TWM488629U - Sensing device - Google Patents

Abstract

A sensing device for identifying quantity and quality of a target object, wherein the target object is selected from the group consisting of a protein, a cell, a compound, a metal ion, and a combination thereof. The sensor device includes a detachable chip, the detachable chip includes a substrate, and a nano-particle unit, wherein the substrate is made of a transparent material, and the nano-particle unit is arranged on the substrate and includes a plurality of separated nanometers; an element with a hole, the detachable chip detachably arranged on one end of the element with a hole to form a complex element; and a framework, wherein the complex element is disposed in the framework, and the sensor device is placed in an outer spectrometer to read a value.

Description

Sensing device

The present invention relates to a sensing device, and more particularly to a sensing device that utilizes local surface plasma resonance.

At present, experimental orifice plates on the market have various structures and materials due to different experimental requirements. For example, there are 6, 12, 24, 48, 96, 384 and 1536 holes in the number of holes; The bottom structure is divided into a flat bottom (Flat bottom), a round bottom (Vounded bottom), a V-bottom (V-bottom), and an easy-to-clean bottom combined with a round bottom and a flat bottom; the polystyrene is divided by a material. , PS), polypropylene (PP), polyvinyl chloride (polyvinyl chloride), etc.; color, transparent, black, white, black transparent bottom and white transparent bottom; There are general analysis, cell culture and cell analysis, immunoassay and preservation. Immunoassays generally well plate with most of polystyrene material, for the structure of a multi-well plate 96, but another called Stripwell ^TM (Corning) of the structure, consisting of a strip-shaped object 8 having a hole of a shaped article and The bracket is composed of the strip member and can be attached to the strip member bracket as needed. Generally, the bottom surface of the well plate for immunoassay is un-treated or irradiated to produce a carboxyl group and a hydroxyl group on the benzene ring on the surface of the original well plate. It has an increased ability to bind molecules to which it is to be coated.

Enzyme-linked immunosorbent assay (ELISA) is a common sensing method. It has been used for many years. It includes at least two methods: antigen or antibody, which are discussed as follows:

1. When the sample to be tested is an antigen, the enzyme-linked immunosorbent method comprises the following steps: (1) coating the specific antibody on the plastic orifice plate, and the fixation time is about 12-18 hours. After the fixation is completed, the excess antibody is washed away; (2) the test substance and the immobilized antibody are added to carry out the reaction, and the reaction time is about 0.5-2 hours, and the antigen to be reacted with the immobilized antibody is reactive. , it will be specifically bonded with the antibody immobilized on the plastic well plate; (3) wash away the excess analyte, and add an antibody with an enzyme reactive with the antigen to bond with the antigen, bonding The time is about 0.5-1 hour; (4) Wash away the excess unbound antibody with enzyme, add the enzyme substrate to make the enzyme color, and the coloring time takes about 0.5 hours to read the coloring result by spectrometer (ie The absorbance value (OD value)), the experiment is completed in about 1-2 days in total.

2. When the sample to be tested is an antibody, the enzyme-linked immunosorbent method comprises the following steps: (1) coating the known antigen on the plastic orifice plate, and the fixing time takes about 12-18 hours to complete. After washing off the excess antigen; (2) adding the test substance and the immobilized antibody for reaction, the reaction time is about 0.5-2 hours, and if the sample contains a reactive primary antibody, the antibody It will be specifically bonded to the antigen immobilized on the plastic orifice plate; (3) Wash off the excess analyte, add the secondary antibody with enzyme, and bond with the primary antibody to be tested, the bonding time is about 0.5-2 hours; (4) wash away the excess unbonded twice. The antibody is added to the enzyme to make the enzyme color, and the coloring time takes about 0.5 hours. The coloring result (ie, the absorbance value (OD value)) is read by a spectrometer, and the experiment is completed in about 1-2 days.

The user of the enzyme-linked immunosorbent assay is the aforementioned orifice plate for immunoassay, whether the well plate is unmodified or irradiated, and the antibody or antigen is initially coated in the well. The steps of the disk are combined by physical adsorption. This physical adsorption is non-specific, so it takes up to 12-18 hours of reaction time, and the reaction time of the antibody with the enzyme and the enzyme substrate is also included. This allows the entire experiment to be completed for up to 1-2 days, and requires the use of enzymes and enzymes that are not expensive, so enzyme-linked immunosorbent assays have room for improvement in both time and price.

