KR20110133778A - Optoelectrofluidic immunoassay platform and method thereof - Google Patents

Abstract

PURPOSE: An optoelelctrofluidic immunoassay platform and an immunoassay method are provided to enable antigen detection of high sensitivity. CONSTITUTION: An optoelectrofluidic immunoassay platform comprises: a light source(200) for irradiating light; a pattern forming device(300) which is placed on the light pathway and changes light pattern; a optoelectrofluidic device(100) for irradiating light pattern and injecting a sample; and a power supply device(700) for applying power to the optoelelctrofluidic device. The immunoassay platform further comprises a condensing lens(400) and light source(500) for detection.

Description

Optoelectronic fluid control immunoassay device and method {OPTOELECTROFLUIDIC IMMUNOASSAY PLATFORM AND METHOD THEREOF}

The present invention relates to an immunoassay apparatus and method using the photoelectron fluid control technology, and more particularly, the reaction and concentration of the antigen and antibody in a small amount of fluid sample using the electrokinetic principle induced by light And a photoelectrofluid controlled immunoassay device and method for detecting antigens quickly and simply by controlling washing.

Immunoassays using specific antigen-antibody responses are widely used in the fields of chemical analysis and clinical diagnostics to detect specific substances in biological samples. Immunoassay methods include Radio Immunoassay (RIA), Enzyme Immunoassay (EIA), and Fluorescence Immunoassay (FIA), depending on the type of label used for detection. Because of its high sensitivity, it is one of the most widely used methods. These immunoassays are usually required to undergo antigen-antibody reactions and then to separate the reactants and non-reactants so that the amount of reactants can be accurately detected. Therefore, automated equipment has been commercialized and widely used to conveniently perform such repeated reaction, separation, washing and detection processes [A. Ford, "Automated immunoassay analyzers Survey," CAP TODAY, Colleage of American Pathologists, June 2008, pp. 24-74].

However, in the case of the automated equipment, the configuration of a plurality of chambers, holders, frames, etc. is very complicated, the manufacturing cost is also very high, and requires a sample amount of several hundred microliters or more.

In order to overcome this limitation, an immunoassay device using a microfluidic device has also been developed [X. Jiang, JMK Ng, AD Stroock, SKW Dertinger, and GM Whitesides, J. Am . Chem . Soc . 125, pp. 5294-5295 (2003). An immunoassay device using a microfluidic device can rapidly analyze several samples simultaneously and can analyze even samples of several hundred microliters or less.However, a pump and a tube for injecting a sample are always required. Since the amount of sample discarded is very large compared to the amount, the amount of sample actually used is not small compared to the automated equipment. In addition, the microfluidic device used once has a limitation in that it cannot be reused, consuming a large number of devices and tubes.

Therefore, the first problem to be solved by the present invention is to overcome the limitations of the existing immunoassay device, and it is possible to quickly perform an automated immunoassay in small sample fluid droplets of several hundred nanoliters or less without complicated additional equipment. It is to provide a photoelectrofluid controlled immunoassay device for that.

The second problem to be solved by the present invention is a photoelectrofluid controlled immunoassay method that can quickly perform automated immunoassay in small sample fluid droplets of several hundred nanoliters or less, and can simultaneously perform a plurality of analytes. To provide.

In accordance with another aspect of the present invention, there is provided a photoelectrofluid controlled immunoassay device according to the present invention, wherein a light source irradiates light and a pattern is formed to change the pattern of the light by being located in a path of light irradiated from the light source. Optoelectronic fluid control immunoassay device including an apparatus, an opto-electronic fluid device is irradiated with a light pattern passing through the pattern forming device and a sample containing the material to be analyzed is injected, and a power supply device for supplying power to the opto-electronic fluid device It may include.

The photoelectrofluid controlled immunoassay device according to the present invention may include one or more condensing lenses for condensing light output from the pattern forming device between the pattern forming device and the photoelectric fluid device.

The pattern forming apparatus includes a device driver that can be adjusted by a computer and is capable of automated image manipulation by a computer equipped with a device driver.

The pattern forming apparatus may be any one of a digital micromirror device (DMD) or a liquid crystal display (LCD).

The optoelectronic fluid element is positioned on a stage, and the stage may be provided with a motor for automation, and may move up, down, left, and right to change a light pattern region irradiated through the condenser lens.

The light source may be any one selected from a halogen lamp, a light emitting diode, and a laser.

The photoelectromagnetic fluid controlled immunoassay device according to the present invention may include a detection light source for irradiating incident light to the photoelectromagnetic fluid element in order to optically observe the sample.

It may include a signal detector for detecting a signal due to a material electronic state or a material vibration state generated by the incident light.

The signal detector may be any one of a CCD, a photodiode, a photomultiplier, and a spectrometer, and the signal detector includes a signal processor, and the signal processor processes and stores the received signal to generate data. do.

The photoelectromagnetic fluid controlled immunoassay device according to the present invention may include at least one focusing lens for focusing incident light irradiated from the light source for detection and an output signal output from the photoelectric fluid element.

The focusing lens for focusing the incident light and the output signal may be a plurality of different lenses.

The photoelectromagnetic fluid control immunoassay device of the present invention further comprises a focusing lens for focusing incident light output from the light source for detection and an output signal output from the photoelectric fluid element, and incident light output from the light source for detection. And further comprising a first mirror for irradiating the optoelectronic fluid element and the sample, and further comprising a second mirror for reflecting the output signal output from the optoelectronic fluid element to the signal detector. It is done.

The detection light source and the signal detector may further include a shutter for adjusting the reception time of the incident light and the output signal, wherein the shutter is opened after the immune reaction is terminated in the sample and the voltage is cut off.

The signal detector further includes a filter or a pinhole for filtering the signal output from the sample.

In the photoelectromagnetic fluid controlled immunoassay device of the present invention, the light source, the shutter, the stage, and the signal detector may be automatically adjusted by being synchronized by the computer-controlled control module.

The photovoltaic fluid element is positioned on a surface to which the light pattern is irradiated, and is located on a surface opposite to the photoconductive layer and a photoconductive layer in which current is conducted only to a region where the light pattern is irradiated by the light source and the light pattern forming apparatus. A space layer for forming an electric field in the sample from a voltage applied to the photoconductive layer, and a space layer formed between the photoconductive layer and the ground electrode layer and spaced apart from the photoconductive layer and the ground electrode layer; It may include. The space layer may include a sample in which an electric field is formed and an electrodynamic phenomenon occurs when voltage and light patterns are applied by the power supply device and the light pattern forming apparatus.

An injection hole for injecting the samples and a flow path through which the samples can move may be further provided.

The photoconductive layer may include a substrate, a plate electrode to which a voltage is applied through the power supply device, and a photoconductive material to which a current to selectively apply voltage is applied only to a region to which the light pattern is irradiated.

The plate electrode may be a transparent conductive material selected from gold, aluminum, copper, an N-type silicon substrate, and ITO.

