TWI585390B - Self-referencing localized plasmon resonance sensing device and system thereof - Google Patents

Description

Self-correcting localized plasma resonance sensing device and system thereof

The invention relates to a localized plasma resonance sensing device and a system thereof; in particular, to a self-correcting type localized plasma resonance sensing device and a system thereof.

The electron cloud on the surface of the metal nanoparticles can be excited by an electromagnetic field of a specific frequency, which is collectively oscillated and resonated with the conductive electrons confined in the nanoparticle volume, so it is also called Localized Plasmon Resonance (Localized Plasmon Resonance, LPR), as shown in Figure 1. The noble metal nanoparticles 1 produce a strong absorption band in the absorption spectrum, which is called a localized plasma resonance band. The basic principle of the localized plasma resonance band sensing system is: the object to be tested is covered by the noble metal nanoparticle 1 through the bonding of the identification element on the surface of the noble metal nanoparticle 1 and the object to be detected combined with the identification element. Near the surface, the surrounding dielectric environment of the noble metal nanoparticles 1 changes, and the peak wavelength position and absorbance are quite sensitive to changes in the external dielectric constant, thus causing a change in the LPR resonance band; The specific identification element has the means of detecting the specificity, and by analyzing the relationship between the change of the frequency or intensity of the resonance band and the concentration of the analyte, a quantitative detection method can be established. The method primarily comprises modifying noble metal nanoparticles onto an optical waveguide to form a layer of noble metal nanoparticles thereon. The noble metal nanoparticle layer is composed of spherical noble metal nanoparticles, square noble metal nanoparticles, pyramidal noble metal nanoparticles, One of the rod-shaped noble metal nanoparticles and the shell noble metal nanoparticles, and the nanoparticles are not substantially connected, and the noble metal may be gold, silver or platinum. By utilizing the effect of multiple total internal reflection along the optical waveguide, the amount of change in evanescent wave absorption due to absorption of the plasma resonance of the nanoparticle can be accumulated to increase the signal for detecting the LPR of the operation. At the same time, through the surface modification of the noble metal nanoparticles 1 having various identification elements, the functionalized noble metal nanoparticles can be used for the detection of various analytes.

Single-fiber LPR sensing systems lack the ability to compensate for the effects of the instrument itself or environmental factors on the signal, such as baseline drift due to instability of the source, and changes in composition or temperature of the test solution, and LPR sensing The technique utilizes the sensitivity of precious metal nanoparticles to the external refractive index of the environment as a means of detecting biomolecules, which also depends on the temperature or composition of the sample. During the detection of a real sample, it is usually necessary to control the temperature of the sample often or via two or more dilutions in the sample preparation process. Adding a temperature control system may increase the complexity of the system, and multiple dilutions tend to degrade the effective detection limits.

In view of the above-mentioned problems of the prior art, it is an object of the present invention to provide a self-correcting type localized plasma resonance sensing device and system thereof for eliminating interference caused by environmental factors or inherent dielectric properties of the sample itself. And solve the problem of non-specific adsorption.

According to one of the objects of the present invention, a self-correcting type localized plasma resonance sensing device is proposed. The self-correcting type localized plasma resonance sensing device includes: a reference optical waveguide element, a sensing optical waveguide element, and a carrier. The optical waveguide element may be an optical fiber, a channel waveguide, a planar waveguide, or a tubular waveguide or the like. Preferably, the optical waveguide component is an optical fiber. The reference optical waveguide element is modified with a first noble metal nanoparticle layer. A portion of the incident light is directed into the reference optical waveguide component to produce a first localized plasma resonant sensing signal, wherein the light is totally internally reflected along the reference optical waveguide component a plurality of times. The first domain plasma resonance sensing signal includes a first signal generated by the reference optical waveguide component detecting the blank solution and a sample generated by the reference optical waveguide component The second signal of birth. The reference optical waveguide element has a first correction slope. Further, the sensing optical waveguide element is modified with a second noble metal nanoparticle layer. The second precious metal nanoparticle layer is further modified with an identification element. Another portion of the incident light is directed into the sensing optical waveguide component to produce a second localized plasma resonant sensing signal, wherein the light is totally internally reflected along the sensing optical waveguide component a plurality of times. The second localized plasma resonance sensing signal includes a third signal generated by the sensing optical waveguide component detecting the blank solution and a fourth signal generated by the sensing optical waveguide component detecting the sample. The sensing optical waveguide component has a second correction slope. The processor normalizes the first difference between the second signal and the first signal and normalizes the second difference between the fourth signal and the third signal. The processor adjusts the normalized second difference using the first corrected slope and the second corrected slope for obtaining the adjusted second difference. The processor then utilizes the difference between the normalized first difference and the adjusted second difference to obtain a sensor response. Further, the carrier places the reference optical waveguide element and the sensing optical waveguide element.

Preferably, the first noble metal nanoparticle layer is modified on the reflective surface of the reference optical waveguide element.

Preferably, the second precious metal nanoparticle layer is modified on the reflective surface of the sensing optical waveguide component.

Preferably, when the reference optical waveguide component is an optical fiber, the first noble metal nanoparticle layer can be modified at the stripping region or end face of the optical fiber.

Preferably, when the sensing optical waveguide component is an optical fiber, the second noble metal nanoparticle layer can be modified at the stripping region or end face of the optical fiber.

Preferably, the reference optical waveguide component and the sensing optical waveguide component are optical fibers, channel waveguides, planar waveguides, or tubular waveguides.

Preferably, the self-correcting type localized plasma resonance sensing device is a microfluidic wafer or a current sampling and analysis device.

Preferably, the reference optical waveguide component and the sensing optical waveguide component respectively construct a mirror surface on one end surface of the reference optical waveguide component and one end surface of the sensing optical waveguide component, wherein the reference optical waveguide component and the sensing optical waveguide component are optical fibers or tubular waveguides.

Preferably, the reference optical waveguide component and the sensing optical waveguide component are further provided with a filter membrane and a rigid bracket having at least one opening. The mirror is provided to reflect the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal, and the filter filters the interfering substance having a size larger than the average pore diameter of the film. In addition, the rigid support encases the reference optical waveguide component and the sense optical waveguide component to increase the mechanical strength of the device during the sampling operation.

Preferably, the identifier comprises a chemical recognition molecule, an antibody, an antigen, a lectin, a hormone receptor, a nucleic acid or a saccharide.

Preferably, the sensor response is represented by the following formula: Where ΔI _{S, SA} represents the sensor response caused by only specific adsorption, I _S0 represents the third signal generated by the sensing optical waveguide component detecting the blank solution, and ΔI _S represents the relationship between the fourth signal and the third signal. The second difference, ΔI _{S, M} /I _S0 and ΔI _{R, M} /I _R0 respectively represent the normalized response of the sensing optical waveguide element and the reference optical waveguide element, which respectively represent the second correction slope and the first correction slope , ΔI _R represents a first difference between the second signal and the first signal, and I _R0 represents a first signal generated by the reference optical waveguide element detecting the blank solution.

