CN107807115A - Metal covers light fluid Composition analyzed device - Google Patents

Abstract

A kind of metal covers light fluid Composition analyzed device, including：The test chamber being arranged on substrate and the sample intake passage and sample output passage that are attached thereto, test chamber top and bottom are respectively equipped with the glassy layer with metal film；Test chamber is surrounded for glass seal, and the glass seal or so is respectively equipped with through hole, and as sample intake passage and sample output passage, testing sample is sent into by sample intake passage, and waste sample is discharged by sample output passage；The aqueous solution or gas are provided with described test chamber.The present invention is formed using free-space coupling technology has high power density, the structure of high-quality-factor and highly sensitive three characteristics, it is high with photocontrol, stability, small volume, refractive index change hypersensitive, fast response time, the efficiency for strengthening spontaneous radiation, it can integrate, the advantages that damage threshold is high.

Description

Metal-covered optofluidic drug efficacy detector

technical field

The invention relates to a technology in the field of chemical detection, in particular to a real-time, non-labeled and highly sensitive metal-covered optofluid drug effect detector for detecting the action efficiency of anticancer drugs.

Background technique

Optofluidic biodetector is a micro-device for integrated biological sample analysis, which has the advantages of high integration, small size, high throughput, and low consumption. Since the basic optical material of the optofluidic device is liquid, the optical components based on the microfluidic optical detection chip can be easily realized through the fluid, and have a very natural compatibility with other components of the microfluidic chip. It is based on these advantages of optofluidic devices, using microfluidic structures to realize a new method of controlling light at the micron scale, and using these optofluidic devices combined with other components of microfluidics to realize the control of fluorescent molecules and biomolecules at the micron scale And other optical physical quantity detection research. It plays an important role in the field of biological detection. Real-time label-free, miniaturized, easy-to-drive, high-speed, and integrated photofluidic biochip detectors are such key devices as independent functional units. There is an urgent need in these fields. demand.

Contents of the invention

The present invention aims at the defect that the existing technology cannot monitor the interaction between the drug and the cancer cell in real time, and cannot detect the change of the fluorescence intensity after the interaction between the EGFR protein and the drug in the cancer cell in real time. Free space coupling technology forms a structure with three characteristics of high power density, high quality factor and high sensitivity, with light control, high stability, small size, ultra-sensitive to refractive index changes, fast response speed, and enhanced spontaneous emission efficiency , can be integrated, and the damage threshold is high.

The present invention is achieved through the following technical solutions:

The invention relates to a metal-covered optofluid drug effect detector, comprising: a detection cavity arranged on a loading body and a sample inlet channel and a sample output channel connected thereto, wherein: the top and bottom of the detection cavity are respectively equipped with metal The glass layer of the membrane.

The detection chamber is preferably surrounded by a glass gasket, and the glass gasket is provided with through holes on the left and right sides, serving as a sample inlet channel and a sample outlet channel, through which samples to be tested are fed in, and waste samples are discharged through the sample outlet channel.

Said detection chamber is provided with aqueous solution or gas, and the refractive index of the gas is preferably close to 1.0.

The inlet of the sampling channel is provided with a Y-shaped PDMS liquid injection mixing channel and a micro-injection pump for controlling the injection flow rate.

The outlet of the sample outlet channel is provided with a hose that directly leads to the waste liquid collector.

Between the glass layer and the carrier, between the glass gasket and the carrier and the glass layer are preferably optical adhesive bonding

The metal film is deposited on the surface of the glass layer by evaporation technology, and the thicknesses of the metal film on the glass layer at the top and bottom of the detection chamber are respectively calculated according to different incident light.

The sum of the thicknesses of the glass layer and the glass gasket is the thickness of the optofluid; the thickness of the glass gasket is the thickness of the detection cavity.

The Q quality factor of the metal-covered photofluid detector is analyzed: Q～λ/Δλ, wherein: λ is the wavelength of incident light, and Δλ is the phase difference value of the wavelength of light coupled into the photofluid, that is, the difference value of the mode. Because the high-order guided mode is generated in the optofluidic cavity, the highest order of the mode reaches ~2000, so Δλ~1/2000, so the Q quality inside the metal optofluidic can therefore reach ~10 ⁴ .