The photoconductive material may be any one selected from hydrogenated intrinsic amorphous silicon, cadmium sulfide, and npn phototransistor.

The photoconductive layer further includes an intermediate layer doped between the photoconductive material and the plate electrode, wherein the intermediate layer reduces contact resistance between the photoconductive material and the electrode and maximizes the characteristics of the photoconductive layer.

The intermediate layer may be any one selected from amorphous silicon, aluminum and molybdenum.

A protective layer is provided between the photoconductive layer and the space layer to protect the photoconductive material, and the protective layer may be made of silicon nitride or silicon oxide.

The space layer further includes a microstructure or superhydrophobic material, wherein the microstructure or superhydrophobic material is formed between the photoconductive layer and the ground electrode layer to separate a plurality of samples.

The ground electrode layer may include a substrate and a plate electrode, and the plate electrode may be any one of a transparent conductive material selected from gold, aluminum, copper, an N-type silicon substrate, and ITO.

The transparency of the photoconductive layer and the ground electrode layer depends on the irradiation direction of the light pattern and the output direction of the output signal, and the layer exists in the irradiation direction of the light pattern, the output direction of the signal from the sample, or the direction in which the detector is arranged. The electrode is characterized in that made of a transparent conductive material.

The sample may include microparticles that serve as a support layer, analytes, and probe microparticles for measurement.

The microparticles serving as the support layer and the probe microparticles form an immunocomplex through the analyte.

The microparticles may be polymer microparticles that are polystyrene or latex, and the probe microparticles may be fluorescent nanoparticles or metal nanoparticles labeled with Raman scattering material.

The analyte may be one or more of antigen-antibody, DNA, RNA, and protein.

In the space layer, when the voltage is applied to the space layer and the light pattern is irradiated with the power supply device and the pattern forming device, an electric field is formed in the sample inside the space layer, and an electrodynamic phenomenon may occur.

The electrokinetic phenomenon may be any one of dielectric electrophoresis (DEP, dielectrophoresis), AC electroosmosis (ACEO) and electrophoresis.

The sample is characterized in that when the light is irradiated from the light source can output an output signal due to a material electronic state or a material vibration state.

The material electronic state is fluorescence emission or light absorption by wavelength, and the material vibration state is Raman scattering or IR absorption.

The signal detector is characterized by detecting a signal due to a material electronic state or a material vibration state.

The material electronic state is fluorescence emission or light absorption by wavelength, and the material vibration state is Raman scattering or IR absorption.

The output signal may be identified as an electronic signal by a CCD, a photodiode, a photomultiplier tube, a spectrometer, or the like, and may be detected using a confocal fluorescence microscope when performing simultaneous multiple detection using a single microparticle.

In accordance with another aspect of the present invention, an optoelectronic fluid controlled immunoassay method according to the present invention includes injecting samples into an optoelectronic fluid device, irradiating a light pattern to the optoelectronic fluid device, Applying a voltage, forming an electric field in the optoelectronic fluid device by the voltage and the irradiated light pattern, causing samples in the optoelectronic fluid device to flow to induce an antigen-antibody reaction, Moving and concentrating the antigen-antibody reacted samples into a light pattern irradiated region, and moving the probe microparticles and targets which have not reacted in the concentration step in a direction irradiated or opposite to the light pattern irradiated The amount of the probe microparticles combined with the microparticles or the support in the step of concentrating A step for irradiating an incident light beam for detection on the specimen to chulhagi, the light of the incident light and the sample - the signal by the material interaction are output, and may include the step of detecting the output signal to the detector.

The light pattern irradiating step is characterized by irradiating a light pattern on a portion of the optoelectronic fluid device.

In the reaction step and the concentration step, the electrophoresis generally occurs, or a frequency in the range of 1 kHz to 1 MHz in which both the electrophoresis and the electroosmotic electrophoresis occur.

In the washing step and the detection step, a frequency voltage in a range of 100 Hz to 1 kHz where alternating electroosmotic flow is performed for washing may be applied.

In the washing step and the detection step, a frequency voltage in the range of 10 kHz to 100 kHz may be additionally applied to reassemble or reconcentrate the fine particles.

The electron fluid controlled immunoassay method according to the present invention may simultaneously detect a plurality of antigens present in each sample by injecting a plurality of samples at the same time, the microstructure or a superhydrophobic material to separate the plurality of samples It can be formed in a space layer.

According to the photoelectrofluid controlled immunoassay device and method according to the present invention, a device having a conventional complex configuration is not required by controlling reaction, concentration, washing, and detection of analyte through a sample image that is automatically displayed by a program. The immunofluorescence analysis is performed by concentrating the sample using the electrofluidism caused by the material interaction, and thus, the automated fluorescence immunoassay can be performed using only a few hundred nanoliter samples and a simple photoelectron fluid control device.

The photoelectrofluid controlled immunoassay device and method according to the present invention is based on various optical measurement methods such as surface enhancement Raman scattering as well as fluorescence, and thus can detect high sensitivity antigens within minutes and analyze multiple samples at the same time. Can be performed at a time, there is an advantage that can control the reaction of the antibody-antigen freely.

1 is a conceptual diagram of an opto-fluid control immunoassay device according to the present invention.
2 is a cross-sectional view showing the structure and interior of the optoelectronic fluid device according to the present invention.
3 is a detailed conceptual diagram of an opto-fluid control immunoassay device according to the present invention.
Figure 4 is a flow chart showing the process of the photoelectrofluid control immunoassay method according to the present invention.
5 is a graph showing the form of intensity for each output signal wavelength detected according to the presence or absence of a target material.
FIG. 6 is an image illustrating a process in which the microparticles serving as the support are concentrated by a programmed image pattern in the photoelectron fluid controlled immunoassay method according to the present invention.
7A to 7C are results of analysis using fluorescence (red) nanoparticles as probe microparticles and using biotin-attached IgG (Immunoglobulin G) as a target material in the photoelectron controlled immunoassay method according to the present invention.
Figure 7a is an image showing the distribution of the microparticles and probe microparticles serving as a support layer using different fluorescent filters.
Figure 7b is an image and a graph showing the fluorescence brightness of the fine particles when washed.
Figure 7c is an image and a graph showing the fluorescence brightness of the microparticles not washed.
8a to 8b is a nanoparticle coated with a Raman probe molecule that outputs a specific Raman scattering signal in a photoelectrofluid controlled immunoassay method according to the present invention as a probe microparticle, and targets a cancer marker AFP (alpha fetoprotein) As a result of analysis using a substance,
Figure 8a is a graph showing the intensity of the Raman scattering signal according to the concentration of the target material.
FIG. 8B is a graph illustrating a specific peak signal (1615 cm ⁻¹ ) among the Raman scattering signals and comparing the concentration with the concentration of the target material.
FIG. 9 is a fluorescence image obtained by simultaneously capturing a large number of microparticles and simultaneously washing the probe nanoparticles which do not stick using the photoelectrofluid controlled immunoassay device according to the present invention.
FIG. 10 is a conceptual diagram illustrating a photoelectrofluid controlled immunoassay method using an antibody immobilized on an electrode surface of the ground electrode layer. FIG.
<Explanation of symbols for the main parts of the drawings>
100: photoelectric fluid element 110: photoconductive layer
111 substrate 112 electrode
113: photoconductive material 120: ground electrode layer
121 substrate 122 electrode
130: space layer 131: flow
140: immune complex 141: microparticles
142: material to be analyzed 143: probe microparticles
144 antibody immobilized on an electrode 200 light source
300: pattern forming apparatus 310: device driver
400: condenser lens 500: light source for detection
520: focusing lens 530: first mirror
540: second mirror 550: shutter
560: shutter 570: pinhole
600: signal detector 700: power supply device
800: computer 810: control module
900: stage A: light pattern
B: incident light C: output signal
(a): sample injection step (b): light pattern irradiation step
(c): voltage application step (d): reaction step
(e): concentration step (f): incident light irradiation step
(g): signal detection step

