CN109490202A - A kind of immunoturbidimetry protein analyzer based on light miniflow laser - Google Patents
A kind of immunoturbidimetry protein analyzer based on light miniflow laser Download PDFInfo
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- CN109490202A CN109490202A CN201811329949.0A CN201811329949A CN109490202A CN 109490202 A CN109490202 A CN 109490202A CN 201811329949 A CN201811329949 A CN 201811329949A CN 109490202 A CN109490202 A CN 109490202A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/39—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using tunable lasers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/39—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using tunable lasers
- G01N2021/391—Intracavity sample
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/39—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using tunable lasers
- G01N2021/392—Measuring reradiation, e.g. fluorescence, backscatter
- G01N2021/393—Measuring reradiation, e.g. fluorescence, backscatter and using a spectral variation of the interaction of the laser beam and the sample
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Abstract
The invention discloses a kind of immunoturbidimetry protein analyzers based on light miniflow laser, belong to sensor technical field.The analyzer includes pump light source module, sample detection module and signal collection processing module.Optical parametric oscillator exports pulse pump light in pump light source module, and concentrated lens are assembled, and is incident on method amber resonant cavity using reflecting mirror.Mechanics amber resonant cavity is arranged in upper reflector and lower mirror parallel in sample detection module, and after pulse pump light is incident in method amber resonant cavity, stimulated radiation occurs for dye molecule, generates laser signal output.After signal collection processing module handles laser signal, processing result is output to computer.The present invention by light miniflow laser technology in conjunction with immunoturbidimetry, using light miniflow laser in method amber resonant cavity multiple reflections, increase the effect number between laser and compound, improve the sensitivity of protein detection；Required test sample amount is few, can be measured to various immunoglobulin contents.
Description
Technical field
The invention belongs to sensor technical fields, and in particular to a kind of immunoturbidimetry protein based on light miniflow laser point
Analyzer.
Technical background
Immunoturbidimetry is widely used to the numerous areas such as biomedicine, food hygiene, environmental monitoring.Traditional is immune
Turbidimetry is mainly to utilize the specific binding of antigen-antibody, under the action of strong electrolyte, forms the biggish compound of partial size.
In the case where antibody excess, the forming amount of compound and the concentration of antigen are directly proportional.Light and compound interact, it may occur that
Scattering is absorbed and is reflected, and transmitted light or scattered light intensity relationship proportional to determined antigen concentration.This method has time-consuming
The advantages that short, disposable, homogeneous reaction, but the disadvantages of sensitivity is lower, amount of samples is big, limits its application range.
Light microfluidic technology is integrating for optical technology (sensing, manipulate, imaging) and micro-fluidic system.Microchannel is placed in two
It may make up a Fa-Po cavity between piece reflecting mirror.Laser dye in channel generates stimulated radiation under the action of pump light.
Stimulated radiation photon roundtrip between the mirror of two sides constantly enhances, and ultimately forms laser output.Sample to be tested is formed
Compound is mixed with dyestuff, and the concentration of compound generates different degrees of loss to the laser in laser cavity, to influence
Export the intensity of laser.Therefore, this detection method has very high sensitivity.Traditional immunization protein analyzer mostly uses detection
The mode that light once penetrates cuvette is detected, and sample and reagent dosage are big.It is compared with the traditional method, utilization method of the present invention
The highly sensitive characteristic of amber chamber light miniflow laser proposes a kind of new immunoturbidimetry protein analyzer, utilizes miniflow laser
In intracavitary multiple reflections, the detection performance of traditional immunization turbidimetry can be significantly improved.Meanwhile the concentration by changing dyestuff,
Solvent or type can realize the measurement of different proteins concentration.
Summary of the invention
The present invention is for the faultiness design mentioned in background technique and has made a kind of immune ratio based on light miniflow laser
Turbid protein analyzer.The protein analyzer has the characteristics that structure is simple, measurement accuracy is high.
A kind of immunoturbidimetry protein analyzer based on light miniflow laser, including pump light source module 1, sample detection module
2 and 3 three parts of signal collection processing module.
The pump light source module includes: optical parametric oscillator (1), convergent lens (5), reflecting mirror (6), the optical parameter
Oscillator (1) exports pulse pump light, and concentrated lens (5) are assembled, the normal using reflecting mirror (6), with upper reflector (8)
Method amber resonant cavity is incident at 13-17 degree angle in direction.