Surface plasmon resonance (SPR) is a sensing technology developed in recent years. The principle is that when an external light source is irradiated onto a metal film having a nanostructure at any angle, such as a wavelength and a metal surface. When the free electron resonance wavelength is the same, the free electrons are excited to generate collective oscillation and cause the absorption of light to generate the wavelength λ1. Once the metal surface is bonded to the biological or chemical molecule, the wavelength λ1 is shifted to λ2, by detecting The change in wavelength is known about the nature and concentration of the analyte. The time required for surface plasma resonance is shorter than the enzyme-linked immunosorbent method, and the surface plasma resonance needs to be performed by a dedicated instrument, so the price is relatively high, and the implementation is also inconvenient.

After surface plasmon resonance (SPR), localized surface plasmon resonance (LSPR) has been developed, and local surface plasma resonance has many Advantage. The principle is that when the metal nanoparticle is fabricated on a transparent substrate, the excitation of the incident light will cause surface plasma resonance on the surface of the nanoparticle, and the frequency and intensity of the resonance are easily affected by the surrounding environment to generate a wavelength shift or The change in signal intensity, etc., so that the change in local dielectric constant can be used to detect the analyte. Optical changes can be measured by the optics as long as the analyte is bonded to the particle. The surface of the nanoparticle is like a tiny detector that can measure very high optical changes within a few nanometers.

The main difference between local surface plasma resonance (LSPR) and surface plasma resonance (SPR) is that the distance from the surface plasma can be detected differently. The surface plasma penetration (SPR) plasma field penetration depth is between 200- Between 1000nm, the local surface plasma resonance (LSPR) is only between 15-30nm, so the effect of local surface plasma resonance (LSPR) on the far surface is far less sensitive than surface plasma resonance (SPR), in other words Local Surface Plasma Resonance (LSPR) detects only changes in proximity to the surface, thus allowing for complex or impure reaction solutions.

Table 1 compares the current three sensing mechanisms for intermolecular interaction identification, including enzyme-linked immunosorbent assay (ELISA), surface plasma resonance (SPR), and local surface plasma resonance (LSPR). Surface Plasma Resonance (LSPR) performed very well in every project: Partial Surface Plasma Resonance (LSPR) is calibration-free and can be monitored in real time compared to enzyme-linked immunosorbent assay (ELISA). Surface plasma resonance (SPR) is not required for temperature control, and the cost of local surface plasma resonance (LSPR) is also lower than that of enzyme-linked immunosorbent assay (ELISA) and surface plasma resonance (SPR). However, there are still many problems to be solved in commercializing local surface plasma resonance (LSPR).

Table 1 Enzyme linked immunosorbent assay (ELISA), surface plasma resonance (SPR) and local surface plasma

At present, the local surface plasma resonance (LSPR) product on the market is only LamdaGen. The principle is as follows: 1. Provide a three-dimensional structure of the substrate surface, such as undulation wrinkles, micro-aperture, nanowire, etc.; Particles such as gold and silver are adsorbed on the surface of the substrate of the three-dimensional structure as the LSPR sensing material; 3. The surface of the nano metal particles adsorbed on the substrate of the three-dimensional structure is modified to selectively detect molecules, such as DNA, IgG, etc.; 4. Use optical fiber to emit incident light on the nanostructure substrate, and collect the secondary reflection source again, and the displacement between the incident light and the incident light by the spectrometer is used for dynamic monitoring and quantitative analysis. concentration. LamdaGen's products are only It can be read using the company's spectrometer, which is expensive and imposes a heavy burden on the user.

Later, LamdaGen Company also proposed the Optical Enhancement System, which additionally performs antigen-antibody action on the previous measurement antigen step, thus increasing the displacement. However, the Optical Enhancement System can only be read using the company's spectrometer, and the price issue has not been resolved.

Therefore, there are still many problems in the sensing devices currently available on the market, such as time and price to be solved. Although local surface plasma resonance (LSPR) has been introduced, it is not easy to popularize because of its high price and inconvenient use. In order to popularize the local surface plasma resonance (LSPR), the price and ease of use are urgent problems to be solved.