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. First, it should be noted that the same components or parts in the drawings represent the same reference numerals as much as possible. In describing the present invention, detailed descriptions of related well-known functions or configurations are omitted in order not to obscure the gist of the present invention.

1 is a conceptual diagram illustrating an apparatus for controlling photoelectron fluid control according to the present invention.

The photoelectromagnetic fluid control immunoassay device according to the present invention includes a light source 200, a pattern forming device 300, an optoelectronic fluid device 100, and a power supply device 700.

The light source 200 is for generating and controlling an electric field in the photovoltaic fluid device 100 to perform an immunoassay, and for manipulating a sample injected into the photovoltaic fluid device, and selecting one of a halogen lamp, a light emitting diode, and a laser. According to a preferred embodiment of the present invention, it is preferable that the wavelength range is in the visible light region.

The pattern forming apparatus 300 is positioned in a path of light irradiated from the light source 200 and changes a pattern of light irradiated from the light source, and is a digital micromirror device (DMD) and a liquid crystal display (LCD). , PDP, OLED may be any one selected from the display device, according to a preferred embodiment of the present invention, the pattern forming apparatus is preferably an LCD. The use of a flat panel display such as an LCD has the advantage that the system is simpler to configure and portable.

The light pattern changed by the pattern forming apparatus means that the light is irradiated at various points while the light irradiated from the light source corresponding to the back light of the LCD passes through the LCD module, and the light passes through the LCD module. It means that the light is changed to enable automated light manipulation through the program. When using a DMD, unlike LCD, light is formed by using the principle that light is selectively reflected in each area.

The light pattern A changed by the pattern forming apparatus 300 may be freely changed to form virtual electrode patterning, and may be automatically driven through a program, and light may be changed by the changed light pattern. The point to be irradiated can be formed singularly or plurally. In addition, it is characterized in that it is possible to perform a plurality of processes at the same time.

In one embodiment according to the present invention, the LCD is a light pattern using a 1024 × 768 pixel LCD (product number L3T57S) used in a projector called CP-S225 of HITACHI, but is not limited thereto.

The optoelectronic fluid device 100 will be described in detail with reference to FIG. 2. 2 is a cross-sectional view showing the structure and interior of the optoelectronic fluid device 100 according to the present invention.

The optoelectronic fluid device 100 includes a photoconductive layer 110, a ground electrode layer 120, and a space layer 130. If necessary, an intermediate layer (not shown), a protective layer (not shown), and a sample injection hole (not shown) are provided. C) and a microfluidic flow path (not shown).

According to a preferred embodiment of the present invention, there is no limitation on the shape of the photovoltaic fluid device 100, but it is preferable that the photovoltaic fluid device is in the form of a rectangular parallelepiped, and may be adjusted according to process and driving conditions, but the photoconductive The thickness of the layer and the ground electrode layer is 0.5 mm to 1 mm, and the height of the space layer is preferably 0.005 mm to 0.3 mm.

The photoconductive layer 110 is located on the surface to which the light pattern A is irradiated, and a current is conducted only in a region where the light pattern A is irradiated. The photoconductive layer includes a substrate 111 and an electrode ( 112, a photoconductive material 113.

According to a preferred embodiment of the present invention, it can be adjusted according to the process and driving conditions, but the thickness of the substrate is 0.35 mm to 0.7 mm, the thickness of the electrode is 10 nm to 1000 nm, the thickness of the photoconductive material is 800 nm It is preferable that it is to 3000 nm.

Voltage is applied to the electrode 111 through the power supply device. On the other hand, when the light pattern or the output signal is incident to the photoconductive material through the substrate or the electrode, the substrate is most preferably using a glass substrate, the electrode 112 is ITO (indium tin) It is preferably made of a transparent conductive material such as oxide, gold thin film.

The photoconductive material 113 has a characteristic of a phototransistor that forms an electric field in the sample by conducting a current only in a specific region to which the light pattern A is irradiated. Such a photoconductive material may be made of photoconductive materials such as hydrogenated intrinsic amorphous silicon, cadmium sulfide (CdS) or npn phototransistor.

In order to reduce the contact resistance between the photoconductive material and the electrode and maximize the characteristics of the photoconductive layer, an intermediate layer may be included between the photoconductive material and the electrode, wherein the intermediate layer is doped amorphous silicon. ), Aluminum or molybdenum.

In addition, a protective layer for protecting the photoconductive material may be located between the photoconductive layer and the space layer. The protective layer may be made of one of silicon nitride and silicon dioxide.

The ground electrode layer 120 is positioned on a surface opposite to the photoconductive layer and forms an electric field in the sample from a voltage applied to the photoconductive layer.

The ground electrode layer 120 includes a substrate 121 and an electrode 122, and a voltage is applied to the electrode 122 through the power supply device.

According to a preferred embodiment of the present invention, it can be adjusted according to the process and driving conditions, but the thickness of the substrate is 0.35 mm to 0.7 mm, the thickness of the electrode is preferably 10 nm to 1000 nm.

On the other hand, when the light pattern or the output signal (C) is incident to the photoconductive material through the substrate 121 or the electrode 122, the substrate 121 is most preferably using a glass substrate The electrode 122 is preferably made of a transparent conductive material such as indium tin oxide (ITO), a gold thin film, and the like.

On the other hand, the transparency of the photoconductive layer 110 and the ground electrode layer 120 depends on the irradiation direction of the light pattern (A) and the output direction of the sample signal, the output of the signal from the irradiation direction of the light pattern or the sample The electrode of the layer present in the direction or in the direction in which the signal detector 600 is disposed is preferably made of a transparent conductive material.

The space layer 130 is characterized in that the sample is injected, the electric field is formed when the voltage and the light pattern is applied, the electrodynamic phenomenon occurs.