The sample detection module includes: microchannel (7), upper reflector (8), support construction (9), lower reflecting mirror
(10), mechanics amber resonant cavity is arranged in parallel in the upper reflector (8) and lower reflecting mirror (10), and it is anti-to provide light for miniflow laser
Feedback, microchannel (7) and support construction (9) are set in method amber resonant cavity；The pulse pump light is incident on method amber resonant cavity
After interior, dye molecule absorbs pulse pump light energy, from ground state transition to excitation state, when a large amount of dye molecules are all in excitation state
When, it may occur that stimulated radiation occurs for population inversion, dye molecule, generates laser signal output.
The signal collection processing module includes: gathering element (11), optical fiber (12), spectrometer (13), computer (14),
The laser signal being collected into is coupled in optical fiber (12) by the gathering element (11), and laser signal is transferred to light along optical fiber (12)
Spectrometer (13), spectrometer (13) handle laser signal, and processing result is output to computer (14).
Further, the pump light source module further includes aperture (2), transmissibility of adjustable attenuation piece (3), energy probe (4).
The pulse pump light of optical parametric oscillator (1) output is adjusted after aperture (2) by transmissibility of adjustable attenuation piece (3), and
It is popped one's head in (4) by energy, then concentrated lens (5) are assembled.The aperture (2) can carry out space filtering to pulse pump light
And beam shaping.The energy of pulse pump light can be adjusted in the transmissibility of adjustable attenuation piece (3).The energy probe (4) can be right
The energy of pulse pump light carries out real-time monitoring.
Further, the reflectivity of the upper and lower reflecting mirror is above 90%, and the metal film or property of low cost can be used
The excellent deielectric-coating of energy, improves reflectivity, further promotes detectivity.
Further, the adjustable range of optical parametric oscillator optical maser wavelength is 189-2500nm.It can be according to the type of dyestuff
The pulse laser of different wave length is selected to be pumped, common pulse laser wavelength optional 355nm, 532nm, 635nm, 808nm,
1064nm etc..
Further, the microchannel (7) is made of rectangular quartz capillary and hose, outside rectangular quartz capillary
Diameter is 1.4mm × 1.4mm, and internal diameter is 1.1mm × 1.1mm.It is solid using ultraviolet glue between rectangular quartz capillary and plastic flexible pipe
Change connection.Microchannel can be used that wall thickness is thinner, and the higher material of transmitance further decreases the loss in method amber resonant cavity,
And then improve detection sensitivity.Rectangular capillary, the metal with microchannel (7) with size can be used in the support construction (9)
Block etc., to enhance the stability of method amber resonant cavity.
The present invention is by light miniflow laser technology in conjunction with immunoturbidimetry.On the one hand, antigen-antibody under given conditions can
It specifically binds, and forms the biggish compound of partial size under strong electrolyte effect.When antibody excess, formation it is compound
Object concentration is directly proportional to antigen concentration.On the other hand, light miniflow laser technology generally uses pulse pump light source as pump light
Source, dyestuff is passed into resonant cavity as gain media can be achieved laser output, by changing the wavelength of pump light, dyestuff
Type and the size of Fa Po resonant cavity can realize the laser signal output of different wave length.The compound and dye that antigen-antibody is formed
Material mixes, and is passed into method amber resonant cavity together, and the miniflow laser of generation can enhance light and compound phase in intracavitary multiple reflections
Interaction.When compound partial size is much smaller than miniflow optical maser wavelength, Rayleigh scattering can be generated, miniflow laser intensity can occur significantly
It reduces.And the degree of the intensity decline of miniflow laser is directly proportional to the concentration of compound, therefore can be swashed by detection miniflow
The intensity of light realizes the sensing of testing protein concentration.
The invention has the benefit that
1, the present invention utilizes light miniflow laser multiple reflections in method amber resonant cavity, increases between laser and compound
Number is acted on, the sensitivity of protein detection can be improved.
2, IgG density measurement method high sensitivity provided by the invention, easy to operate, required test sample amount
It is few, various immunoglobulin contents can be measured.