According to another object of the present invention, a self-correcting type localized plasma resonance sensing device is proposed. The self-correcting type localized plasma resonance sensing device comprises a light source, a localized plasma resonance sensing device, at least one light detecting unit, and a processor, and the light detecting unit is preferably a photodiode. Light source produces incident Light. The localized plasma resonance sensing device includes a reference optical waveguide component, a sensing optical waveguide component, and a carrier. Here, the reference optical waveguide element is modified with a first noble metal nanoparticle layer. A portion of the incident light is directed into the reference optical waveguide component to produce a first localized plasma resonant sensing signal. The reference optical waveguide element has a first correction slope. Further, the sensing optical waveguide element is modified with a second noble metal nanoparticle layer. The second precious metal nanoparticle layer is further modified with an identification element. Another portion of the incident light is directed into the sensing optical waveguide component to produce a second localized plasma resonant sensing signal. The sensing optical waveguide component has a second correction slope. Further, the carrier places the reference optical waveguide element and the sensing optical waveguide element. In addition, the at least one photo detecting unit receives the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal. Here, the first localized plasma resonance sensing signal includes a first signal generated by the reference optical waveguide component detecting the blank solution and a second signal generated by the reference optical waveguide component detecting the sample. The second localized plasma resonance sensing signal includes a third signal generated by the sensing optical waveguide component detecting the blank solution and a fourth signal generated by the sensing optical waveguide component detecting the sample. In addition, the processor normalizes the first difference between the second signal and the first signal and normalizes the second difference between the fourth signal and the third signal. Here, the processor adjusts the normalized second difference using the first corrected slope and the second corrected slope for obtaining the adjusted second difference. The processor then utilizes the difference between the normalized first difference and the adjusted second difference to obtain a sensor response.

Preferably, the first noble metal nanoparticle layer is modified on the reflective surface of the reference optical waveguide element.

Preferably, the second precious metal nanoparticle layer is modified on the reflective surface of the sensing optical waveguide component.

Preferably, when the reference optical waveguide component is an optical fiber, the first noble metal nanoparticle layer can be modified at the cladding stripping region or end face of the optical fiber.

Preferably, when the sensing optical waveguide component is an optical fiber, the second noble metal nanoparticle layer can be modified at the cladding stripping region or end surface of the optical fiber.

Preferably, the reference optical waveguide component and the sensing optical waveguide component are optical fibers, channel waveguides, planar waveguides, or tubular waveguides.

Preferably, the self-correcting type localized plasma resonance sensing device is a microfluidic wafer or a current sampling and analysis device.

Preferably, the reference optical waveguide component and the sensing optical waveguide component respectively construct a mirror surface on one end surface of the reference optical waveguide component and one end surface of the sensing optical waveguide component, wherein the reference optical waveguide component and the sensing optical waveguide component are optical fibers or tubular waveguides.

Preferably, the reference optical waveguide component and the sensing optical waveguide component are further provided with a filter membrane and a rigid bracket having at least one opening. The mirror is provided to reflect the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal, and the filter filters the interfering substance having a size larger than the average pore diameter of the film. The rigid support encases the reference optical waveguide component and the sense optical waveguide component to increase the mechanical strength of the device during the sampling operation.

Preferably, the identifier comprises a chemical recognition molecule, an antibody, an antigen, a lectin, a hormone receptor, a nucleic acid or a saccharide.

Preferably, the self-correcting localized plasma resonance sensing device of the present invention further comprises a lock-in amplifier capable of amplifying the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal and suppressing System noise.

Preferably, the sensor response is represented by the following formula: Where ΔI _{S, SA} represents the sensor response caused by only specific adsorption, I _S0 represents the third signal generated by the sensing optical waveguide component detecting the blank solution, and ΔI _S represents the relationship between the fourth signal and the third signal. The second difference, ΔI _{S, M} /I _S0 and ΔI _{R, M} /I _R0 respectively represent the normalized response of the sensing optical waveguide element and the reference optical waveguide element, which respectively represent the second correction slope and the first correction slope , ΔI _R represents a first difference between the second signal and the first signal, and I _R0 represents a first signal generated by the reference optical waveguide element detecting the blank solution.

As described above, the self-correcting type localized plasma resonance sensing device and system thereof according to the present invention may have one or more of the following advantages: (1) The disclosed self-correcting type localized plasma resonance sensing The device and its system can reduce the interference caused by environmental factors or the dielectric properties inherent in the sample itself, and can solve the problem of non-specific adsorption, so that the sensing system has self-correcting ability, thus improving the localized plasma resonance sensing. The device and its system can detect the true sample; and (2) the self-correcting localized plasma resonance sensing device and system thereof can reduce the sample dilution in the sample preparation process when detecting the object to be tested The number of times, thereby improving the detection limit of the sensing operation.