The sample is composed of microparticles 141 serving as a supporting layer, an analyte 142, and probe microparticles 143 for measurement. In this case, the microparticles serving as the support layer and the probe microparticles may form an immune complex 140 through the target material.

Even when the amount of the sample injected into the space layer 130 is several hundred nanoliters or less, the antigen can be detected by the photoelectrofluid controlled immunoassay device of the present invention. According to a preferred embodiment of the present invention, the injection amount of the sample It is preferable that silver is 500 nL or more. The injection amount of the sample depends on the concentration of the analyte, the height of the space layer, the area of the light pattern, and the like. Even if the analyte is present in the sample very few to less than a few atomol, it is possible to concentrate the analyte and the probe microparticles using the electrodynamic phenomenon controlled by the light pattern, and the reaction time and The mixing method is also freely controllable. The concentration and mixing is preferably mixed and concentrated the sample using an electroosmotic flow generated under an alternating electric field of 10 kHz or less.

The sample may also be injected simultaneously with a plurality of samples to detect several kinds of antigens present in each sample at the same time. In this case, a microstructure (not shown) or a superhydrophobic material (not shown) may be formed between the photoconductive layer 110 and the ground electrode layer 120 to separate the plurality of samples. In addition, it may include an injection port (not shown) for injecting each sample, and may include a microfluidic flow path (not shown) for moving each sample.

The microparticles 141 may be polymer microparticles such as polystyrene and latex, and the target material 142 may be one or more of antigen-antibody, DNA, RNA, and protein, and fluorescent nanoparticles as the probe microparticles. And metal nanoparticles labeled with a material causing Raman scattering.

In the photoelectromagnetic fluid controlled immunoassay device according to the present invention, when a voltage is applied using the power supply device 700 and the light pattern A is irradiated, an electric field is formed in the space layer 130, and an electrodynamic phenomenon Will occur.

The electrodynamic effects include dielectrophoresis (DEP), AC electroosmosis (ACEO), electrophoresis and the like.

Dielectric behavior is a phenomenon in which a dielectric exhibits an electric dipole due to an electromagnetic induction phenomenon and is moved by force in a non-uniform electric field. The movement of these particles is determined by the difference in permittivity between the particles and the liquid around them, and the size of the particles (proportional to the cube of the radius) and the magnitude of the electric field gradient (proportional to the gradient of the square of the electric field). This will affect the speed of movement.

The dielectrophoresis includes negative dielectric movements in which the fine particles move in a direction in which the electric field is weak (the opposite direction to the direction of light irradiation) and positive movements of the fine particles in a direction in which the electric field is strong (the direction in which the light is irradiated). ) There is genetic activity. The nature of the electrophoresis may vary depending on the type of fluid, the type of microparticles and molecules, and the frequency of the AC voltage signal.

In the non-uniform electric field of the alternating electroosmotic, ions in the fluid form a thin electric double layer on the surface of the electrode and the liquid interface, and under the influence of a tangential electric field formed by voltage This is a phenomenon in which fluid moves along the surface.

The electroosmotic phenomenon causes a flow 131 in the direction in which the electric field is strong, that is, the direction in which the light pattern is irradiated, and generates a vortex at the edge of the light pattern. Induce. The concentration and dispersion characteristics of the electroosmosis depend on various physical and chemical properties such as the frequency of the AC voltage signal and the type, size, and amount of charge of the material or medium.

In addition, gravity force and electrostatic interactions can affect the behavior of the sample.

The electrostatic interaction is a force acting between two or more entities by electric charges induced in the dielectric under an electric field, and has characteristics of pushing or pulling each other. Pulling forces occur except when the two dielectric materials are located parallel to a plane perpendicular to the direction of the electric field. Like electrophoresis, electrostatic interactions may vary depending on the type of fluid, the type of microparticles and molecules, and the frequency of the AC voltage signal.

The various phenomena act in combination to affect the behavior of the sample. At this time, the behavior of the sample depends on the magnitude and frequency of the voltage applied from the power supply device and the light pattern. Thus, the microfluidic control device performs an immunoassay that undergoes various steps such as reaction, washing, and detection. There is a feature that can be controlled freely through the light pattern manipulation. Finally, the purpose of the present invention is to detect the type and amount of the target substance present in the sample by optically or electronically detecting the output signal from the immunoassay.

As shown in FIG. 2, when the photoconductive layer 110 is positioned on the ground electrode layer 120, in the case of fine particles of several micrometers or more, the light pattern is not irradiated due to the influence of negative dielectric and gravity. In the case of nanoparticles of 100 nanometers or less, they are hardly influenced by gravity or dielectric force, and collect in a light pattern direction by alternating electroosmotic flow. In addition, the metal nanoparticles or biomolecules are also affected by a weak amount of the electrophoretic force, so the light pattern is collected in the area irradiated.

Therefore, the microparticles 141, the analysis target material 142, and the probe microparticles 143 for the measurement are injected into the photovoltaic fluid device 100, and the voltage is applied to the light pattern after the support layer as the sample. When A) is irradiated, the light particles are collected in the unirradiated area of the microparticles, and the target material and the probe microparticles are washed away in the direction of the light pattern. Therefore, only the probe microparticles attached to the microparticles through the target material can be detected.

The microparticles may be polymer microparticles such as polystyrene and latex, and the target material may be one or more of protein and DNA. As the probe microparticles, fluorescent nanoparticles, metal nanoparticles labeled with a material causing Raman scattering, and the like may be used. In order to know the amount of the target material, the size of the fluorescence signal should be measured in the case of fluorescent nanoparticles, and in the case of the metal nanoparticles, the surface of the Raman signal of the material causing the Raman scattering attached to the nanoparticles is amplified by the metal nanoparticles. Surface-enhanced Raman scattering (SERS) signals should be measured. In this case, by using several kinds of probe microparticles, there is a characteristic that several kinds of target substances can be detected at the same time.

The power supply device 700 is a device for applying a voltage to the photoelectric fluid device 100, characterized in that the electric field is formed in the photoelectric fluid device by the voltage applied by the power supply device, the power supply device is AC power It may be a device or a DC power supply, and when using an AC power supply, the shape and offset voltage of the signal may be adjusted as well as the voltage magnitude of the frequency.

Figure 3 is a conceptual diagram showing in more detail the photoelectrofluid control immunoassay device according to the present invention.

According to a preferred embodiment of the present invention, the optoelectronic fluid controlled immunoassay device according to the present invention includes a light collecting lens 400 in the light source 200, the pattern forming apparatus 300, the optoelectronic fluid device 100 and the power supply 700. ) May further include a detection light source 500, a signal detector 600, a computer 800, and a stage 900.

A condenser lens 400 for condensing the light pattern 203 formed by the pattern forming apparatus 300 between the pattern forming apparatus 300 and the photovoltaic fluid element 100 on the photovoltaic fluid element 100 is further included. In addition, the pattern forming apparatus may be adjusted by the computer 800 through the device driver 310, thereby enabling automated image manipulation.