The purpose of the present invention is to provide a sensing device for qualitative and quantitative determination of a test object, wherein the test object is selected from the group consisting of a protein, a cell, a compound, a metal ion, and a combination thereof. The sensing device comprises: a removable wafer, the removable wafer comprises a substrate, and a nano particle unit, wherein the substrate is made of a light transmissive material, and the nano particle unit is disposed on a plurality of spaced apart nanoparticles on the substrate; a porous element detachably disposed at one end of the apertured member to form a composite component; and a frame, wherein the composite The components are assembled into the frame and a value is read by an external spectrometer.

Another object of the present invention is to provide a sensing device comprising: a removable wafer, the removable wafer comprising a substrate, and a nano particle unit, wherein the substrate is made of a light transmissive material. And the nano particle unit is disposed on the substrate and includes a phased complex a plurality of nanoparticles; a perforated component, wherein the removable wafer is removably disposed at one end of the apertured component to form a composite component; and a frame, wherein the composite component is assembled to the frame for sensing.

A further object of the present invention is a sensing device comprising: a removable wafer comprising a nanoparticle unit; and a perforated component, wherein the removable wafer is removably disposed on the apertured component One end is formed to form a composite component for sensing.

A further object of the present invention is a sensing wafer carrier comprising: a perforated component for carrying a removable wafer at one end thereof; and a wafer receiving portion disposed on the apertured component for receiving The detachable chip is disposed on the accommodating portion and is disposed on the accommodating portion for allowing a detecting light when sensing the detachable chip The apertured component and the removable wafer are penetrated.

11‧‧‧Loadable wafers

111‧‧‧Ginnel particles

112‧‧‧Substrate

113‧‧‧antibody

114‧‧‧ antigen

115‧‧‧Analog-labeled gold nanoparticles

12, 22, 52‧‧‧ holed components

121, 221‧ ‧ holes

122, 222, 623‧‧‧Inlay holes

123, 223‧‧‧ grooves

13, 23, 53‧‧‧ composite components

3‧‧‧Frame

31‧‧‧Inlay column

62‧‧‧ Vehicle body

621‧‧‧Whip Capacity Department

622‧‧‧Detecting light penetration

The first figure (a) is a removable wafer of the embodiment of the present creation

The first figure (b) is a perforated component of the embodiment of the present creation

The first figure (c) is a composite component of the embodiment of the present creation

The second figure (a) is a removable wafer of the embodiment of the present invention.

The second figure (b) is a perforated component of the embodiment of the present creation

The second figure (c) is a composite component of the embodiment of the present creation

The third picture is the framework of the embodiment of the present creation

The fourth figure (a) is a schematic diagram of the use of six sets of composite elements 23 when the number of samples is 48 in the embodiment of the present creation.

The fourth figure (b) is a schematic diagram of the use of 12 sets of composite elements 23 when the number of samples is 96 in the embodiment of the present creation.

Figure 5 (a) is a removable wafer of another embodiment of the present invention

Figure 5 (b) is a perforated component of another embodiment of the present invention

Figure 5 (c) is a composite component of another embodiment of the present creation

Figure 6 (a) is a removable wafer of another embodiment of the present invention

Figure 6 (b) is a sensing wafer carrier of another embodiment of the present creation

Figure 6 (c) is a sensing wafer carrier of another embodiment of the present creation

The seventh figure is the substrate coating characteristics of the microwave plasma nanoparticles of the experimental example of the present invention.

The eighth figure is the fabrication and reproducibility of the sensing wafer of the experimental example of the present creation.

The ninth figure (a) is a structural stability test of the sensing wafer of the experimental example of the present creation.

The ninth figure (b) is the surface oxidation effect test of the sensing wafer of the experimental example of the present creation.

The tenth figure is a method for further signal amplification of the sensing chip of the experimental example of the present creation

The technical content, features, and effects of the present invention will be apparent from the following detailed description of the preferred embodiments.

A preferred embodiment of the present invention is a sensing device for qualitative and quantitative determination of a test object, wherein the test object is selected from the group consisting of a protein, a cell, a compound, a metal ion, and a combination thereof. Group of.