1‧‧‧ precious metal nanoparticles

2, 72‧‧‧ self-correcting fiber-optic localized plasma resonance sensing device

21, 721, 81, 91, 101, 111‧‧‧ reference fiber

211, 221‧‧‧ precious metal nanoparticle layer

22, 722, 82, 92, 102, 112‧‧‧ sensing fiber

2211‧‧‧ Identification element

23, 723‧‧‧ carrier

31‧‧‧Reference optical waveguide components

32‧‧‧Sensing optical waveguide components

33‧‧‧ Carrier

34‧‧‧ channel

35‧‧‧V-shaped channel

37, 36‧‧‧ openings

41‧‧‧sample slot

51‧‧‧Mirror

52, 62‧‧‧ filter

61‧‧‧Rigid bracket

71‧‧‧Light source

73‧‧‧Light detection unit

74‧‧‧Processing unit

75‧‧‧Locking amplifier

76‧‧‧Signal Generator

1 is a schematic diagram of a conventional localized plasma resonance; FIG. 2 is a schematic diagram of a self-correcting fiber-type localized plasma resonance sensing device according to the present invention; and FIG. 3a is an optical fiber according to the present invention; a schematic diagram of the cladding of a certain region of the optical fiber being completely peeled off; FIG. 3b is a schematic diagram of the optical fiber according to the present invention partially peeled off at a certain region of the optical fiber; 3c and 3d Figure is a schematic cross-sectional view of a fiber according to the present invention partially peeled off in a region of the fiber; Figure 3e is a reference fiber according to the present invention, the surface of the noble metal nanoparticle having a mercaptan head Schematic diagram of modification of a monolayer of a base and a hydroxyl end group; Fig. 3f is a sensing fiber according to the present invention, the surface of the noble metal nanoparticle having a mixed list of a thiol head group and a hydroxyl group or a carboxylic acid end group Schematic diagram of modification of the molecular layer; Fig. 3g is a schematic diagram of the sensing fiber according to the present invention, wherein the surface of the noble metal nanoparticle is modified with a monolayer having a thiol head group and an amino terminal group; Root A schematic diagram of a first embodiment of a self-correcting fiber-type localized plasma resonance sensing device according to the present invention; and a fourth embodiment of a self-correcting fiber-type localized plasma resonance sensing device according to the present invention FIG. 4c is a schematic diagram of a third embodiment of a self-correcting fiber-type localized plasma resonance sensing device according to the present invention; and FIG. 5 is a self-correcting fiber-type localized plasma according to the present invention; A schematic diagram of a second embodiment of a resonance sensing device; FIG. 6a is a schematic diagram of a third embodiment of a self-correcting fiber-type localized plasma resonance sensing device according to the present invention; and FIG. 6b is a reference according to the present invention A schematic diagram of an optical fiber and a sensing fiber whose end faces are respectively modified with a layer of noble metal nanoparticles; FIG. 7 is a schematic diagram of a self-correcting fiber-type localized plasma resonance sensing system according to the present invention; A signal-time relationship diagram obtained by the first embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention; and FIG. 8b is a self-correcting fiber-type localized plasma according to the present invention. a relative signal-time relationship diagram obtained by the first embodiment of the vibration system; FIG. 9a is a signal-time relationship diagram obtained by the second embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention; 9b is a relative signal and log concentration map obtained according to the second embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention; and FIG. 10a is a self-correcting fiber-type localized electric field according to the present invention. a signal-time relationship diagram obtained by the third embodiment of the slurry resonance system; and FIG. 10b is a relative signal-time relationship diagram obtained by the third embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention; Figure 11a is a signal-time relationship diagram obtained in accordance with a fourth embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention; and Figure 11b is a self-correcting fiber-type localization power according to the present invention. A relative signal-time relationship diagram obtained by the fourth embodiment of the slurry resonance system; and Fig. 12a is a signal obtained by the fifth embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention - Figure 12b is a relative signal-time relationship diagram obtained in accordance with the fifth embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention; Figure 12c is a self-correcting type according to the present invention. FIG. 13 is a schematic diagram of a self-correcting type localized plasma resonance sensing device according to a sixth embodiment of the present invention; FIG. 13 is a diagram showing a relationship between a relative signal and a logarithmic concentration obtained by the fifth embodiment of the fiber-optic localized plasma resonance system; Figure 14(A) is a representative sensing diagram showing the normalized response of both the sensing optical waveguide component and the reference optical waveguide component; Figure 14(B) is a diagram showing the compensation response obtained by the sensing system; Figure 15 (A) is a diagram depicting a real-time normalized response of a sensing optical waveguide component and a reference optical waveguide component in response to a pure water sample implanted at about 15 ° C and 45 ° C in a self-correcting localized plasma resonance system; Figure 15 (B) shows a compensation response map obtained by a sensing system in response to a change in temperature; Figure 16 (A) shows both the reference optical waveguide element and the sensing optical waveguide element in order to expose the two waveguide elements sequentially To increase the concentration from 26.2nM to 349nM Data of various samples of biotin antibody dissolved in PBS buffer; FIG. 16(B) shows a compensation response map obtained by the self-correcting type sensing system of the sixth embodiment of the present invention; FIG. 16(C) shows Inject 1X PBS solution into the sensor wafer to establish a flat baseline, then add various red erythrosine (erythrosine, 4×10) dissolved in 10X PBS buffer and spiked with anti-biotin antibody from 26.21 nM to 349 nM. ^-5 g/mL) the solution sample is sequentially injected into the sensor wafer; FIG. 16(D) shows the compensation sensor response diagram of the sixth embodiment of the present invention; and FIG. 17 shows the sixth embodiment of the present invention Calibration curve of avidin antibody obtained by self-corrected sensing system; Figure 18 (A) shows sensing optical waveguide component injected with antibiotic sample of 52.4 nM in 5X PBS (RI=1.33918) And a real-time normalized response map of the reference optical waveguide element; FIG. 18(B) shows a compensation response map obtained by the self-correcting type sensing system of the sixth embodiment of the present invention; and FIG. 19(A) shows a linear transmission (in-line transmission) obtained erythrocyte red solution and reflection table of optical waveguide component The dissipative spectrum of the noble metal nanoparticle layer on the surface; FIG. 19(B) shows the real-time normalized response diagram of the optical waveguide component implanted with the antibiotic sample having a concentration of 52.4 nM in the erythrosine solution; Figure (C) is a diagram showing the compensation response obtained by the self-correcting type sensing system of the sixth embodiment of the present invention.

Please refer to FIG. 2, which depicts a schematic diagram of a self-correcting fiber-type localized plasma resonance sensing device in accordance with the present invention. In the figure, the self-correcting fiber-type localized plasma resonance sensing device 2 includes a reference fiber 21, a sensing fiber 22, and a carrier 23. The reference fiber 21 is modified with a first precious metal nanoparticle layer 211 and receives incident light to produce a first localized plasma resonance sensing signal. The sensing fiber 22 is modified with a second precious metal nanoparticle layer 221. The second precious metal nanoparticle layer 221 is further modified by the identification element 2211 and receives incident light to produce a second localized plasma resonance sensing signal. The carrier 23 is used to place the reference fiber 21 and the sensing fiber 22. The processing unit may refer to the second localized plasma resonance sensing signal according to the first localized plasma resonance sensing signal. The identification element can be a chemical recognition molecule, an antibody, an antigen, a lectin, a hormone receptor, a nucleic acid or a saccharide.

For the reference fiber 21 or the sensing fiber 22, a fiber region whose cladding is completely peeled off can be selected, as shown in Fig. 3a, or a fiber region whose cladding is partially peeled off, as shown in Fig. 3b. A cross-sectional view of the partially peeled fiber of the cladding of the selected region is shown in Figures 3c and 3d. The reference fiber 21 can be modified by the first noble metal nanoparticle layer 211 after stripping the cladding layer, and further modified on the surface of the noble metal nanoparticle with a hydroxyl group-containing (-OH) molecule to provide hydrophilicity. The surface of the first noble metal nanoparticle layer 211 reduces non-specific surface adsorption as shown in Figure 3e. In order to functionalize a molecule having a hydroxyl end group on the surface of the noble metal nanoparticle, a mercaptohexanol (MCH) solution may be prepared, and the reference optical fiber 21 having the first noble metal nanoparticle layer 211 is immersed in the solution. Carry out the reaction.

The sensing fiber 22 is also modified with the second noble metal nanoparticle layer 221 after stripping the cladding layer, and further modifying the specific identification element 2211 on the surface of the noble metal nanoparticle to provide the sensing fiber 22 with a specific detection capability; for example The surface of the noble metal nanoparticles may be functionalized with a long chain thiol molecule containing a carboxylic acid end group (-COOH) or an amine end group (-NH). In order to functionalize the surface of the noble metal nanoparticles with a long-chain thiol molecule containing a carboxylic acid group end group (-COOH) and reduce non-specific surface adsorption, a fluorenyl group having a volume ratio of 1:4 can be used in the self-assembly reaction. A solution of undecic acid (MUA) and mercaptohexanol (MCH) as shown in Figure 3f. By adding a short carbon chain MCH molecule in a specific ratio, the distance between individual molecular probes can be spatially dispersed, and the steric hindrance in antibody-antigen identification can be eliminated to increase the recognition efficiency. Further, in order to functionalize the surface of the noble metal nanoparticles with a thiol molecule having an amino group-containing end group, as shown in Fig. 3g, it may be configured with a cystamine solution, and the second noble metal nanoparticles may be modified. The method of immersing the sensing fiber 22 of layer 221 in this solution for reaction is performed.

Please refer to FIG. 4, which shows a first embodiment of a self-correcting fiber-type localized plasma resonance device according to the present invention. When the fiber-type localized plasma resonance device 2 is a microfluidic wafer, The channel portion can be designed according to the requirements of the sample to be tested, and the factors to be considered include the fluid mechanics, surface tension, fluid volume, internal pressure and sample residue after the sample entering. In addition, internal and external factors caused during chip packaging can be one of the requirements. Please refer to Figure 4a, which shows a schematic of a basic microfluidic wafer. In the microfluidic wafer, a single sample cell 41 for placing the reference fiber 21, the sensing fiber 22, and a sample having a volume substantially less than or equal to 50 microliters is suitably designed. Referring to Figure 4b, there is shown a schematic diagram of a split flow microfluidic wafer in accordance with the present invention. In the figure, the sample flows to two microfluidic channels, and the volume of the sample is substantially less than or equal to 40 microliters. Referring to FIG. 4c, a schematic diagram of a multiple detection type microfluidic wafer is shown in which the reference fiber 21 can be placed in synchronization with a plurality of sensing fibers 22 having different identification elements for multiple sensing. The sample volume required for the test is approximately 20-80 μl, which reduces sample consumption to meet the needs of microanalytical detection and further provides the ability to simultaneously test multiple analytes to save time.