The condenser lens 400 may use a 10-fold, 20-fold, 40-fold, or 50-fold objective lens, and a plurality of such lenses may be provided, and the plurality of lenses may be different from each other so that the light pattern may be irradiated onto the optoelectronic fluid element. It can be configured as a lens.

The optoelectronic fluid device 100 is positioned on the stage 900, and the stage may further include a motor (not shown) for automation, and the stage is light irradiated through the condensing lens 400. It can be moved up, down, left and right so as to concentrate the sample by changing the pattern area.

In the photoelectron fluid controlled immunoassay device of the present invention, after an immunoassay is performed by the light pattern in the photoelectric fluid device, the detection light source 500 for optically observing the light pattern and the photo-material reaction of the sample is further included. It may include.

The immunoassay means that the microparticles, the probe microparticles and the antigen-antibody target material in the sample are photoreacted with the irradiated light pattern, and then the target material to which the microparticles are bound and the target material to which the microparticles are not bound are analyzed. It is a process.

The detection light source 500 outputs one or more laser lights having a white light or a specific wavelength distribution, or one or more optical choppers or pulsed lasers capable of intermittently or continuously irradiating light onto the sample. pulse laser). Such a light source irradiates the incident light (B) to the sample to output a signal due to a material electronic state such as fluorescence emission, light absorption for each wavelength, or Raman scattering, IR absorption, or the like. Characterized in that the output signal (C) due to the vibration state of the material can be output.

According to a preferred embodiment of the present invention, the detection light source 500 is preferably a laser having a wavelength range of 480 nm to 670 nm, or a halogen lamp having a filter in a specific region of the wavelength band.

The photoelectromagnetic fluid controlled immunoassay device of the present invention may further include a signal detector 600 for detecting a signal due to a material electronic state or a material vibration state generated by the incident light B.

The signal detector 600 detects a signal due to a material electronic state such as fluorescence emission or wavelength absorption of the sample, or a material vibration state such as Raman scattering or IR absorption. Detect signals by The signal detector 600 may be any one of a charge coupled device (CCD), a photodiode, a photomultiplier tube, and a spectrometer capable of wavelength analysis of light. The signal detector may further comprise a signal processor for processing and storing the received signal to generate data.

The photoelectrofluid controlled immunoassay device of the present invention includes a first focusing lens 520 and a first lens for irradiating incident light B for detection into the optoelectronic fluid element, and for transmitting an output signal C generated therefrom to the signal detector. The mirror 530 and the second mirror 540 may be further included.

And a first mirror 530 that reflects incident light so that the incident light B output from the detection light source 500 is irradiated on the photoelectric fluid element and the sample, and the output output from the sample. The second mirror 540 may further include a signal C to reflect the signal to the signal detector 600.

The focusing lens 520 focuses the incident light B output from the detection light source 500 onto the sample. The lens used in the present invention can be used 10 times, 20 times, 40 times, 50 times the objective lens, there can be a plurality of such lenses, the focusing lens is a lens for focusing the light output from the light source for detection and The lens for observing the sample may be composed of a plurality of different lenses.

The second mirror 540 is configured to reflect the output signal C outputted from the specimen to the signal detector 600. The second mirror may be configured by replacing with a beam splitter, and may be a dichromatic mirror.

The detection light source 500 may further include a shutter 550 for adjusting an irradiation time of incident light, and the signal detector 600 may further include a shutter for adjusting a reception time of the output signal.

According to a preferred embodiment of the present invention, it is preferable that the irradiation time of the incident light B of the detection light source is 10 msec or more, and the reception time of the output signal C of the signal detector is 10 msec or more. As the reception time of the output signal C of the signal detector is longer, the magnitude of the output signal is increased, and as a result, the detection sensitivity of the analyte is increased. When the incident light B of the detection light source is a laser, its intensity is preferably 100 W / cm ² or less, and when the intensity of the light source is too high and the irradiation time of the incident light B is too long, the locality of the sample is The change in temperature may cause flow due to the heat transfer effect. This heat transfer effect may allow the reaction and mixing to occur better, but it is more desirable that such temperature change does not occur due to the nature of the temperature-sensitive immunoassay.

In the photoelectromagnetic fluid controlled immunoassay device of the present invention, when the incident light B is irradiated to the photoelectromagnetic fluid element 100 while the power is applied, the voltage is cut off after the immune reaction is completed. Thereafter, the incident light B is irradiated onto the optoelectronic fluid element, and for this purpose, the incident light B is irradiated by the filter and the shutter 550 provided with the incident light B to the detection light source 500. Can be controlled.

The photoelectron fluid immunoassay device of the present invention converts the output signal (C) into the signal detector (C) through a filter or pinhole 570 additionally provided to the signal detector 600 and converts the signal into an electronic signal. The analysis results are shown.

The signal detector 600 may further include a shutter 560 to adjust a time at which an output signal is detected. The longer the shutter 560 remains open, the more the output signal is input to the signal detector, and the larger signal is measured.

The photoelectron fluid immunoassay device further includes the light source 200, the detection light source 500, the shutters 550, 560, the stage 900, the signal detector 600, and the like in the computer 800. It can be adjusted automatically by synchronizing through the provided control module 810.

One operation of the photoelectrofluid controlled immunoassay device according to the present invention is as follows.

The light is irradiated from the light source 200, the pattern is changed by the pattern forming apparatus 300 positioned in the path to which the light is irradiated, and the changed light pattern A passes through the condensing lens 400, and the condensing is performed. The light pattern A passing through the lens is irradiated to the photovoltaic fluid device 100 into which the sample containing the analyte is injected, and the stage 900 moves up and down vertically and horizontally to move the light to the photovoltaic fluid device. The light layer is irradiated to a part and / or the entirety of the optoelectronic fluid element 100 by moving the focal point and the irradiation area of the pattern, and when power is applied to the optoelectronic fluid element by the power supply device, a space layer into which the sample in the optoelectronic fluid element is injected. An electric field is formed at 130, and the light pattern and the sample react in the space layer in which the electric field is formed.

The sample includes microparticles, probe microparticles, and analyte, and the light pattern is irradiated and a voltage applied to a specific frequency is applied so that the electric field is formed in the area where the microparticles are collected in a region where the light pattern is not irradiated. In the case of the target material and the probe microparticles, an immunoassay is performed to wash away the light pattern.

After the immunoassay is performed, the power supply device is cut off to cut off the voltage applied to the photovoltaic fluid device, and the incident light B is irradiated onto the sample from the light source 500 for detection in order to optically observe the result of the immunoassay. The incident light, fluorescent nanoparticles, and probe microparticles labeled with a material causing Raman scattering react to output a signal due to a change in the material electronic state and the material vibration state, and the output signal C is transmitted by the signal detector 600. Detect. The irradiation time of the incident light B is adjusted by the shutter 550, and the reception time of the output signal C is adjusted by the shutter 560, and appears on the detector through a filter.