As shown in the first (a) to third figures, the sensing device comprises a removable wafer 11, a perforated component 12, 22 (see first (b) and second (b)); A frame 3 (see the third figure). The area of the detachable wafer 11 is 1 to 49 mm ² , and may be, for example, (1 to 7 mm)* (1 to 7 mm). The removable wafer 11 is selected from the group consisting of a circle, an ellipse, a polygon, an irregular shape, and combinations thereof. The detachable wafer 11 comprises a substrate, a nano particle unit and a sensing unit, wherein the substrate is made of a transparent material, and the transparent material of the substrate is selected from polyethylene (polyethylene). , PE), High-density polyethylene, Low-density polyethylene, Polypropylene (PP), Polystyrene (PS), Polyvinyl chloride (poly(vinyl) Chloride), PVC), Polyethylene terephthalate (PET), poly(dimethylsiloxane (PDMS), Polymethylmethacrylate (PMMA), Polycarbonate a group of polycarbonates (PC), glass, quartz, quartz glass, mica, sapphire, transparent ceramics, and combinations thereof, and the nanoparticle unit is disposed on the substrate And comprising a plurality of spaced apart nanoparticles, the method of which can be referred to the patent of the Republic of China No. I404930. Each of the nanoparticles is made of a metal selected from the group consisting of gold and silver. , copper, palladium, platinum, titanium, chromium a group consisting of nickel, zinc, alloys of the above metals, and combinations thereof. One of the nano particles is selected from the group consisting of a circle, an island, a strip, a triangle, a star, a ring, a hollow, and combinations thereof. The group of the nanoparticles has a particle diameter of 1 to 200 nm, and the nanoparticles have a pitch between 1 nm and 100 nm.

The detachable chip 11 includes the sensing unit including a plurality of receivers disposed between the nano particles; the method of the receiver can be referred to the patent of the Republic of China No. I404930. Determining, according to the type of the object to be tested, the receiver coupled to the surface of the substrate, and then transmitting the characteristics of the metal nanoparticles, when the receivers of the sensing unit form a specificity with the object to be tested When combined, the local electromagnetic fields induced by the light of the metal nanoparticles are affected by the light. The environmental influence changes and causes the spectral signal to change. Therefore, the change of the spectral signals of the metal nanoparticles before and after the combination of the receiver and the test object can be used to detect whether the sample contains the analyte and further quantify the sample. Its concentration makes the sensing unit both qualitative and quantitative. For example, when the analyte is streptavidin, the specificity of binding of avidin to biotin can be utilized, and biotin is used as a receiver because biotin cannot form a stable structure directly with the substrate. In combination, 3-aminopropyltriethoxysilane (APTMS), which is more easily bonded to the substrate and bonded to biotin, can be used to form an APTMS molecular film on the surface of the substrate. By adding biotin, biotin can be indirectly modified on the surface of the substrate through APTMS, and the combination of APTMS and biotin is a receiver. In addition, when the analyte is a mercury ion, the specific combination of the mercury ion and the 4-carboxybenzo-15-crown-5 can be utilized. The modification of the sub-mercury ion is also possible by modifying the saline molecule on the surface of the substrate and then attaching 4-carbonate benzo-15-crown-5. At this time, the receivers are a combination of decane modified on the surface of the substrate 2 and 4-carbonic acid benzo-15-crown-5 bonded to the decane. The above-mentioned method of modifying the substrate by appropriate molecular chemical modification can shorten the time for coating the receiver on the substrate to only one hour, and attaching the antibody or antibody to the enzyme-linked immunosorbent method. Compared with 12-18 hours, it takes a significant improvement.

The apertured component has a hole 121, 221 and an engagement hole 122, 222. The number of holes of the apertured component 12, 22 is an integer ranging from 1 to 384. The removable wafer 11 is detachably disposed on the hole. One end of the apertured member 12, 22 is formed to form a composite component 13, 23, and one end of the apertured component 12, 22 can have a recess 123, 223, and the removable wafer 11 can be disposed in the recess 123. , 223 to form the composite component 13, 23, the removable wafer 11 and the apertured component 12, 22 The connection manner may be adhesion, riveting, screwing, welding, inlay and hinge, but is not limited thereto. The composite component 13 and 23 are used to hold the object to be tested. The object to be tested can be directly in contact with the surface of the detachable wafer 11 in the composite component 13 and 23, and the sensing unit of the detachable wafer 11 is known. Whether the receivers can form a specific combination with the object to be tested.