Referring next to Fig. 5, there is shown a schematic view of a second embodiment of a self-correcting fiber-type localized plasma resonance device in accordance with the present invention. When the fiber-type localized plasma resonance device 2 is used in the micro-sample tray, the reference fiber and the sensing fiber may be disposed on the distal end surface of the fiber with the mirror 51 to reflect the first localized plasma resonance sensing signal and the first Two localized plasma resonance sensing signals. The filter membrane 52 may be further provided to block interference substances having a size larger than the average pore diameter of the membrane outside the filter membrane 52.

Next, please refer to Fig. 6a, which shows a schematic view of a third embodiment of a self-correcting fiber-type localized plasma resonance device according to the present invention. In the detection of the actual environment or biological sample or specimen, the sampling device can be immersed or penetrated into a specific sample or target to complete the in-situ detection. Therefore, the sensing device is suitable for medical treatment. Equipment for in vivo or current sampling and analysis. When detecting a real sample, various interfering substances may be present in the sample, so a filter 62 may be added to the outer layers of the reference fiber 21 and the sensing fiber 22 to separate interfering substances having a size larger than the average pore diameter of the film. It is outside the filter membrane 62; in addition, a rigid bracket 61 provided with a hole in the bracket can be placed to enhance the physical strength of the entire sensor. In addition, the noble metal nanoparticle layers 211, 221 may be modified on the end faces of the reference fiber 21 and the sensing fiber 22, respectively, to facilitate in situ sampling and analysis, as shown in FIG. 6b.

Referring now to Figure 7, there is shown a schematic diagram of a self-correcting fiber-type localized plasma resonance sensing system in accordance with the present invention. The self-correcting fiber-type localized plasma resonance sensing system is characterized by comprising: a light source 71, a fiber-type localized plasma resonance sensing device 72, and a light detecting unit 73. Light source 71 can be a light emitting diode (LED) for generating incident light, wherein the incident light is coupled to fiber-optic localized plasma resonance sensing device 72 through a fiber coupler. The fiber-optic localized plasma resonance sensing device 72 includes a reference fiber 721, a sensing fiber 722, and a carrier 723. The reference fiber 721 is modified with a first layer of noble metal nanoparticles and receives incident light to produce a first localized plasma resonance sensing signal. The sensing fiber 722 is modified with a second layer of noble metal nanoparticles, wherein the second layer of noble metal nanoparticles is further modified with an identification element and receives incident light to produce a second localized plasma resonance sensing signal. The carrier 723 is used to place the reference fiber 721 and the sensing fiber 722. The light detecting unit 73 can be a photodiode for receiving the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal. The processing unit 74 can refer to the second localized plasma resonance sensing signal according to the first localized plasma resonance sensing signal. The fiber-optic localized plasma resonance sensing system further includes a lock-in amplifier 75 and a signal generator 76, wherein the lock-in amplifier 75 can amplify the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal. The system noise is suppressed, and the signal generator 76 drives the light source to generate and adjust the incident light and provide a reference signal to the lock-in amplifier.

When performing a biological or chemical sample detection, it is possible to perform different concentrations of sensing operations using the selectivity of the identification elements, wherein the sensing fiber 722 is modified with an identification element, while the reference fiber 721 is absent. The dielectric environment in the vicinity of the sensing fiber 722 changes according to the interaction between the identification element on the surface of the noble metal nanoparticle and the object to be tested, thereby reducing the second localized plasma resonance sensing signal, and the time signal generated is generated. Molecular binding kinetics curve. Since the surface of the reference fiber 721 is not modified by the identification element, the first localized plasma resonance sensing signal change is only due to a change in the refractive index of the sample, non-specific adsorption, or environmental factors. The first-domain plasma resonance sensing signal may be a signal I _R0 obtained by detecting a blank solution and the surface of the reference nano-particle of the optical fiber 721 is not modified by the identification element, and detecting different concentrations of the sample to be tested by referring to the optical fiber 721 The obtained signal I _R ; the second localized plasma resonance sensing signal can be divided into a signal I _{S0 for} detecting the blank solution and sensing the surface of the nanoparticle of the optical fiber 722 to be modified by the identification element, and by sensing the optical fiber 722 The signal I _{S from the} sample is detected. Please refer to the following formula: I' ₀ = I _S0 /I _R0

I'=I _S /I _R

T'=I'/I' ₀ =(I _S /I _R )/(I _S0 /I _R0 )=(I _S /I _S0 )/(I _R /I _R0 )=T _S /T _R

The parameters used in the above formula are described in the following: I _'0 is detected when the same blank solution, the above _S0 I I obtained by dividing the correction signal _R0; I' is the same sample to detect the above-described divided by I _R I _S the resulting corrected signal; and T '= I' / I ' 0 represents the relative signal obtained after the self-calibration. The analyte concentration value is taken as the -log x-axis, and with respect to the axis a T y '= I' / I _'0 graphically on FIG linear portion between the opposite signal as the -log concentration correction chart.

Please refer to Fig. 8a, which shows a signal-time relationship diagram obtained in the first embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention. Referring also to Fig. 8b, there is shown a relative signal-time relationship of the first embodiment of the self-correcting fiber-type localized plasma resonance system in accordance with the present invention. Due to the sensitivity of the localized plasma resonance to ambient temperature, the self-correcting fiber-type localized plasma resonance sensing system according to the present invention performs individual tests on the reference fiber 81 and the sensing fiber 82 at different temperatures simultaneously. Self-correcting effects of temperature changes are verified by self-correcting operations. In general, the refractive index of a solution is temperature dependent; therefore, the signal drops as the temperature rises and vice versa. In a self-correcting fiber-optic localized plasma sensing system, when the temperature rises, the signals in both the reference fiber and the sensing fiber signal decrease simultaneously; and when the temperature drops, the signals rise together; thus, the relative signal is utilized ( I'/I' ₀ ) A signal obtained by self-correction can observe a relatively flat signal on the time map.

Referring to Figure 9a, there is shown a signal-time relationship diagram obtained in accordance with a second embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention. See also Figure 9b, which shows a relative signal-log concentration plot obtained in accordance with a second embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention. In order to confirm the feasibility of the system for qualitative and quantitative, the experiment of using vitamin H as an identification element to detect different concentrations of streptavidin was designed. It can be seen from the experimental results that when a high concentration of avidin is driven, it can be seen that there is no change in the signal from the reference fiber 91, mainly because of the hydrophilicity inhibition of the hydroxyl end group (-OH) on the surface of the noble metal particle. Non-specific surface adsorption; at the same time, when the sensing fiber 92 is functionalized with biotin and combined with biotin and avidin, the signal degradation is evident (as shown in Figure 9a). The resulting time signal exhibits a molecular binding kinetic curve. Using the above results, the signal-log concentration map has a correction coefficient close to 0.996 of 0.990 obtained from a non-self-correcting single fiber sensing system by sequentially injecting different concentrations of avidin for detection (eg, 9b). The figure also shows a detection limit of 3.8 × 10 ^-11 M, which is also similar to 4.1 × 10 ^-11 M derived from a non-self-correcting single fiber sensing system.