The light source, the pattern forming device, the stage, the power supply, the detection light source, the shutter, and the signal detector are synchronized by the control module and can be automatically adjusted by a computer, whereby the reaction, concentration, cleaning and Detection can be manipulated to detect analytes, such as antigens.

4 is a flow chart showing the process of the photoelectrofluid controlled immunoassay method according to the present invention. As shown in FIG. 4, the photoelectron fluid immunoassay method according to the present invention includes a sample injection step (a), a light pattern irradiation step (b), a voltage application step (c), a reaction step (d), and a concentration step (e). ), A washing step (not shown), a detection incident light irradiation step (f), and a signal detection step (g).

The photoelectrofluid controlled immunoassay method according to the present invention has been described above in the description of the apparatus. Here, the differences will be described and the same contents will be briefly described.

In the sample injection step (a), a sample including an analyte is injected through a sample injection hole (not shown) connected to the space layer of the optoelectronic fluid device.

In the light pattern irradiating step (b), the light pattern A whose pattern is changed by the pattern forming apparatus positioned in the path of light output from the light source is irradiated to the optoelectronic fluid element.

The light source may output one or more laser lights having a specific wavelength distribution, or may output visible light. The light pattern conducts a current to a specific region of the photovoltaic fluid element to form an electric field, and manipulates the light pattern to freely manipulate the shape of the electric field. In addition, the motion of the sample can be freely controlled using electrodynamic principles.

The light source irradiates the light pattern (A) to the sample to output a signal due to a material electronic state such as fluorescence emission, light absorption for each wavelength, or Raman scattering or IR absorption. It is possible to output a signal due to a vibration state of a substance or the like.

The light pattern (A) is characterized in that the pattern is changed by the pattern forming apparatus 300, the pattern forming apparatus is a liquid crystal display (LCD), a digital micromirror device (DMD), PDP, OLED or Aperture and the like may be used, and the pattern forming apparatus may include a condenser lens 400 for condensing a light pattern on the optoelectronic fluid element.

In the light pattern irradiation step, the position of the photovoltaic fluid element is moved by the stage 900 so that the light pattern is irradiated to a partial region of the light pattern A photovoltaic fluid element.

In the voltage applying step (c), a voltage is applied to the optoelectronic fluid device 100 by the power supply device 700. The power supply device may be an AC power supply device or a DC power supply device. When the AC power supply device is used, the shape and offset voltage of the signal may be adjusted as well as the voltage size of the frequency.

The reaction step (d) forms an electric field in the sample in the photovoltaic fluid device by the applied voltage and the irradiated light pattern, and causes the samples to flow to induce an antigen-antibody reaction.

The concentration step (e) is characterized in that to irradiate a light pattern on a portion of the optoelectronic fluid device, to move the region to which the light pattern is irradiated to collect the antigen-antibody reaction samples to the region where the light pattern is irradiated Concentration.

In the reaction step (d) and the concentration step (e), generally, electrophoresis occurs well, or a frequency in the range of 1 kHz to 1 MHz in which both the electrophoresis and the electroosmotic flow occur may be used.

The washing step (not shown) is a step of washing the probe microparticles and the target material less reacted by alternating osmotic flow in the direction in which the light pattern is irradiated or in the opposite direction.

The detecting incident light irradiation step (f) is a step of irradiating incident light onto a sample in order to optically and electronically observe and detect the amount of probe microparticles bound to the microparticles through an analyte.

The incident light is irradiated with a light pattern (A) to the sample to output a signal due to a material electronic state such as fluorescence emission, light absorption for each wavelength, or Raman scattering, IR absorption (infrared absorption) It is possible to output a signal due to a vibration state of a substance or the like.

In the washing and detecting step, a frequency of 100 Hz to 1 kHz in which AC electroosmotic flow occurs well may be used for washing, and at this time, a voltage of 10 kHz to 100 kHz is applied to assemble or reconcentrate the fine particles. May be

In one embodiment according to the present invention by applying a frequency of 1 kHz and moving the light pattern from the outside to the inside to promote the reaction of the microparticles and the target material, probe microparticles, and concentrated the microparticles. At this time, the microparticles concentrated inward away from the light pattern under the influence of negative dielectric force, alternating electroosmotic flow, and gravity. In addition, the mixing and reaction of the sample occur simultaneously by the alternating current electroosmotic flow. At this time, the target material and the probe microparticles which do not cause a reaction are washed by the alternating osmotic flow and go out in the direction of the light pattern. Thereafter, a voltage of 10 kHz was applied to the microparticles to be assembled by negative dielectric and electrostatic interactions to facilitate detection.

The signal detecting step (g) is a step in which a signal by photo-material interaction is output from the sample, and the signal is detected by a detector.

In the signal detection step, the probe microparticles and the analyte to which do not react move in the direction in which the light pattern is irradiated or vice versa, and a signal is output by the probe microparticles coupled through the analyte. Is detected by a detector.

In this case, if the amount of the probe microparticles is measured through the optical signal in the signal detecting step, the light pattern and the power supply for the manipulation are preferably turned off.

5 is a graph showing the shape of the intensity of each output signal wavelength detected according to the presence or absence of the target material in the photoelectron fluid control immunoassay device and method according to the present invention.

The output signal C may be a signal from the probe microparticles and may include fluorescence, Raman scattering, and the like.

When the target material is present, the probe microparticles are combined with the microparticles that become the support via the target material, and the output signal is detected according to the amount of probe microparticles that adhere to the support even after the washing process.

On the other hand, when the target material does not exist, the probe microparticles cannot bind to the microparticles that become the support, and thus are all washed and separated in the direction of the light pattern during the washing process. At this time, the output signal of the probe microparticles separated from the microparticles serving as the support is not measured.

As a result, the output signal varies depending on the amount of the target material, and the photoelectrofluid controlled immunoassay can quickly and accurately measure the amount and type of the target material.

Figure 6 is an embodiment according to the present invention, in the photoelectrofluid controlled immunoassay method according to the present invention is an image showing a process in which the microparticles as a support is concentrated by a programmed image pattern in chronological order.

When the 1 kHz frequency is applied, the fine particles spread out from the light pattern are concentrated in the center direction, and when the 10 kHz frequency is applied, they are assembled by electrostatic attraction and locally concentrated by negative dielectric force. You can check it. At this time, the probe microparticles much smaller than the microparticles are washed in the direction of the light pattern after the reaction with the microparticles.

Figure 7a to 7c is an embodiment according to the present invention, using fluorescent (red) nanoparticles as probe microparticles in the photoelectrofluid controlled immunoassay method according to the present invention, the biotin-attached IgG (Immunoglobulin G) target material The analysis results are shown as follows.