The composite elements 13, 23 are assembled to the frame 3 and a numerical reading is performed by an external spectrometer. Assembly can be by bonding, riveting, screwing, welding, inlaying, and articulating, but is not limited thereto. The assembly method used in this embodiment is a fitting, and the frame 3 has an engaging post 31 and the engaging holes 122, 222 of the perforated member are used in combination. The number of holes and the number of the holes of the porous elements 12, 22 of the composite component 13, 23 can be determined according to the needs of the user, as shown in the fourth figure (a) and the fourth figure (b), when the number of samples For 48 o'clock, 6 sets of the composite element 23 having the number of holes of the perforated element 22 can be used; when the number of samples is 96, 12 sets of the perforated element 22 can be used. Element 23, unlike conventional 96-well plates, requires a full 96-well plate at a time, regardless of the number of samples. When the number of samples is large but the required amount is small or expensive, a composite element having the number of holes of 384 of the perforated member can be used, so that a large number of samples can be processed at one time, and the sample usage can be saved. The value is a wavelength, and the wavelength range read is 300 to 700 nm. The wavelength range to be read varies depending on the particle diameter of the metal nanoparticle (or the thickness of the metal layer) and the material of the metal nanoparticle, for example, when the average particle size of the metal nanoparticle When the diameter is 5 nm to 20 nm, the wavelength range to be read falls within the range of 400 nm to 650 nm. When the total thickness of the metal layer is controlled at 3 nm, the wavelength of the formed gold nanoparticles mainly falls within the range of 510 nm to 540 nm; the wavelength of the nano particles of the formed gold-silver alloy mainly falls within the range of 410 nm to 490 nm. The sensing device of the embodiment has the same length and width as the experimental orifice plate on the market, and thus can be used for any instrument used in conjunction with the experimental orifice plate, such as a spectrometer and an automatic microplate washer, the spectrometer Can be An enzyme linked immunosorbent assay (ELISA reader).

As shown in FIGS. 5(a) to 5(c), a sensing device according to another preferred embodiment of the present invention includes a removable wafer 11 including a nano particle. And a perforated element 52, wherein the removable wafer 11 is sensed by being detachably disposed at one end of the perforated element 52 to form a composite element 53. The number of holes of the apertured component 52 is an integer ranging from 1 to 384. The removable wafer 11 can be set to be between 1 and 384 according to the user's requirements. The sensing device of the embodiment can be used for any instrument used in conjunction with the experimental orifice plate without a frame, such as a spectrometer and an automatic microplate washer, which can be an enzyme-linked immunosorbent reader ( ELISA reader).

As shown in FIG. 6(a) to FIG. 6(c), a sensing wafer carrier according to still another preferred embodiment of the present invention includes: a carrier body 62 for carrying a carrier thereon The detachable wafer 11 is disposed on the carrier body 62 for receiving the detachable wafer 11 and a detecting light transmitting portion 622 disposed on the carrier body 62. When sensing the detachable wafer 11 for sensing, a detection light is allowed to penetrate the carrier body 62 and the detachable wafer 11. The detecting light penetrating portion 622 is penetrated through a hollow portion of the carrier body 62. The carrier body 62 can be a holed component. The carrier body 62 can carry the detachable chip 11 at one end thereof. The detecting light transmitting portion 622 can be located above the chip accommodating portion.

Compared with the enzyme-linked immunosorbent assay (ELISA), the sensing device of the present invention has a secondary antibody and a coloring agent that do not need to be linked to the antigen-measuring antigen, and takes much longer than the enzyme-linked immunosorbent method ( ELISA) has fewer advantages; the sensing device required by the present sensing device is also much less than the enzyme-linked immunosorbent assay (ELISA), and the volume of the analyte is only 20 μL; and it is commercialized now. Compared with the local surface plasma resonance (LSPR) technology, the sensing device of this creation It can be used in a standard enzyme-linked immunosorbent assay (ELISA) system equipped with a general immunoassay laboratory. It does not require the purchase of an expensive proprietary spectral reader, and the sensing device can be designed to operate up to 384 samples at a time. The effect of High throughput screening (HTS) is an absolute advantage in terms of time and price, as well as ease of use.

The sensing device of the present invention has the advantages of less steps, no labeling, low cost, short time consumption, no need to add coloring enzymes, and can be applied to detect different antibodies and viruses.

This creation can be applied to experimental development, such as immunoanalytical chemical analysis and enzyme analysis; establish experimental procedures such as kinetic function and temperature control; antibody identification, such as antibody/ligand affinity screening, monoclonal antibody antigen Epitope determination, tumor cell screening and identification period, anti-unique antibody screening, antibody concentration determination and fragment screening; preclinical and clinical diagnosis, such as biomarker analysis and key care.