Referring to Fig. 10a, there is shown a signal-time relationship diagram obtained in accordance with a third embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention. Please also refer to FIG. 10b, which shows a relative signal-time relationship obtained in accordance with a third embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention. During the test, a highly viscous sample may cause a change in the refractive index in the solution to cause an error during the test. For example, the detection of IL-1β content in the joint fluid of patients with osteoarthritis (OA) is provided, wherein the joint fluid sample still exhibits a high viscosity after the sample preparation treatment. However by according to the invention The self-correcting sensing system allows for detection without the need for excessive dilution steps, thus helping to increase detection limits.

A solution of the MUA/MCH mixture is used which is capable of self-assembling a mixed monolayer molecular film on the surface of the gold nanoparticles. The method of forming a probe comprises the steps of first activating the carboxyl end group of MUA and then conjugated it to an antibody against anti-human IL-1β via a chemical reaction. In a traditional single-fiber sensing system, when testing a real sample of joint fluid, it is necessary to first dilute the sample with high viscosity, which may introduce errors during dilution, resulting in inaccuracy and time consuming, and also reduce the method. Detection limit.

When the real sample of the joint fluid is detected by the self-correcting type fiber-optic localized plasma sensing system according to the present invention, the detection can be started only after slightly diluting the viscous joint fluid. Since the ultimate goal of this system is to determine the amount of IL-1β in the knee joint fluid of patients with osteoarthritis, it can be seen that the injection of a viscous sample causes an initial sharp drop in the signal from the reference fiber and the sensing fiber (eg, As shown in Fig. 10a, the signal that drops so sharply is an error, resulting in an error. Self-correcting by the self-correcting fiber-type localized plasma sensing system, it can be seen that there is no characteristic molecular binding kinetic curve of the initial sharply falling signal (as shown in Figure 10b), and the measured IL in the sample. The -1 beta concentration is 1.72 x 10 ^-10 M, which is similar to the results obtained with a single fiber sensing system, but does not require excessive dilution.

In the following, reference is made to Fig. 11a, which shows a signal-time relationship diagram obtained in accordance with a fourth embodiment of the self-correcting fiber-type localized plasma resonance system of the present invention. Please also refer to FIG. 11b, which shows a relative signal-time relationship diagram in the fourth embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention. When a real sample exhibits certain colors, spectral interference may occur, causing errors in the analysis. In an experiment using a juice containing CymMV orchid virus, the orchid juice was still dark blue after dilution (the absorption band was at about 600 nm), which may cause spectral interference and a difference in refractive index of the solution compared to the blank solution. When the juice with the CymMV virus is injected, since the reference fiber is only modified with MCH and there is no antibody specific for the virus, it can be seen from Fig. 11a that the initial sharp drop from the reference fiber 111 signal, mainly It is caused by spectral interference and refractive index change in the solution; and the sensing fiber 112 is modified to have specificity for the virus, so after the initial sharp drop of the signal, the molecular binding occurs due to the interaction between the antibody and the antigen. Kinetic curve. Using two signals from the reference optical fiber 111 and the sensing fiber _{112 (I '/ I' 0} ) of the self-calibration signal, this is evident, as shown in FIG. 11b of the acquired through self-calibration of the sensing system The data provides correction characteristics for only the molecular binding kinetics curve, thereby reducing the spectral interference that occurs when the sample is rendered colored.

Next, please refer to Fig. 12a, which shows a signal-time relationship diagram obtained in the fifth embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention. Referring then to Figure 12b, there is shown a relative signal-time relationship diagram for a fifth embodiment of a self-correcting fiber-type localized plasma resonance system in accordance with the present invention. In addition, please refer to Fig. 12c, which shows a relative signal-log concentration relationship diagram in the fifth embodiment of the self-correcting fiber-type localized plasma resonance system according to the present invention. Usually, several biochemical substances of interest can coexist in a real sample. In order to be able to detect such biochemical substances at the same time, an element for multiplex detection is designed, which is provided with a reference fiber without an identification element and a plurality of sensing fibers, wherein each of the plurality of sensing fibers is sensed Each of the optical fibers is modified by one of the identification elements of the biochemical substances, so that self-correction can be used for simultaneous multi-detection.

An experiment for multiplex detection by means of a self-correcting fiber-type localized plasma sensing system is to detect a solution consisting of avidin and anti-DNP antibody (anti-DNP) with different concentrations of avidin and anti-DNP and self-calibration based on the reference fiber to achieve multiple detections. Figure 12a shows a signal-time diagram from the reference fiber and the sensing fiber. It can be seen that the avidin detected by the biotin functionalized fiber at different concentrations and the fiber detected by the DNP functionalized fiber Molecular binding dynamics map of anti-DNP. Using the self-correction of the signal using the signal from the reference fiber 111 and the sensing fiber, it can be seen that as the concentration of the analyte increases, the characteristic molecular binding kinetic curve and the linear calibration curve are still observed, confirming the self-correcting simultaneous multiplicity The feasibility of the test is shown in Figures 12b and 12c.

However, the reference fiber and the sensing fiber are taken as examples, and the present disclosure is not limited thereto. In fact, the self-correcting fiber-type localized plasma sensing device can be an example of a self-correcting type localized plasma resonance sensing device. The reference fiber can be an example of a reference optical waveguide component, and the sensing fiber can be An example of sensing an optical waveguide component. The optical waveguide component can be selected from one of the following: an optical fiber, a channel waveguide, a planar waveguide, or a tubular waveguide.

Referring now to Figure 13, there is shown a schematic diagram of a self-correcting type localized plasma resonance sensing device in accordance with a sixth embodiment of the present invention. The correction slope of the reference optical waveguide element 31 and the sensing optical waveguide element 32 of the sixth embodiment may be different. Processing unit 74, which may be a processor, utilizes the difference between the signal generated by sensing optical waveguide component 32 and the signal generated by reference optical waveguide component 31 to obtain a sensor response. Here, the self-correcting type localized plasma resonance sensing device and other elements of the system in other embodiments may exist in the sixth embodiment. Further, materials of the elements of the other embodiments can be used in the elements of the sixth embodiment.

In more detail, the self-correcting type localized plasma resonance sensing device of the sixth embodiment includes the reference optical waveguide element 31, the sensing optical waveguide element 32, and the carrier 33. The carrier 33 places the reference optical waveguide element 31 and the sensing optical waveguide element 32. Here, the carrier 33 has a channel 34 for placing the reference optical waveguide element 31 and the sensing optical waveguide element 32. In addition, the carrier has a V-shaped channel 35 connected to the channel 34 for injecting the sample into the channel 34. Here, an opening 37 is formed at the top of the V-shaped channel 35 for injecting a sample into the V-shaped channel 35, and both ends of the V-shaped channel 35 are respectively connected to the channel 34 to take the sample from the V shape Channel 35 guide It enters the channel 34. Here, the channel 34 has openings 36 corresponding to the reference optical waveguide element 31 and the sensing optical waveguide element 32, respectively, for flowing out of the sample. Further, the reference optical waveguide element 31 is modified with a first noble metal nanoparticle layer. Part of the incident light emitted from the light source 71 is introduced into the reference optical waveguide element 31 to generate a first localized plasma resonance sensing signal. Here, the reference optical waveguide element 31 has a first correction slope. The first-domain plasma resonance sensing signal includes a first signal generated by detecting the blank solution by the reference optical waveguide element 31 and a second signal generated by detecting the sample with the reference optical waveguide element 31.

