromaticity of incident light, and polaroid 17 is adjustable polaroid, in order to produce satisfactory P polarized light.Optical transmitting set 14 in the present embodiment adopts energy consumption low, luminous stable led light source,
Polarized light source control system 7 comprises stepper motor 23, segmentation drive unit 24 and controllor for step-by-step motor 25, the pulse signal that controllor for step-by-step motor 25 sends is converted into the angular displacement of stepper motor 23 by segmentation drive unit 24, by the frequency of control step pulse signal, can be to stepper motor 23 accurate speed governing; The number of control step pulse can be to stepper motor 23 accurate positioning control.Its adjusting can drive 24 devices and control by the segmentation of centre, and this parameter is a variable element.Control program in the present embodiment is set to according to the requirement of embodiment: 0.1 second/step, the mobile accuracy in per step was 1mm.
The emergent light passage comprises convergent lens 8 and photodetector 9, and photodetector 9 is arranged on the focus of convergent lens 8; Photodetector 9 in the present embodiment adopts the area array CCD of precision height, stable performance.
Intellectual analysis processing platform 22 comprises signal amplification circuit 10, A/D change-over circuit 11, be the signal analysis/treatment circuit 12 and the display circuit 13 of core with the microprocessor, send into signal analysis/treatment circuit 12 successively through the signal of photodetector 9 collections behind signal amplification circuit 10, A/D change-over circuit 11 and carry out analyzing and processing, the gained result shows by display circuit 13 input and display devices.
Present embodiment also is provided with temperature control equipment 19, temperature control equipment 19 links with sample pool 1, and temperature control equipment 19 is made up of several parts such as single-chip microcomputer, temperature measurement circuit, A/D change-over circuit, display interface circuit, keystroke interface circuit, temperature-adjusting circuit, constant temperature overload alarm circuit and data storage circuitries.Temperature-adjusting circuit comprises controllable silicon and heater strip, temperature measurement circuit comprises temperature sensor and peripheral components, by the temperature sensor senses temperature variation, temperature signal is converted to frequency signal by the A/D change-over circuit, when being lower than or being higher than under the setting value, temperature prescribes a time limit, machine controllable silicon conducting or turn-off heater strip is controlled elevates the temperature or reduces.Memory circuit is to be used for storing the temperature and time that temperature control system carries out work, makes that data can not lost under the situation of system's power down.
In the present embodiment, metal film 4 plates one deck chromium earlier for using Vacuum Coating method on matrix, plate the structure of one deck gold film again on the chromium film, disturbs thereby can eliminate acoustic-electric effectively.
In the present embodiment, matrix 3 is selected glass substrate for use; Lens I2 is the triangular lens.
All optical elements all are arranged on one and boil in the black salable cylindrical shell, thereby reduce the influence of extraneous parasitic light, have improved accuracy of detection.
The experimental procedure of measuring the embodiment of multiple molecular conecentration in the test solution is as follows:
1. the bag quilt of bioprobe: feeding mol ratio in reaction tank is the biotinylated thiol derivant of 1:9 and the potpourri (PBS buffered environment) of OH root mercaptan dilution, react the sufficiently long time, form the self-assembled monolayer (SAM) that closely covers on each bioprobe set-point surface, with the flushing of PBS damping fluid, then use micro sample adding appliance again with staphylococcus aureus, pseudomonas aeruginosa, clostridium tetani, Bacillus perfringens, shigella dysenteriae, bacillus anthracis, the bioprobe of seven kinds of pathogens of comma bacillus injects each bioprobe set-point of reaction tank;
2. measure: when wrapping in the reaction tank, testing sample is fed reaction tank, regulate the angle of prism, the black line that resonance takes place is fallen within the sensing range of photodetector by after the bioprobe.Begin to detect, write down the spr signal of each points simultaneously by photodetector 9, send into intellectualized analysis platform and carry out analyzing and processing, the course of work of intellectualized analysis platform is as follows: obtain the ω/t curve of each point by spr signal, obtained the SPR equilibrium angle ω of each point again by the ω/t curve of each point
0, by with database in the ω of storage
0/ C curve data are compared, thereby obtain the concentration value of each point.
Utilize the move mode and the position of polarized light source controller control polarized light source, can be implemented in each position and guarantee all incident angle incident with the best, guarantee each index consistency, can realize the synchronous detection of multiple index, be particularly suitable for genomic functional analysis.
Explanation is at last, above embodiment is only unrestricted in order to the explanation the technical solution of the utility model, although the utility model is had been described in detail with reference to preferred embodiment, those of ordinary skill in the art is to be understood that, can make amendment or be equal to replacement the technical solution of the utility model, and not breaking away from the aim and the scope of the technical program, it all should be encompassed in the middle of the claim scope of the present utility model.
Claims (10)
1. a pathogenic microorganism device for fast detecting comprises surface plasma body resonant vibration bio-sensing pick-up unit, incident light passage, emergent light passage and intellectual analysis processing platform, it is characterized in that:
Described incident light passage comprises the adjustable polarized light source that cruises, and the described adjustable polarized light source that cruises comprises polarized light source (6) and polarized light source control system (7);
Described emergent light passage comprises convergent lens (8) and photodetector (9), and described photodetector (9) is arranged on the focus of convergent lens (8), and the output terminal and the intellectual analysis processing platform of described photodetector (9) link.