3, the type of dyestuff and the wave of pulse pump light can be changed in IgG density measurement method provided by the invention
Long, to realize the laser output of different wave length, the particle size of the compound formed according to testing protein selects different
Dyestuff and pulse pump light, to realize the analysis of different types of protein concentration.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of smooth miniflow laser immunoturbidimetry protein analyzer provided by the invention
Fig. 2 is the sectional view of smooth miniflow laser immunoturbidimetry protein analyzer provided by the invention
Fig. 3 is antigen-antibody reaction schematic diagram
Fig. 4 is the laser intensity and protein concentration of smooth miniflow laser immunoturbidimetry protein analyzer provided by the invention
Relation curve
Specific implementation method
A kind of production and application method of the immunoturbidimetry protein analyzer based on light miniflow laser, specifically include following
Step:
The immunoturbidimetry protein analyzer of the light miniflow laser of the present embodiment includes pump light source module 1, sample detection
3 three parts of module 2 and signal collection processing module.The pump light source module includes: 532nm pulse laser (1), assembles
Lens (5), reflecting mirror (6), the 532nm pulse laser (1) exports pulse pump light, using aperture (2) to pulse
Pump light carries out space filtering and beam shaping, pulse pump light energy is adjusted by transmissibility of adjustable attenuation piece (3), and by energy
Amount probe (4) carries out real-time monitoring, and then concentrated lens (5) are assembled, the method using reflecting mirror (6), with upper reflector (8)
Method amber resonant cavity is incident at 15 degree of angles in line direction；Meanwhile gathering element (11) position being mixed up and being fixed, then use optical fiber
(12) it is connect with spectrometer (13), so that the output signal of spectrometer is transmitted to computer (14).
The sample detection module includes: the upper reflector (8) that reflectivity is 91.5%, reflectivity be 99.5% it is lower anti-
It penetrates mirror (10), reflection membrane material is deielectric-coating.Microchannel (7) includes the circle of rectangular quartz capillary and two sides connection
Rectangular capillary both ends are connected by plastic flexible pipe with round plastic hose, and junction carries out solidification filling using ultraviolet glue.It will be micro-
Circulation road (7) and support construction (9) are put into togerther between two reflecting mirrors, mechanics amber resonant cavity.Method amber resonance is adjusted simultaneously
The position of chamber, so that convergent beam is incident on just in method amber resonant cavity.
Standard items are diluted according to certain concentration gradient, take the dilute sample of 10 μ L and the R1 reagent of 1125 μ L
(Kanggong department, Erie, immunoglobulin G detection reagent box) is uniformly mixed, and is placed in 37 DEG C of water-bath, and heated at constant temperature is incubated for
5min.The R2 reagent (Kanggong department, Erie, immunoglobulin G detection reagent box) of 375 μ L is added into above-mentioned mixed solution again simultaneously
It is uniformly mixed, is placed in 37 DEG C of water-bath, heated at constant temperature is incubated for 5min.Take 900 μ L reaction complete solution in another from
In heart pipe, the rhodamine B aqueous solution that 100 μ L concentration are 1.2mM is added thereto, is uniformly mixed.
The solution mixed is passed through in microchannel (7), as shown in Figure 2.Pulse pump light source is opened, difference is recorded
Laser intensity corresponding to the standard items of concentration.Every time when measurement, observation energy probe (4) registration, while adjusting adjustable damping
Piece (3), to ensure that each pump energy is consistent.The standard items of various concentration are drawn out into a mark with corresponding laser intensity again
Directrix curve.
For sample to be tested, above-mentioned measuring process is repeated, the laser intensity measured is compared with standard curve
Obtain the specific concentration of sample.
As shown in figure 3, antigen-antibody is specifically bound, and under the action of strong electrolyte, through after a period of time it
Afterwards, partial size biggish compound is formed under the action of strong electrolyte.Microchannel (7) is placed between two reflecting mirrors can structure
At a Fa-Po cavity.Laser dye in channel generates stimulated radiation under the action of pump light.Stimulated radiation photon is on two sides
Roundtrip between mirror constantly enhances, and ultimately forms laser signal output.By sample to be tested formed compound and dyestuff into
Row mixing, complex concentration generate different degrees of loss to the laser in laser cavity, to influence to export the strong of laser signal
Degree.When pump energy is fixed on 35.37uJ/mm2When, the standard concentration of experimental calibration and the relation curve of laser intensity are as schemed
Shown in 4.
Corresponding dyestuff and pumping wavelength can be selected for different testing proteins.Such as the IgM of Siemens Company's production
Dyestuff can be changed into HITC in 800nm or so by kit, the compound partial size formed, and pumping wavelength is selected
600nm.Therefore, the detection of IgM concentration can also be completed on this device.