Further, the sensing optical waveguide element 32 is modified with a second noble metal nanoparticle layer. The second precious metal nanoparticle layer is further modified with an identification element. Here, the identifier includes a chemical recognition molecule, an antibody, an antigen, a lectin, a hormone receptor, a nucleic acid, or a saccharide. Another portion of the incident light emitted from the source 71 is directed into the sensing optical waveguide component 32 to produce a second localized plasma resonant sensing signal. Here, the sensing optical waveguide element 32 has a second correction slope that is different from the first correction slope of the reference optical waveguide element 31. The second localized plasma resonance sensing signal includes a third signal generated by sensing the optical waveguide component 32 to detect the blank solution and a fourth signal generated by sensing the optical waveguide component 32 to detect the sample. Here, the number of each of the reference optical waveguide element 31 and the sensing optical waveguide element 32 is one or more.

The light detecting unit 73, for example, a composition of at least one photodiode, receives the first localized plasma resonance sensing signal and the second localized plasma resonance sensing generated by the reference optical waveguide component 31 and the sensing optical waveguide component 32, respectively. Signal. The light detecting unit 73 then transmits the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal to the processing unit 74, which can be a processor. The processing unit 74 normalizes the first difference between the second signal and the first signal and normalizes the second difference between the fourth signal and the third signal. Processing unit 74 then adjusts the normalized second difference using the first corrected slope of reference optical waveguide element 31 and the second corrected slope of sensed optical waveguide element 32. Therefore, the adjusted second difference is obtained. Then, the processing unit 74 uses the difference between the normalized first difference and the adjusted second difference to obtain The sensor responds. Therefore, it is possible to easily and more accurately compensate for temperature effects and bulk-composition effects as well as non-specific adsorption. Further, the characteristics of the reference optical waveguide element 31 and the sensing optical waveguide element 32 may be different.

For example, the localized plasma resonance sensing technique used in the present invention is based on the absorption of an evanescent wave by a layer of noble metal nanoparticles on an optical waveguide element. When light is transmitted along the optical waveguide element through continuous total internal reflection (TIR), the noble metal nanoparticles are excited by the evanescent field, and thereby the light passing through the optical waveguide element is interacted with the noble metal nanoparticle. Attenuate by action. When this attenuation is increased by multiple total internal reflections, the low absorption amount of the noble metal nanoparticles can be greatly increased. Because the localized plasma resonance sensing technology is based on the change in the absorption of the gold nanoparticle layer in different refractive index (RI) environments, it can be assumed that the refractive index of the medium surrounding the noble metal nanoparticle layer increases by Δn. At the time, the bulk absorption coefficient of the noble metal nanoparticle layer can be increased from α _{0 of the} blank solution to α ₀ +Δα of the sample. Therefore, if a plot of Δα/α ₀ versus Δn is plotted, a linear regression line with a slope of m will be obtained. Therefore, the sensor response of localized plasma resonance can be approximated by the following relationship: ΔI/I ₀ = (I ₀ -I) / I ₀ =1 - I / I ₀ Δα/α ₀ , wherein the normalized response ΔI/I ₀ is defined as the intensity of the signal collected from the optical waveguide component of the modified noble metal nanoparticle layer immersed in the sample (I) corresponding to the immersion in the blank solution The signal intensity collected by the same optical waveguide component of (I ₀ ), and the plot of ΔI/I ₀ relative to Δn will also produce a linear regression line with a slope of m. Since the molecules bound to the surface of the noble metal nanoparticles will promote an increase in the regional refractive index in the sensing depth of the noble metal nanoparticles, this combination will cause a decrease in the transmitted light intensity through the optical waveguide elements. Therefore, by using the recognition molecules conjugated to the surface of the noble metal nanoparticles, the corresponding analyte can be sensed without using the label.

Temperature effects and batch composition effects can cause significant changes in the response of the localized plasma resonance sensor, which can conceal the response changes produced by a particular combination of analytes. Here, a new self-correcting mechanism in the dual channel localized plasma resonance sensing system is used to compensate for such effects in the composite sample (eg, thermal, discrete refractive index, color interference, etc.). The sensory optical waveguide component 32, which has a specific identification molecule immobilization, will measure the specific combination of analytes and unwanted temperature effects and batch composition effects. Ideally, there should be no non-specific adsorption of the sensing optical waveguide component 32. However, in essence, it is difficult to completely avoid non-specific adsorption. Therefore, the compensation response can be described as the following relationship: Where ΔI _S /I _S0 is the normalized response of the sensing optical waveguide component 32, m _S is the corrected slope of the sensing optical waveguide component 32, Δn _M is the different refractive index change of the medium, and Δn _{NA is the} non-specific adsorption site The resulting regional refractive index change, if any, Δn _SA is the effective regional refractive index change due to specific adsorption.

In addition, an immobilized reference optical waveguide element 31 without a specific identification molecule will only measure temperature effects and batch composition effects as well as non-specific adsorption, if any, and sensor response, which can be described as Relationship: Where ΔI _R /I _R0 is the normalized response of the reference optical waveguide element 31 and m _R is the corrected slope of the reference optical waveguide element 31.

After rewriting formula (1), only the sensor response caused by specific adsorption can be expressed as: Ideally, if the corrected slope of the sensed optical waveguide component 32 is equal to the corrected slope of the reference optical waveguide component 31 (i.e., m _S = m _R ), then from equations (2) and (3),

However, strict quality control is followed to control the surface coverage of the noble metal nanoparticles on the waveguide surface and α ₀ , and thus the correction slope between the sensing optical waveguide component 32 and the reference optical waveguide component 31 is nearly identical It is very costly and cumbersome.

And if the correction slope of the sensing optical waveguide element 32 is different from the correction slope of the reference optical waveguide element 31, that is, m _S ≠m _R , then from equations (2) and (3),

When the injection of the sample causes a change in the refractive index of the medium and Δn _M , the normalized responses of the sensing optical waveguide component 32 and the reference optical waveguide component 31 will be ΔI _{S, M} /I _S0 and ΔI _{R, M} /I, respectively. _R0 . These changes can be easily learned from the sensing map. ΔI _{S, M} /I _S0 and ΔI _{R, M} /I _R0 versus Δn _M can respectively indicate the correction slope of the sensing optical waveguide element 32 and the reference optical waveguide element 31, and the equation (5) can be rewritten as

This result means that deliberate use of blank solutions and samples with different refractive indices will more easily and accurately compensate for thermal effects, batch composition effects, and non-specific adsorption.