Experimental example:

1. Production of Microwave Plasma Nanoparticles: According to the method of the Republic of China No. I404930. First, a gold film is sputtered on the glass substrate, and then placed in a microwave plasma for treatment. The treatment time is only 30 seconds. Under the instant heating state of both microwave and microwave plasma, gold is formed on the glass substrate. The nano particles and the bottom of the gold nanoparticles are coated with a glass structure, which greatly improves the adhesion between the gold nanoparticles and the substrate, which is a characteristic not used in the conventional heating method. It can be seen from the experimental examples that the microwave plasma heating method has the following advantages: low cost, simple and rapid, semi-embedded nano-particles, thus good adhesion and controllable size.

2. Detecting the substrate coating characteristics of microwave plasma nanoparticles: Referring to the seventh figure, first use atomic force microscopy (atomic force) Microscope, AFM) The surface of the gold nanoparticle 111 formed in the experimental example was observed, and it was found that the structure of the gold nanoparticle 111 belongs to an island structure. The substrate 112 is then bubbled into the aqueous solution of the king to remove the gold nanoparticles 111 on the substrate 112. The substrate 112 treated with the aqueous solution of the aqua regia was again observed by atomic force microscopy, and it was found that many annular structures remained on the surface of the substrate 112. These annular structures were made of glass, which indicated that the bottom of the gold nanoparticle 111 was covered with a layer of glass. . This is mainly because when the nanoparticles are treated under microwave and microwave plasma, they will reach a high temperature state instantaneously, just like a nano-droplet that exhibits a high temperature state, which can partially melt the glass substrate by gravity. And the capillary action, the molten glass gradually covers the surface of the nanoparticle, and thus in this experimental example, the island-like structure in which the gold nanoparticle 111 is semi-embedded on the substrate 112 is formed.

3. Fabrication and Reproducibility Detection of Sensing Wafer: Referring to the eighth drawing, in the method of Experimental Example 1, a total of 20 substrates were prepared, the film thickness of which was controlled at 2 nm, and the processing time was 30 seconds. After the 20 substrates were obtained, the optical absorption wavelengths were individually recorded using the spectrum, and as a result, the optical absorption wavelengths of the 20 substrates all fell within the range of 519 ± 1.7 nm, which is a very small distribution range. From this optical distribution, the reproducibility of these 20 substrates is very high, and it has an extremely important potential for developing disposable biosensing wafers.

4. Sensing the structural stability and surface oxidation effect of the wafer: Referring to the ninth figure, the structural stability and surface oxidation effect of the substrate are further identified by the spectrum, firstly the stability of the structure: the sensing chip is most afraid The problem is that the bubble in the solution system will accidentally fall off or the structure is deformed, thus causing errors in the interpretation of the optical signal. In this experimental example, the substrate is washed with ultrapure water, PBS buffer solution and ethanol solution, which does not cause optical significance. The change, the nano structure of the creation is very stable. Due to the high surface activity of the gold nanoparticles, the oxidation effect of the surface was also observed. It was found that the gold nanoparticles were initially in the table. Oxidation of a film on the surface causes a slight change in optics and tends to stabilize after one day, no longer oxidizing. The substrate representing the creation can be placed in the environment for a long time.

5. Modifying the created sensing wafer with 3-aminopropyltriethoxydecane for the detection of mercury ions: firstly, the surface of the substrate of the sensing wafer of the present invention is made hydrophilic by the weaker oxygen plasma. Sexually modified, and then the substrate is bubbled into 3-aminopropyltrimethoxysilane (APTMS) solution to make 3-aminopropyltriethoxysilane and nanoparticle The blank substrate is bonded, and then 4-carbon benzo-15-crown-5 is attached to form a receiver, and finally, the mercury ion is added to react with the receiver to perform sub- Mercury ion detection experiment.

6. Modifying the created sensing wafer with 3-aminopropyltriethoxydecane for the detection of avidin: firstly, the surface of the substrate of the sensing wafer of the present invention is made hydrophilic by the weaker oxygen plasma. After upgrading, the substrate can be bubbled into 3-aminopropyltrimethoxysilane (APTMS) solution to make a gap between 3-aminopropyltriethoxysilane and nanoparticle. The substrate is bound, and then NHS-biotin (N-hydroxy-succinimide-biotin) is added to form a receptor with APTMS to form a receiver, and finally Streptavidin is added to react with the receiver to perform detection of avidin. Test experiment.