The invention relates to a detection method based on the metal-covered photofluidic detector. The parallel light emitted by the laser is incident on the upper surface of the metal-covered photofluidic detector; when the incident light passes through the metal layer, it is coupled into the detector through free space, And the light coupled into the cavity undergoes multiple oscillations and coherence to form a stable high-order mode standing wave field, generating reflected light and fluorescence signals containing black absorption lines due to attenuated total reflection (ATR), and the reflected light is transmitted through the beam splitter The light and fluorescence signals are decomposed into two beams, which are respectively collected and detected by the linear array CCD and high-resolution camera to measure the proportion of fluorescence in the optofluid, so as to obtain the changes of molecules in the detection cavity, and accurately calculate the amount of drug needed to treat cancer .

The relationship between the coupling angle θ _ATR of the free space coupling and the intrinsic loss Im(β ⁰ ) of the metal film and the radiation loss Im(Δβ ^L ) satisfies: Among them: ε ₂ is the refractive index of air, β ^L = β ⁰ + Δβ ^L , k ₀ is the propagation constant of light propagating in the air; therefore, the reflectivity corresponding to different angles is also different, corresponding to The relationship of reflectivity R satisfies: Therefore, it can be obtained that R _min ∼0 when (Im(β ⁰ )=Im(Δβ ^L )).

The excitation efficiency of the fluorescent signal, that is, the spontaneous emission rate

Among them: ρ _c (v)[ρ _f (v)] are the photon number densities with and without the cavity, respectively, ΔP is the full-height half-width of the power density distribution in the cavity, and Δρ _c is the full-height half-width of an intracavity mode . According to the theory, it can be obtained that the efficiency of spontaneous emission enhancement of the entire metal-covered optofluidic detector microcavity is approximately

The specific steps of the accurate calculation include:

1) Real-time detection of the movement of the black line corresponding to the ATR peak on the linear array CCD until the black line stops moving, and the time T required for the reaction can be accurately obtained, and the reaction rate V= can be obtained according to the relationship between the moving distance S and the unit time S/unit time, judging the effect of drug binding to the target protein and the time of action from the reaction rate can be accurately judged, and the starting point of the time when the drug enters the body and the time period of action can be obtained.

2) Use a high-resolution camera to collect and count the proportions of fluorescence of different colors in the coupling area, that is, under different reaction times, the proportions of the same fluorescence that are counted separately change until the proportion of fluorescence no longer changes ; The whole process can reflect the efficiency of the interaction between the drug and the target protein without labeling, that is, the drug effect E=the amount of drug used (the change in the fluorescent area ratio corresponding to the drug) A*the fluorescent area ratio of the target protein Variation B.

3) The amount of drug Y=E*X required to obtain the content of the target protein through the efficiency of the interaction between the drug and the target protein, and the amount of drug used is accurate to micrograms.

technical effect

Compared with the prior art, the present invention has the following advantages:

1) The excitation light in the reflection part is attenuated due to the resonance of the guided mode, and the fluorescence signal with a longer wavelength deviates from the reflection angle output;

2) The laser beam is enlarged, and the change of the black line ATR line can be clearly observed on the line array CCD;

3) Metal-covered optofluidic detectors are used as the detection cavity and resonant cavity, which have high power density, high quality factor and high sensitivity;

4) Using the guided mode with effective refractive index N→0 as the probe, the sample can be either liquid or gas;

5) The structure is simple, the operation is convenient and the cost is low.

Description of drawings

Fig. 1 is the side-view structure schematic diagram of the optofluidic detector that the present invention proposes;

Fig. 2 is the structural representation of the photofluidic detector that the present invention proposes;

Fig. 3 is a schematic diagram of the packaging structure of the optofluidic detector proposed by the present invention;

In the figure: upper metal film 1, flat glass 2, detection chamber 3, glass gasket 4, lower metal film 5, glass substrate 6, sampling channel 7, sampling channel 8, Y-shaped sampling channel 9, 10, PDMS Carrier 11 , metal-covered optofluid detector chip 12 , laser 13 , beam amplifier 14 , linear array CCD detector 15 , beam splitter 16 , and high-resolution camera 17 .

Detailed ways

As shown in Figure 3, this embodiment includes: a metal-covered optofluidic detector A and a photoelectric detection module B, wherein: the metal-covered optofluidic detector structure A includes: an upper metal film 1, a flat glass 2, a detection chamber 3, Glass gasket 4, lower metal film 5, glass substrate 6, sampling channel 7, sampling channel 8, Y-shaped sampling channel 9, 10, PDMS carrier 11.