6 micrometer-sized fluorescent (green) microparticles were used as the support particles. The fluorescent nanoparticles are coated with Neutravidin to bind with the Biotin. In addition, the microparticles are coated with a monoclonal antibody to IgG to bind to the IgG. As a result, the fluorescent nanoparticles and the fine particles serving as the support may form an immunocomplex through the IgG as the target material.

Figure 7a is an image showing the distribution of the microparticles and probe microparticles serving as a support layer using different fluorescent filters. The part shown by the dotted white line shows the area | region which is not irradiated with light, ie, the area | region where the microparticles used as a support layer concentrate and detect an output signal. Therefore, the inside of the white dotted line is the washed area, the outside is the area that is not washed, the microparticles that serve as the support layer and the target material and the probe microparticles that do not form an immunocomplex are washed out.

7b and 7c are images and graphs showing the fluorescence brightness of the washed fine particles and the unwashed microparticles, respectively. In the case of microparticles not washed, the amount of probe fluorescent nanoparticles bound to the microparticles serving as the support layer cannot be properly measured, whereas in the case of washed microparticles, the probe fluorescence bound to the microparticles serving as the support layer can be measured. The amount of nanoparticles can be accurately measured.

8A to 8B illustrate, according to an embodiment of the present invention, a nanoparticle coated with a Raman probe molecule that outputs a specific Raman scattering signal as a probe microparticle, and an AFP (alpha fetoprotein) that is a cancer marker as a target material. The analysis results are shown. 6 micrometer-sized fluorescence (480 nm output) microparticles were used as the support microparticles. The silver nanoparticles are coated with a polyclonal antibody against AFP to bind with the AFP. In addition, the microparticles are coated with a monoclonal antibody against AFP to bind with the AFP. As a result, the nanoparticles and the microparticles serving as the support may form an immunocomplex through the AFP which is the target material. At this time, the amount of the target material attached to the microparticles serving as the support may be estimated through the magnitude of the Raman scattering signal output from the nanoparticles.

At this time, the Raman scattering is a phenomenon in which the photon energy (hv) having a specific frequency is scattered by the photon energy (hv ') of a different frequency while changing the vibration state of the molecule. Raman scattering at this time includes elastic Rayleigh scattering, Stokes Raman scattering and anti-Stokes Raman scattering.

The Raman scattering signal may be amplified by a surface-enhanced Raman scattering effect when the nanoparticle is a metal material.

This phenomenon is also associated with surface plasmon resonance at the metal surface. Surface plasmon refers to surface electromagnetic waves in which collective vibration of surface charges generated along the interface between metal and dielectric occurs when an electric field is applied to the metal surface forming the interface with the dielectric. Metals exhibiting this phenomenon are mainly used for materials such as gold, silver, copper, and aluminum that emit electrons easily and have a negative dielectric constant, and electromagnetic waves for applying an electric field are p-polarized surfaces. Due to the nature of the plasmon, TM polarized electromagnetic waves are used.

In general, when electromagnetic waves are incident on a metal material, they are totally reflected at the metal surface and the evanescent field decreases exponentially into the metal film at the interface. When the phases of the waves coincide, resonance occurs, all of the incident wave energy is absorbed by the metal film, and the reflected wave disappears, and the electric field distribution in the direction perpendicular to the boundary decreases exponentially away from the boundary. It is called surface plasmon resonance. In general, it is known that the surface enhancement effect of Raman scattering is maximized when light in the wavelength band where surface plasmon resonance is most likely to be incident.

The frequency at which surface plasmon resonance occurs is influenced by the assembly state of nanoparticles and microparticles. Therefore, by adjusting the frequency or magnitude of the AC voltage, the distance between the nanoparticles and the microparticles and the density of the assembly can be controlled. Through this, the condition that the surface enhanced Raman scattering signal is detected at the wavelength of the light source for detection can be found. have.

Metal particles such as silver and gold may be used as the nanoparticles, and the surface of the nanoparticles is coated with a Raman probe for outputting a specific Raman scattering signal, so that the Raman scattering signal is output. As the Raman probe molecule outputting the Raman scattering signal, R6G (rhodamine-6-G), FITC (fluorescein isothiocyanate), MGITC (malachite green isothiocyanate), or the like may be used.

Figure 8a is a graph showing the intensity of the Raman scattering signal according to the concentration of the target material. As the concentration of the target substance increases from 0 ng / mL to 1.5 ng / mL, it can be seen that the Raman scattering signal increases. The Raman scattering signal is a Raman scattering signal of the MGITC Raman probe molecule, and a laser of 647 nm wavelength was used as incident light.

FIG. 8B is a graph illustrating the magnitude of a specific peak signal (1615 cm ⁻¹ ) among the Raman scattering signals and comparing the concentration with the concentration of the target material. As the concentration of AFP, the target substance, increases, the magnitude of a specific peak signal of the Raman scattering signal increases linearly. At this time, the detection limit for the target material AFP was about 20 pg / mL it was confirmed that the very excellent performance.

FIG. 9 is a fluorescence image obtained by simultaneously capturing a large number of microparticles and simultaneously washing the probe nanoparticles which do not stick using the photoelectrofluid controlled immunoassay device according to the present invention.

When performing simultaneous multiple detection using single microparticles as described above, more accurate quantitative detection is possible using confocal fluorescence microscopy. When performing simultaneous multiple detection as described above, a method of immobilizing various kinds of antibodies on the surface of the ground electrode layer 120 or the photoconductive layer 110, rather than the antibody-immobilized microparticles as a support layer, may be used.

FIG. 10 is a conceptual diagram illustrating a photoelectrofluid controlled immunoassay method using the antibody 144 immobilized on the surface of the electrode 122 of the ground electrode layer 120.

In the case of the photoelectrofluid controlled immunoassay method using the immobilized antibody, the sample is characterized in that it comprises an analyte 142 and probe microparticles 143 for measurement. In this case, the antibody 144 immobilized on the surface of the electrode and the probe microparticle 143 may form an immunocomplex 140 via the target material.

The antibody 144 may be immobilized on the surface of the electrode 122 of the ground electrode layer 120 or the photoconductive material 113 of the photoconductive layer 110, and various kinds of antibodies may be patterned in an array form. Can be. In the case of using various kinds of antibodies, the light pattern A is formed to correspond to the pattern form of the antibody 144 to simultaneously control the reaction of antigens to each antibody, and to wash and detect the probe microparticles 143. This is possible at the same time.

The sample may also be injected simultaneously with a plurality of samples to detect several kinds of antigens present in each sample at the same time. In this case, a microstructure (not shown) or a superhydrophobic material (not shown) may be formed between the photoconductive layer 110 and the ground electrode layer 120 to separate the plurality of samples. In addition, it may include an injection port (not shown) for injecting each sample, and may include a microfluidic flow path (not shown) for moving each sample.

As described above with reference to the drawings illustrating an immunoassay device and an immunoassay method using the photoelectromagnetic fluid control technology according to the present invention and the embodiment according to this, for the purpose of illustrating the present invention in more detail, the present specification The present invention is not limited to the embodiments and drawings disclosed in, and various modifications may be made by those skilled in the art within the technical scope of the present invention.