7. Modifying the created sensing wafer with 3-aminopropyltriethoxydecane for antigen or antibody detection experiments: firstly, the surface of the substrate of the sensing wafer of the present invention is made hydrophilic with a weaker oxygen plasma. Sexually modified, then inoculate the substrate into 3-aminopropyltriethoxydecane In a (3-aminopropyl)trimethoxysilane (APTMS) solution, a 3-aminopropyltriethoxysilane can be bonded to a blank substrate between the nanoparticles, followed by the addition of glutaraldehyde (GA) and APTMS. An imine linkage, followed by addition of an antibody (antigen) to form a receiver, and finally an antigen (antibody) to be tested is reacted with the receiver for antigen or antibody detection experiments.

8. Further signal amplification of the sensing wafer: see the tenth figure, which is different from the traditional modification of the antibody molecule on the surface of the gold nanoparticle 111. The sensing device of the present invention will be 3-aminopropyltriethoxydecane (3). -aminopropyl)trimethoxysilane, APTMS) is modified on the interstitial substrate of the nanoparticle, and uses Glutaraldehyde (GA) as a linker molecule linking APTMS to the antibody to link the antibody 113, and the antigen 114 can be carried out. Capture, after the antigen 114 is replenished, in order to further observe a smaller amount of the analyte, the antibody-labeled gold nanoparticle 115 is again added to form a sandwich sandwich structure, due to the antibody-labeled gold nanoparticle 115 and the bottom of the substrate. The semi-embedded gold nanoparticles 111 generate surface-plasma coupling resonance effects with each other, resulting in extremely high optical variations. By performing spectral measurements, the signal is amplified by a thousand times and the sensitivity reaches picomole. The level. Glutaraldehyde (GA) can also be used as a linker molecule that binds APTMS to an antigen to bind an antigen, and even after antibody capture, after further antibody capture, in order to further observe a smaller amount of the analyte, it is added again. Antigen-labeled gold nanoparticles, in this case can also amplify the signal by a thousand times, the sensitivity reaches the level of pico.

Example:

A sensing device for qualitative and quantitative determination of a test object, wherein the test object is selected from the group consisting of a protein, a cell, a compound, a metal ion, and a combination thereof. The sensing device comprises: a removable wafer, the removable wafer comprises a substrate, and a nano particle unit, wherein the substrate is made of a transparent material, and the nano particle is The unit is disposed on the substrate and includes a plurality of spaced apart nanoparticles; a porous member detachably disposed at one end of the apertured member to form a composite member; and a frame Wherein the composite component is assembled to the frame and a value is read by an external spectrometer.

2. The sensing device of embodiment 1, wherein the removable wafer further comprises a sensing unit, the sensing unit comprising a plurality of receivers disposed between the plurality of nano particles.

3. The sensing device according to embodiment 1, wherein the size of the removable wafer is (1 to 7 mm)* (1 to 7 mm).

4. The sensing device of embodiment 1, wherein the removable wafer is selected from the group consisting of a circle, an ellipse, a polygon, an irregular shape, and combinations thereof.

5. The sensing device according to embodiment 1, wherein each of the nanoparticles is made of a metal selected from the group consisting of gold, silver, copper, palladium, platinum, titanium, chromium, nickel, zinc, and the like. A group of alloys of metals and combinations thereof.

6. The sensing device of embodiment 1, wherein the number of holes of the apertured element is an integer between 1 and 384.

7. The sensing device of embodiment 1, for use in a spectrometer and an automatic microplate washer.

8. A sensing device comprising: a detachable wafer comprising a substrate and a nano particle unit, wherein the substrate is made of a light transmissive material, and the nano particle unit is disposed on the substrate and comprises a plurality of spaced apart nanoparticles; a perforated component, wherein the removable wafer is removably disposed at one end of the apertured component to form a composite component; and a frame, wherein the composite component is assembled to the frame for performing Sensing.

9. The sensing device of embodiment 8, further comprising a sensing unit, the sensing unit comprising a plurality of receivers disposed between the plurality of nanoparticles.

10. The sensing device according to embodiment 8, wherein the size of the removable wafer is (1 to 7 mm)* (1 to 7 mm).

11. The sensing device of embodiment 8, wherein each of the nanoparticles is made of a metal.

12. The sensing device of embodiment 8, wherein the number of holes of the apertured element is an integer between 1 and 384.