The upper metal film 1 is deposited on the surface of the flat glass 2 , and the lower metal film 5 is deposited on the bottom surface of the glass substrate 6 .

In order to ensure the parallelism of the waveguide cavity, the glass gasket 4 requires high parallelism, and the flat glass 2, the glass gasket 4 and the glass substrate 6 must be assembled together by optical glue technology.

The sum of the thicknesses of the flat glass 2 , the glass gasket 4 and the glass substrate 6 is the thickness of the waveguide layer, and the thickness of the glass gasket 4 is the thickness of the detection cavity 3 .

Two through holes are opened on both sides of the glass gasket 4, which respectively pass through the sample inlet channel 7 and the sample outlet channel 8 after the Y-shaped channel (9) 10 of the metal-covered optofluidic detector structure A is mixed. The sample inlet channel 7 enters the sample pool, while waste samples are discharged through the sample outlet channel 8 .

The upper metal film 1 and the lower metal film 5 are both made of silver, with thicknesses of 35nm and 200nm respectively; the dielectric coefficient is ε ₁ =ε ₅ =-18231+i0416.

The material of the flat glass 2 is optical glass with a thickness of 300 μm.

The material of the glass gasket 4 is optical glass, and its thickness is about 500 μm.

The material of the glass substrate 5 is optical glass with a thickness of 100 μm.

A sampling channel 7 and a sampling channel 8 are arranged on the glass substrate 6 .

The photoelectric detection module B includes: a laser 13 , a beam amplifier 14 , a linear array CCD detector 15 , a beam splitter 16 , and a high-resolution camera 17 .

The output wavelength of the laser 13 is 6328nm, and the divergence angle is required to be less than 0.01mrad.

This embodiment is based on the detection method of the above-mentioned device. Firstly, the parallel light emitted by the laser 13 is incident on the surface of the hollow metal-clad waveguide structure A at a certain angle. Since the laser has a certain line width and divergence angle, through pre-design, it can ensure that an ultra-high-order guided mode is excited near the incident angle, that is, the reflected light contains an attenuated total reflection absorption peak of the guided mode, so the excitation wavelength in the reflected light The light intensity of the light is greatly weakened, and the fluorescence with a longer wavelength is output in a direction different from the reflection angle. The position of the black line in the reflected light spot is reflected by the linear array CCD detector 15, and the high-resolution camera detection 17 detects the fluorescent light spot in the coupling area.

Real-time detection of the movement of the black line corresponding to the ATR peak on the linear CCD until the black line stops moving, and the time T required for the reaction can be accurately obtained; from 0 to T, any time can be based on whether the black line moves, and According to the relationship between the moving distance S and the unit time, the reaction rate V=S/unit time can be obtained. From the reaction rate, the effect of the combination of the drug with the target protein and the time of action can be accurately judged, and it can be concluded that the drug reacts when it enters the body. The starting point of time and the time period of action.

There are different colors of fluorescence in the coupling area of 0.1mm*0.1mm, and the image is collected by a high-resolution camera, and then the proportion of different colors of fluorescence in 0.1mm*0.1mm is counted by matlab software, the reaction time is different, and the statistics are the same The proportion of fluorescence occupation changes until the proportion of fluorescence occupation does not change; the whole process can reflect the efficiency of the interaction between the drug and the target protein without labeling, that is, the drug effect E = drug usage (the fluorescence corresponding to the drug The variation of the area occupancy ratio) A* the variation B of the fluorescence area occupancy ratio of the target protein.

The amount of drug Y=E*X required to obtain a certain amount of X target protein content through the interaction efficiency of the drug and the target protein; at the same time, because the occupancy ratio of the fluorescent area can be accurate to 0.001mm ² , the drug usage can be calculated Quantities are accurate to micrograms.

The above specific implementation can be partially adjusted in different ways by those skilled in the art without departing from the principle and purpose of the present invention. The scope of protection of the present invention is subject to the claims and is not limited by the above specific implementation. Each implementation within the scope is bound by the invention.