In order to quantify the sensor response of the self-correcting localized plasma resonance sensing system corresponding to different refractive index changes, pure water and phosphate buffered saline (PBS) solution with different refractive indices (1.33250-1.35726) will depend on The order is injected into the sensor wafer. Referring to Fig. 14(A), it is shown that the normalized response representative sensing map of the sensing optical waveguide component and the reference optical waveguide component. The baseline is established when the two optical waveguide elements are in contact with pure water. From the baseline of the self-sensing optical waveguide component, the power stability or relative standard deviation of the noise (σ) is estimated to be 0.0073% per 120 seconds. Containing noise caused by five injections of pure water samples, the sum of the residual squared sums (SSE) of the normalized response regression of the sensed optical waveguide elements is estimated to be 0.0319%. As the refractive index of the injected sample is gradually increased, the normalized response of the two optical waveguide elements shows a tendency to step. Through the correction of equation (5), the compensated response should ideally be a straight line with a mean value of zero. Referring to Figure 14 (B), The compensation response obtained by the sensing system is displayed. As shown in Figure 14 (B), the SSE estimate for the regression of the post-compensation response is 0.0151%. This comparable and even better SSE value indicates that the self-correcting localized plasma resonance sensing system effectively compensates for the change in refractive index of the underlying background.

Further, the temperature effect is compensated by using the self-correcting type localized plasma resonance sensing device of the present invention and its system. In general, the refractive index of a solution is related to temperature. It can be found that at a wavelength of 632.8 nm, the change in temperature causes a change in the refractive index of water of 9 × 10 ^-5 RIU. °C ^-1 . Referring to Fig. 15(A), a real-time normalized response map of the sensed optical waveguide component and the reference optical waveguide component in water in response to injection of a pure water sample at about 15 ° C and 45 ° C is depicted. In the self-correcting localized plasma resonance sensing system, when the temperature is lowered, the normalized response of the two optical waveguide components will rise simultaneously; and when the temperature rises, the normalized response of the two optical waveguide components will decrease simultaneously. . Based on the data shown in Fig. 15(A), the SSE estimate for the regression of the normalized response of the sensing optical waveguide element including the temperature effect is 0.2316%. Further, referring to Fig. 15(B), it shows the compensation response obtained by the sensing system in response to the temperature change. As shown in Figure 15 (B), the SSE estimate for the regression of the post-compensation response is 0.0284%. This result shows that the self-correcting localized plasma resonance sensing system provides superior compensation for temperature changes. On the other hand, temperature variation compensation of surface plasmon resonance (SPR) sensors is a challenge.

Furthermore, the non-specific adsorption effect is compensated by using the self-correcting type localized plasma resonance sensing device of the present invention and its system. For biosensor applications of real samples, non-specific adsorption will result in errors in the biosensor based on detection of changes in refractive index adjacent to the surface of the biosensor. Although various methods have been employed to reduce non-specific adsorption of the sensor surface, completely eliminating this effect remains a technical challenge. Instead, the non-specific adsorption effect is compensated by the mechanism of the reference sensor. If the reference sensor is identical except for the non-functionalized recognition molecule, the reference sensor can be used to compensate for non-specific adsorption. Referring to Fig. 16(A), it is shown that the reference optical waveguide element and the sensing optical waveguide element are sequentially exposed to the two waveguide elements to various samples in which the avidin is dissolved in the PBS buffer and has a concentration increased from 26.2 nM to 349 nM. data. As shown in Fig. 16(A), when the sensing optical waveguide element 32 generates a large signal, the reference optical waveguide element 31 will have a small amount of non-specific adsorption. Referring to Fig. 16(B), the compensation response obtained by the self-correcting type sensing system of the sixth embodiment is shown. By the formula (4), the post-compensation response can be calculated by the pairwise difference of the normalized responses between the optical waveguide elements. Referring to Fig. 17, there is shown a calibration chart of the anti-biotin antibody obtained by the self-correcting type sensing system of the sixth embodiment. As shown in Figure 17, the calibration curve for ΔI _S,SA /I _S0 versus logarithmic anti-biotin antibody concentration is linear in the concentration range of 52.4 nM to 349 nM (correlation coefficient r =0.9989, n =3) ). The self-corrected localized plasma resonance sensing system has a detection limit (LOD) of 29.5 nM for avidin antibodies. On the other hand, if the calibration curve is established using only the normalized response of the sensing optical waveguide component 32, the error will be as large as 46%.

Furthermore, the discrete refractive index effect and the non-specific adsorption effect are compensated by using the self-correcting type localized plasma resonance sensing device of the present invention and its system. For example, the refractive index of the buffer used to establish the baseline and dissolve the sample is intentionally chosen to be different to demonstrate the feasibility of compensating for the discrete refractive index effect and the non-specific adsorption effect, and also to obtain the refraction through two optical waveguide sensors. The advantage of the sharp response of the rate change is corrected for the difference in the correction slope between the sensing optical waveguide element 32 and the reference optical waveguide element 31. Referring to Fig. 18(A), the real-time normalized response of the sensed optical waveguide element 32 and the reference optical waveguide element 31 injected into the 5X PBS (RI = 1.33918) containing the avidin sample at a concentration of 52.4 nM is shown. As shown in Fig. 18(A), the normalized response of both the sensing optical waveguide element 32 and the reference optical waveguide element 31 increases immediately and synchronously due to an increase in the refractive index. Thereafter, the normalized response of the sensing optical waveguide component 32 is continuously increased and follows the molecular binding dynamic curve, while the normalized response of the reference optical waveguide component 31 is very slowly increased due to non-specific adsorption. The normalized response due to matrix changes, ΔI _{S, M} /I _S0 and ΔI _{R, M} /I _R0 can be simply achieved by using the molecular binding dynamics curve and the intersection of the response curves due to discrete refractive index changes, It is determined by estimating ΔI _{S, M} and ΔI _{R, M.}

Continuing to the next, referring to Fig. 18(B), the compensation response obtained by the self-correcting type sensing system of the sixth embodiment of the present invention is shown. As shown in Figs. 18(A) and 18(B), the region of the sharp rise in the normalized response provides a mechanism to correct the difference in the correction slope between the sensing optical waveguide element 32 and the reference optical waveguide element 31. The compensated response can be calculated by equation (6). From the calibration curve as shown in Fig. 18(B), the avidin antibody concentration in the doped sample was estimated to be 54.1 ± 4.0 nM ( n = 3), which resulted in a recovery of 103.2%. This result indicates that the self-correcting localized plasma resonance sensing system provides excellent compensation for both the discrete refractive index effect and the non-specific adsorption effect in the sample.

Furthermore, color interference is compensated by using the self-correcting type localized plasma resonance sensing device of the present invention and its system. Referring to Fig. 19(A), there is shown a dissipative spectrum of a ruthenium red solution obtained by linear transmission and a noble metal nanoparticle layer on a reflecting surface of an optical waveguide element. As shown in Fig. 19(A), when the real sample has a color overlapping the excitation spectrum of the incident light, the evanescent wave at the reflective surface of the waveguide element can be absorbed by the matrix of the real sample and the noble metal nanoparticles. Therefore, spectral interference will occur and result in analysis errors. In the example of the present invention, a colored substrate is produced by adding a dye (erythrosine) to the sample. Obviously, the peak wavelength of the erythrocyte solution (2×10 ^-5 g/mL) at about 527 nm overlaps with the peak wavelength emitted by the LED used in this example, and also with the noble metal nanoparticle layer. The plasma absorption bands overlap.