13. The sensing device of embodiment 8 for use in a spectrometer and an automated microplate washer.

14. A sensing device comprising: a removable wafer comprising a nanoparticle unit; and a perforated element, wherein the removable wafer is removably disposed at one end of the apertured member to form a composite The component is used for sensing.

15. The sensing device of embodiment 14, further comprising a frame, wherein the frame is for assembling the composite component, the removable wafer further comprising a substrate, the substrate is transparent The material is made of a material, and the nano particle unit is disposed on the substrate and includes a plurality of spaced nano particles.

16. The sensing device of embodiment 14 for use in a spectrometer and an automated microplate washer.

17. A sensing wafer carrier comprising: a perforated component for carrying a removable wafer at one end thereof; a wafer receiving portion disposed on the apertured component for receiving the removable wafer And a detecting light transmissive portion disposed on the apertured component and located above the wafer housing portion for allowing a detection light to penetrate the aperture when the removable wafer is sensed Components and the removable wafer.

18. The sensing wafer carrier of embodiment 17, wherein the detecting light penetrating portion extends through a hollow portion of the apertured member.

221‧‧‧ hole

222‧‧‧Inlay hole

23‧‧‧Composite components

Claims (18)

A sensing device is used for qualitative and quantitative determination of a test object, wherein the test object is selected from the group consisting of a protein, a cell, a compound, a metal ion, and a combination thereof, and the sensing device comprises a detachable wafer comprising a substrate and a nano particle unit, wherein the substrate is made of a light transmissive material, and the nano particle unit is disposed on the substrate And comprising a plurality of spaced apart nanoparticles; a porous member detachably disposed at one end of the apertured member to form a composite component; and a frame, wherein the composite component is assembled to the frame A value reading is performed by an external spectrometer. The sensing device of claim 1, wherein the removable wafer further comprises a sensing unit, the sensing unit comprising a plurality of receivers disposed between the nano particles. The sensing device according to claim 1, wherein the size of the detachable wafer is (1 to 7 mm)* (1 to 7 mm). The sensing device of claim 1, wherein the removable wafer is selected from the group consisting of a circle, an ellipse, a polygon, an irregular shape, and combinations thereof. The sensing device according to claim 1, wherein each of the nano particles is made of a metal selected from the group consisting of gold, silver, copper, palladium, platinum, titanium, chromium, nickel, zinc, A group consisting of the above alloys of metals and combinations thereof. The sensing device according to claim 1, wherein the number of holes of the apertured component is an integer between 1 and 384. The sensing device according to claim 1 is used for a spectrometer and an automatic microplate washer. A sensing device comprising: a removable wafer, the removable wafer comprising a substrate, and a nano particle unit, wherein the substrate is made of a light transmissive material, and the nano particle unit is disposed on a plurality of nanoparticles spaced apart from the substrate; a porous element, wherein the removable wafer is detachably disposed at one end of the apertured member to form a composite component; and a frame, wherein the composite Components are assembled to the frame for sensing. The sensing device of claim 8, wherein the removable wafer further comprises a sensing unit, the sensing unit comprising a plurality of receivers disposed between the nano particles. The sensing device according to claim 8 is characterized in that the size of the removable wafer is (1 to 7 mm)* (1 to 7 mm). The sensing device according to claim 8, wherein each of the nanoparticles is made of a metal. The sensing device of claim 8, wherein the number of holes of the apertured component is an integer between 1 and 384. The sensing device of claim 8 is for use in a spectrometer and an automatic microplate washer. A sensing device comprising: a removable wafer comprising a nanoparticle unit; and a perforated component, wherein the removable wafer is detachably disposed on the apertured component The ends are formed by forming a composite component. The sensing device of claim 14, further comprising a frame, wherein the frame is used to assemble the composite component, the removable wafer further comprises a substrate, the substrate is made of a light transmissive material And the nanoparticle unit is disposed on the substrate and comprises a plurality of spaced apart nanoparticles. The sensing device of claim 15 is for use in a spectrometer and an automatic microplate washer. A sensing wafer carrier comprising: a perforated component for carrying a removable wafer at one end thereof; a wafer receiving portion disposed on the apertured component for receiving the removable wafer; a detecting light transmissive portion is disposed on the apertured component and located above the wafer housing portion for allowing a detection light to penetrate the apertured component when sensing the removable wafer The removable wafer. The sensing wafer carrier of claim 17, wherein the detecting light transmitting portion penetrates a hollow portion of the apertured member.