2. pathogenic microorganism device for fast detecting according to claim 1, it is characterized in that: described surface plasma body resonant vibration bio-sensing pick-up unit comprises surface plasma resonance biosensor, described surface plasma resonance biosensor comprises reaction tank (1), lens I (2) and matrix (3), described reaction tank (1) is arranged on surface plasma resonance biosensor top, described lens I (2) is arranged on the surface plasma resonance biosensor bottom, described matrix (3) is arranged between reaction tank (1) and the lens I (2), described matrix (3) is provided with metal film (4), described metal film (4) keeps in touch with reaction tank (1), is coated with a plurality of bioprobes (5) on the described reaction tank (1).
3. pathogenic microorganism device for fast detecting according to claim 1 and 2, it is characterized in that: described intellectual analysis processing platform (22) comprises signal amplification circuit (10), A/D change-over circuit (11), display interface circuit (13) and be the signal analysis/treatment circuit (12) of core with the microprocessor, the described signal of gathering through photodetector (9) is successively through signal amplification circuit (10), send into signal analysis/treatment circuit (12) behind the A/D change-over circuit (11) and carry out analyzing and processing, described display interface circuit (13) is electrically connected with signal analysis/treatment circuit (12).
4. pathogenic microorganism device for fast detecting according to claim 3, it is characterized in that: described polarized light source (6) comprises optical transmitting set (14), lens II (15), optical filter (16), polaroid (17) and cylindrical lens (18), the light that described optical transmitting set (14) sends is successively behind scioptics II (15), optical filter (16) and the polaroid (17), by cylindrical lens (18) angle takes place and assemble, directive lens I (2) bottom.
5. pathogenic microorganism device for fast detecting according to claim 4, it is characterized in that: described polarized light source control system (7) comprises stepper motor (23), segmentation drive unit (24) and controllor for step-by-step motor (25), described controllor for step-by-step motor (25) is electrically connected with stepper motor (23) by segmentation drive unit (24), and the pulse signal that described controllor for step-by-step motor (25) sends is converted into the angular displacement of stepper motor (23) by segmentation drive unit (24).
6. pathogenic microorganism device for fast detecting according to claim 5, it is characterized in that: also comprise temperature control equipment (19), described temperature control equipment (19) is by microprocessor and peripheral circuit thereof, temperature measurement circuit, signal amplification circuit, the A/D change-over circuit, display interface circuit, temperature-adjusting circuit, constant temperature overload alarm circuit is formed, described temperature measurement circuit comprises temperature sensor and peripheral components, described temperature sensor is arranged on the position of energy detection reaction pond temperature, the signal of gathering through temperature sensor is successively through signal amplification circuit, send into microprocessor behind the A/D change-over circuit and carry out analyzing and processing, described display interface circuit, temperature-adjusting circuit, constant temperature overload alarm circuit is electrically connected with microprocessor.
7. according to the pathogenic microorganism device for fast detecting of claim 6, it is characterized in that: described metal film (4) is the structure of the outer gold-plated film of chromium film.
8. pathogenic microorganism device for fast detecting according to claim 7, it is characterized in that: described surface plasma body resonant vibration bio-sensing pick-up unit also comprises sample inlet (20) and sample liquid outlet (21), and described sample inlet (20) and sample liquid outlet (21) are arranged on the top of reaction tank (1).
9. according to claim 2 or 5 or 6 or 7 or 8 described pathogenic microorganism device for fast detecting, it is characterized in that: described matrix (3) is glass or silicon chip, and described lens I (2) is spherical lens or triangular lens.
10. pathogenic microorganism device for fast detecting according to claim 1 is characterized in that: described bioprobe comprises staphylococcus aureus bioprobe, pseudomonas aeruginosa bioprobe, clostridium tetani bioprobe, Bacillus perfringens bioprobe, shigella dysenteriae bioprobe, bacillus anthracis bioprobe and/or comma bacillus bioprobe.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200820099973 CN201262615Y (en) | 2008-09-12 | 2008-09-12 | Rapid detection device for pathogenic microorganism |
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| CN 200820099973 CN201262615Y (en) | 2008-09-12 | 2008-09-12 | Rapid detection device for pathogenic microorganism |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101349643B (en) * | 2008-08-18 | 2011-07-06 | 中国人民解放军第三军医大学第一附属医院 | Multichannel surface plasma wave sensing detection system |
| CN102253014A (en) * | 2011-04-15 | 2011-11-23 | 深圳大学 | System and method for surface plasmon resonance sensing detection |
| CN105486665A (en) * | 2016-01-26 | 2016-04-13 | 深圳大学 | SPR detection system and method |
| CN109813685A (en) * | 2019-02-21 | 2019-05-28 | 中国科学技术大学 | A kind of single bacteria surface plasma mechanical oscillation imaging method measuring waste water acute toxicity |
-
2008
- 2008-09-12 CN CN 200820099973 patent/CN201262615Y/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101349643B (en) * | 2008-08-18 | 2011-07-06 | 中国人民解放军第三军医大学第一附属医院 | Multichannel surface plasma wave sensing detection system |
| CN102253014A (en) * | 2011-04-15 | 2011-11-23 | 深圳大学 | System and method for surface plasmon resonance sensing detection |
| CN105486665A (en) * | 2016-01-26 | 2016-04-13 | 深圳大学 | SPR detection system and method |
| CN105486665B (en) * | 2016-01-26 | 2018-07-31 | 深圳大学 | A kind of SPR detection methods |
| CN109813685A (en) * | 2019-02-21 | 2019-05-28 | 中国科学技术大学 | A kind of single bacteria surface plasma mechanical oscillation imaging method measuring waste water acute toxicity |
| CN109813685B (en) * | 2019-02-21 | 2020-05-19 | 中国科学技术大学 | Single bacterium surface plasma mechanical vibration imaging method for determining acute toxicity of wastewater |
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Granted publication date: 20090624 Termination date: 20110912 |