Claims (6)
1. a kind of immunoturbidimetry protein analyzer based on light miniflow laser, including pump light source module 1, sample detection module 2
With 3 three parts of signal collection processing module；
The pump light source module includes: optical parametric oscillator (1), convergent lens (5), reflecting mirror (6), the optical parametric oscillation
Device (1) exports pulse pump light, and concentrated lens (5) are assembled, the normal direction using reflecting mirror (6), with upper reflector (8)
Method amber resonant cavity is incident at 13-17 degree angle；
The sample detection module includes: microchannel (7), upper reflector (8), support construction (9), lower reflecting mirror (10), institute
It states upper reflector (8) and mechanics amber resonant cavity is arranged in parallel in lower reflecting mirror (10), provide light feedback for miniflow laser, miniflow is logical
Road (7) and support construction (9) are set in method amber resonant cavity；After the pulse pump light is incident in method amber resonant cavity, dyestuff
From ground state transition to excitation state population inversion occurs for molecule absorption pulse pump light energy, and dye molecule is excited spoke
It penetrates, generates laser signal output；
The signal collection processing module includes: gathering element (11), optical fiber (12), spectrometer (13), computer (14), described
The laser signal being collected into is coupled in optical fiber (12) by gathering element (11), and laser signal is transferred to spectrometer along optical fiber (12)
(13), spectrometer (13) handles laser signal, and processing result is output to computer (14).
2. a kind of immunoturbidimetry protein analyzer based on light miniflow laser as described in claim 1, it is characterised in that: described
Pump light source module further includes aperture (2), transmissibility of adjustable attenuation piece (3), energy probe (4)；The optical parametric oscillator (1) is defeated
Pulse pump light out is adjusted after aperture (2) by transmissibility of adjustable attenuation piece (3), and is popped one's head in (4) by energy, then through meeting
Poly- lens (5) are assembled；The aperture (2) carries out space filtering and beam shaping to pulse pump light；The adjustable damping
The energy of pulse pump light can be adjusted in piece (3)；The energy probe (4) supervises the energy of pulse pump light in real time
It surveys.
3. a kind of immunoturbidimetry protein analyzer based on light miniflow laser as claimed in claim 1 or 2, it is characterised in that:
The reflectivity of the upper and lower reflecting mirror is above 90%, using metal film or deielectric-coating.
4. a kind of immunoturbidimetry protein analyzer based on light miniflow laser as described in claim 1, it is characterised in that: described
The adjustable range of optical parametric oscillator optical maser wavelength is 189-2500nm.
5. a kind of immunoturbidimetry protein analyzer based on light miniflow laser as described in claim 1, it is characterised in that: described
Microchannel (7) is constituted by rectangular quartz capillary with its both ends using the hose that ultraviolet adhesive curing is connected to.
6. a kind of immunoturbidimetry protein analyzer based on light miniflow laser as described in claim 1, it is characterised in that: described
Support construction (9) is with microchannel (7) with the rectangular capillary or metal block of size.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111172232A (en) * | 2020-01-09 | 2020-05-19 | 电子科技大学 | Optical fiber micro-flow laser sensor based on nano particles |
| CN113075133A (en) * | 2021-03-01 | 2021-07-06 | 电子科技大学 | Particle-enhanced immunoturbidimetric protein analyzer based on optical microfluidic laser |
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| CN106404729A (en) * | 2016-08-26 | 2017-02-15 | 电子科技大学 | Manufacturing and application method of ion sensor based on optofluidic bio-laser |
| CN107589275A (en) * | 2017-08-02 | 2018-01-16 | 电子科技大学 | A kind of flow-velocity sensing method and device based on light miniflow dye laser |
| CN108398388A (en) * | 2018-01-16 | 2018-08-14 | 电子科技大学 | A kind of integrated array colorimetric analysis device based on one-dimension optical-fiber micro-flow laser |
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2018
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106404729A (en) * | 2016-08-26 | 2017-02-15 | 电子科技大学 | Manufacturing and application method of ion sensor based on optofluidic bio-laser |
| CN107589275A (en) * | 2017-08-02 | 2018-01-16 | 电子科技大学 | A kind of flow-velocity sensing method and device based on light miniflow dye laser |
| CN108398388A (en) * | 2018-01-16 | 2018-08-14 | 电子科技大学 | A kind of integrated array colorimetric analysis device based on one-dimension optical-fiber micro-flow laser |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111172232A (en) * | 2020-01-09 | 2020-05-19 | 电子科技大学 | Optical fiber micro-flow laser sensor based on nano particles |
| CN111172232B (en) * | 2020-01-09 | 2023-02-28 | 电子科技大学 | Optical fiber micro-flow laser sensor based on nano particles |
| CN113075133A (en) * | 2021-03-01 | 2021-07-06 | 电子科技大学 | Particle-enhanced immunoturbidimetric protein analyzer based on optical microfluidic laser |
| CN113075133B (en) * | 2021-03-01 | 2022-10-14 | 电子科技大学 | Particle-enhanced immunoturbidimetric protein analyzer based on optical microfluidic laser |
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