Claims (35)

A light source for irradiating light;
A pattern forming apparatus positioned in a path of light irradiated from the light source to change the pattern of the light;
A photovoltaic fluid device in which a light pattern passing through the pattern forming apparatus is irradiated and a sample is injected; And
Optoelectronic fluid control immunoassay device comprising a power supply for applying power to the optoelectronic fluid device. The method of claim 1,
And at least one condensing lens for condensing the light pattern output from the pattern forming apparatus between the pattern forming apparatus and the optoelectronic fluid element. The method of claim 2,
And a stage in which the optoelectronic fluid element is positioned, wherein the stage is movable up, down, left, and right to change a light pattern region irradiated through the condensing lens. The method of claim 1,
The pattern forming apparatus includes a device driver that can be adjusted by a computer, thereby enabling automated image manipulation. The method of claim 1,
And a detection light source for irradiating incident light to the photoelectromagnetic fluid element to optically observe the sample. The method of claim 5, wherein
And a signal detector for detecting a signal due to a material electronic state or a material vibration state generated by the incident light. The method according to claim 6,
The material electronic state is fluorescence emission or light absorption by wavelength, and the material vibration state is Raman scattering or IR absorption. The method according to claim 6,
The signal detector is any one selected from CCD, photodiode, photomultiplier tube and spectrometer. The method according to claim 6,
And the signal detector comprises a signal processor, the signal processor processing and storing the received signal to generate data. The method of claim 5, wherein
And at least one focusing lens for focusing the incident light irradiated from the detection light source and the output signal output from the photoelectric fluid element. The method of claim 5, wherein
And said focusing lens for focusing said incident light and said output signal is a plurality of different lenses. The method of claim 5, wherein
And a first mirror for reflecting the incident light so that the incident light output from the detection light source is irradiated on the photoelectric fluid element and the sample. The method according to claim 6,
And a second mirror for reflecting the output signal output from the sample to the signal detector. The method of claim 5, wherein
The detection light source further comprises a shutter for adjusting the irradiation time of the incident light, the shutter is opened after the voltage is cut off to the optoelectronic fluid device, characterized in that the opto-fluid control immunoassay device. The method according to claim 6,
The signal detector further includes a shutter for adjusting a reception time of the output signal, the shutter is opened after the voltage is cut off to the optoelectronic fluid element. The method according to claim 6,
The signal detector further comprises a filter or a pinhole for filtering the output signal output from the sample. The method according to claim 14 or 15,
And the light source, the detection light source, the signal detector, the stage, and the shutter are synchronized by the computer-controlled control module and automatically adjusted. The method of claim 1,
The optoelectronic fluid device,
A photoconductive layer positioned on a surface to which the light pattern is irradiated and having a current conducting only to an area to which the light pattern is irradiated;
A ground electrode layer disposed on a surface opposite the photoconductive layer and forming an electric field in the sample from a voltage applied to the photoconductive layer; And
And a space layer formed between the photoconductive layer and the ground electrode layer and spaced apart from the photoconductive layer and the ground electrode layer. The method of claim 18,
The photoconductive layer,
And a photoconductive material for selectively applying a voltage only to a region to which the light pattern is irradiated, the plate electrode applying a voltage by using the power supply device. The method of claim 19,
And the photoconductive material is any one selected from hydrogenated intrinsic amorphous silicon, cadmium sulfide, and npn phototransistor. The method of claim 18,
The photoconductive layer further comprises a doped intermediate layer for reducing the contact resistance between the photoconductive material and the electrode between the photoconductive material and the plate electrode. The method of claim 21,
The intermediate layer is an opto-fluid controlled immunoassay device, characterized in that consisting of amorphous silicon or molybdenum. The method of claim 18,
And a protective layer for protecting the photoconductive material between the photoconductive layer and the space layer, wherein the protective layer is made of silicon nitride or silicon oxide. The method of claim 18,
The space layer further comprises a microstructure or a superhydrophobic material for spaced apart a plurality of samples in the space layer. The method of claim 1,
The sample is a photoelectron fluid controlled immunoassay device, characterized in that it comprises a microparticles, analyte and the probe probe microparticles to be a support layer. The method of claim 25,
The amount of the sample is characterized in that the 100nl ~ 1000nl, the concentration of the analyte can be detected even in the case of ultra-low microscopically low to the level of Atomol (aM). The method of claim 25,
The microparticles serving as the support layer and the probe microparticles form an immunocomplex through the analyte. The method of claim 25,
The microparticles are photoelectron fluid controlled immunoassay device, characterized in that the polymer microparticles are polystyrene or latex. The method of claim 25,
The probe microparticles are fluorescent nanoparticles or Raman scattering material labeled metal nanoparticles, characterized in that the photoelectrofluid control immunoassay device. The method of claim 25,
The substance to be analyzed is an opto-fluid controlled immunoassay device, characterized in that any one or more of an antigen-antibody, DNA, RNA and protein. The method of claim 18,
In the space layer, when the voltage is applied to the space layer by the power supply device and the pattern forming device and the light pattern is irradiated, an electric field is formed from a sample injected into the space layer, and an electrodynamic phenomenon occurs. Immunoassay device. Injecting samples into the optoelectronic fluid device;
Irradiating a light pattern on the optoelectronic fluid device;
Applying a voltage to the optoelectronic fluid device;
Generating an electric field in the optoelectronic fluid device by the voltage and the irradiated light pattern, and causing the samples in the optoelectronic fluid device to flow to induce an antigen-antibody reaction;
Moving the region irradiated with the light pattern to concentrate the antigen-antibody reacted samples into a region irradiated with the light pattern;
Washing the probe microparticles and the target materials which did not react in the concentration step by moving in a direction in which the light pattern is irradiated or in the opposite direction;
Irradiating detection incident light on the sample to detect the amount of the probe microparticles bound to the microparticles or the support in the concentration step; And
And outputting a signal due to photo-material interaction between the incident light and the sample, and detecting the output signal by a detector. 33. The method of claim 32,
The light pattern irradiation step is a photoelectron fluid control immunoassay method characterized in that for irradiating a light pattern on a portion of the optoelectronic fluid device. The method of claim 32,
The reaction step and the concentration step apply a frequency voltage in the range of 1 kHz to 1 MHz, and the washing step is due to the frequency voltage in the range of 100 Hz to 1 kHz, so that the sample is subjected to electrophoresis or electrolysis within the applied frequency range, respectively. Optoelectronic fluid-controlled immunoassay, characterized in that to produce osmotic flow. 33. The method of claim 32,
The washing step is further applied a frequency voltage in the range of 10 kHz ~ 100 kHz, the photoelectron fluid control immunoassay, characterized in that to reassemble or re-concentrate the fine particles within the frequency range.

Images