Claims (9)

1. A metal-covered optofluid drug effect detector, characterized in that it comprises: a detection chamber arranged on the substrate and a sample inlet channel and a sample outlet channel connected thereto, wherein: the top and bottom of the detection chamber are respectively provided with Glass layer with metal film; The detection chamber is surrounded by a glass gasket, and the glass gasket is provided with through holes on the left and right sides, which are used as a sample inlet channel and a sample outlet channel. The sample to be tested is sent in through the sample inlet channel, and the waste sample is discharged through the sample outlet channel; The detection chamber is provided with aqueous solution or gas. 2. metal-covered photofluidic drug efficacy detector according to claim 1, is characterized in that, the entrance of described sampling channel is provided with Y font PDMS liquid injection mixing channel and the micro-injection pump for controlling injection flow rate; The outlet of the sample outlet channel is provided with a hose and a peristaltic pump. 3. The metal-covered optofluid drug efficacy detector according to claim 1, characterized in that, between the glass layer and the substrate, between the glass gasket and the substrate and the glass layer are photoresists bonding. 4. The metal-covered optofluidic drug efficacy detector according to claim 1, characterized in that, the metal film is deposited on the surface of the glass layer by evaporation technology, and the metal on the glass layer at the top and bottom of the detection cavity The thickness of the film is calculated according to different incident lights; the sum of the thicknesses of the glass layer and the glass gasket is the thickness of the optical fluid; the thickness of the glass gasket is the thickness of the detection cavity. 5. The metal-covered optofluidic drug efficacy detector according to claim 1, characterized in that, the Q quality factor of the metal-covered optofluidic detector is analyzed: Q～λ/Δλ, wherein: λ is incident light Wavelength, Δλ is the phase difference value of the wavelength of the light coupled into the optical fluid, that is, the difference value of the mode; because the optical fluid cavity produces a high-order guided mode, the highest order of the mode reaches ~ 2000, so Δλ ~ 1/2000, so the metal The Q quality inside the optofluidic can therefore reach ~10 ⁴ . 6. A detection method based on the metal-covered optofluidic drug efficacy detector according to any one of the above claims, characterized in that, the parallel light emitted by the laser is incident on the upper surface of the metal-coated optofluidic detector; The metal layer is coupled into the detector through free space, and the light coupled into the cavity undergoes multiple oscillations and coherence to form a stable high-order mode standing wave field, resulting in reflected light and fluorescence containing black absorption lines that appear due to attenuated total reflection Signal, through the beam splitter, the reflected light and the fluorescence signal are decomposed into two beams, which are collected and detected by the linear array CCD and high-resolution camera respectively, and the measurement of the fluorescence occupancy ratio in the optofluid is obtained to obtain the change of molecules in the detection cavity, and accurately measure and calculate The amount of medicine needed to treat cancer. 7. The method according to claim 6, wherein the relationship between the coupling angle θ _ATR of the free space coupling and the intrinsic loss Im(β ⁰ ) of the metal film and the radiation loss Im(Δβ ^L ) satisfies: Among them: ε ₂ is the refractive index of air, β ^L = β ⁰ + Δβ ^L , k ₀ is the propagation constant of light propagating in the air; therefore, the reflectivity corresponding to different angles is also different, corresponding to The relationship of reflectivity R satisfies: Therefore, it can be obtained that R _min ∼0 when (Im(β ⁰ )=Im(Δβ ^L )). 8. The method according to claim 6, characterized in that the excitation efficiency of the fluorescent signal, i.e. the spontaneous emission rate Among them: ρ _c (v)[ρ _f (v)] are the photon number densities with and without the cavity, respectively, ΔP is the full-height half-width of the power density distribution in the cavity, and Δρ _c is the full-height half-width of an intracavity mode , the efficiency of spontaneous emission enhancement of the entire metal-covered optofluidic detector microcavity is approximately 9. The method according to claim 6, characterized in that, the specific steps of accurate calculation include: 1) Real-time detection of the movement of the black line corresponding to the ATR peak on the linear array CCD until the black line stops moving, and the time T required for the reaction can be accurately obtained, and the reaction rate V= can be obtained according to the relationship between the moving distance S and the unit time S/unit time, judging the binding effect of the drug and the target protein from the reaction rate and the time of action can be accurately judged, and the starting point of the time when the drug enters the body and the time period of action can be obtained; 2) Use a high-resolution camera to collect and count the proportions of fluorescence of different colors in the coupling area, that is, under different reaction times, the proportions of the same fluorescence that are counted separately change until the proportion of fluorescence no longer changes ; The whole process can reflect the efficiency of the interaction between the drug and the target protein without labeling, that is, the drug effect E=the amount of drug used (the change in the fluorescent area ratio corresponding to the drug) A*the fluorescent area ratio of the target protein Variation B; 3) The drug dose Y=E*X required to obtain the target protein content through the interaction efficiency of the drug and the target protein.