Referring to FIGS. 19(B) and 19(C), respectively, showing a real-time normalized response of an anti-biotin antibody sample containing a concentration of 52.4 nM in an infusion solution of an optical waveguide element, and a sixth embodiment of the present invention The compensation response obtained by the self-correcting sensing system of the example; as shown in Fig. 19 (B) and Fig. 19 (C), when the anti-biotin antibody (52.4 nM) is injected into the erythrosine solution In the case of the wafer, the normalized response of the sensing optical waveguide component 32 and the reference optical waveguide component 31 is increased immediately and simultaneously due to color interference. On the other hand, after the signal is rapidly increased, the normalized response of the sensing optical waveguide component 32 is continuously increased and follows the molecular bonding dynamic curve, and the normalized response of the reference optical waveguide component 31 is very slow due to non-specific adsorption. Increase in land. For the same reason, the compensated sensor response can be calculated by equation (6). According to the calibration curve shown in Figure 17, the concentration of the avidin antibody in the doped sample is estimated as 49.2 ± 5.2 nM (n = 3), which yielded a recovery of 93.9%. This result indicates that the self-correcting localized plasma resonance sensing system provides excellent compensation for color interference in the sample matrix.

In addition, the anti-biotin antibody in the composite medium can be directly detected. In more detail, many real samples (such as whole blood, plant sap) have very complex matrices that lead to direct biosensing in terms of sample blank solution, color interference, and refractive index differences between non-specific adsorptions. The result is an error. However, in the present invention, a simulated composite sample composed of a high refractive index buffer and erythrosin as a color interference substance is used herein to confirm that the self-correcting type localized plasma resonance sensing system is used for direct detection. The feasibility of anti-biotin antibodies in such composite samples. As shown in Fig. 16(C) and Fig. 16(D), respectively, it was shown that a 1X PBS solution was implanted in the sensor wafer to establish a flat baseline, and then dissolved in 10X PBS buffer and mixed with concentration from 26.21 nM of various erythrosine (4 x 10 ^-5 g / mL) solution samples of 349 nM anti-biotin antibody were sequentially injected into the sensor wafer, and the compensation sensor response of the sixth embodiment of the present invention . The calibration curve of ΔI _S,SA /I _S0 versus logarithmic antibiotic antibody concentration is shown in Fig. 17, which has a linear relationship in the range of 52.4 nM-349 nM (correlation coefficient r = 0.9993, n = 3). The self-corrected localized plasma resonance sensing system has an LOD line of 31.8 nM for the avidin antibody in this composite medium, which approximates the LOD of the avidin antibody in 1X PBS (LOD = 29.5 nM). To compare the results from samples in simple buffers and in composite solutions, statistical analysis of these two sets of results was performed by a pair-t test. The results show that the average of the two groups meets the 95% mutual confidence interval, which means that the self-correcting localized plasma resonance sensing system provides a simple, reproducible and high sensitivity method to sense the composite. The analyte in the sample.

Therefore, in the present invention, a new method is demonstrated which performs plasma resonance biosensing based on a self-correcting double channel localized plasma resonance sensing system. This method uses the reference optical waveguide component 31 to accurately compensate for systematic errors (e.g., temperature variations, discrete refractive index changes, color interference, and/or non-specific adsorption) of the sensed optical waveguide components in a single microfluidic wafer. The self-correcting mechanism may allow for immediate localized plasma resonance biosensing to be applied outside of the laboratory where room temperature conditions may not be maintained. And it is also special Suitable for biosensing in composite real samples, the interference effects in composite real samples pose a significant challenge to many unlabeled refractive index biosensors. Since localized plasma resonance sensors are based on normalized responses for data analysis, the need for precise optical alignment can be mitigated. Therefore, with the self-correction mechanism to compensate for thermal effects, batch composition effects, and non-specific adsorption, the dual-channel localized plasma resonance sensing system has the potential to be developed for environmental, agricultural, food, and health care. The field of biochemical sensors for chemical and biochemical analysis of the field.

The above description is merely illustrative and not restrictive. All of the valid equivalent modifications, changes, or conversions made without departing from the spirit and scope of the invention are considered to be covered by the scope of the invention.

2‧‧‧Self-corrected fiber-optic localized plasma resonance sensing device

21‧‧‧Reference fiber

211, 221‧‧‧ precious metal nanoparticle layer

22‧‧‧Sensing fiber

2211‧‧‧ Identification element

23‧‧‧ Carrier

Claims (23)

A self-correcting type localized plasma resonance sensing device, comprising: a reference optical waveguide component modified with a first noble metal nanoparticle layer, and a portion of incident light is introduced into the reference optical waveguide component to generate a a first-domain plasma resonance sensing signal, wherein the first localized plasma resonance sensing signal comprises a first signal generated by detecting a blank solution by the reference optical waveguide component and detected by the reference optical waveguide component The product generates a second signal, wherein the reference optical waveguide component has a first correction slope; a sensing optical waveguide component modified with a second noble metal nanoparticle layer, the second noble metal nanoparticle layer An identification element is further modified, and another portion of the incident light is introduced into the sensing optical waveguide component to generate a second localized plasma resonance sensing signal, wherein the second localized plasma resonant sensing signal comprises the sensing light The waveguide component detects a third signal generated by the blank solution and a fourth signal generated by the sensing optical waveguide component detecting the sample, wherein the sensing light wave The component has a second correction slope, wherein a processor normalizes a first difference between the second signal and the first signal and normalizes a second difference between the fourth signal and the third signal a value, wherein the processor adjusts the normalized second difference using the first corrected slope and the second corrected slope to obtain an adjusted second difference, and then the processor utilizes normalized a difference between the first difference and the adjusted second difference to obtain a sensor response; and a carrier that places the reference optical waveguide component and the sensing optical waveguide component. The self-correcting localized plasma resonance sensing device of claim 1, wherein the first noble metal nanoparticle layer is modified on a reflective surface of the reference optical waveguide component. The self-correcting localized plasma resonance sensing device of claim 1, wherein the second noble metal nanoparticle layer is modified on a reflective surface of the sensing optical waveguide component. The self-correcting localized plasma resonance sensing device of claim 1, wherein the reference optical waveguide component and the sensing optical waveguide component are an optical fiber, a channel waveguide, a planar waveguide, or a tubular waveguide. The self-correcting localized plasma resonance sensing device of claim 4, wherein the first noble metal nanoparticle layer is stripped to one of the optical fibers when the reference optical waveguide component is an optical fiber Modifications are made at the area or at one end. The self-correcting type localized plasma resonance sensing device according to claim 4, wherein when the sensing optical waveguide component is an optical fiber, the second noble metal nanoparticle layer is in a cladding stripping region of the optical fiber. Or modify at one end. The self-correcting localized plasma resonance sensing device according to claim 1, wherein the self-correcting type localized plasma resonance sensing device is a microfluidic wafer or a current sampling and analyzing device. The self-correcting type localized plasma resonance sensing device according to claim 7, wherein the reference optical waveguide component and the sensing optical waveguide component are respectively at an end surface of the reference optical waveguide component and the sensing optical waveguide component One of the end faces constructs a mirror surface, wherein the reference optical waveguide component and the sense optical waveguide component are optical fibers or tubular waveguides. The self-correcting type localized plasma resonance sensing device of claim 8, wherein the reference optical waveguide component and the sensing optical waveguide component are further provided with a filter film and a rigid bracket having at least one opening, The mirror surface is configured to reflect the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal, and the filter filters an interference substance having a size larger than an average aperture of the filter, and the Rigid bracket wraps the ginseng The optical waveguide component and the sensing optical waveguide component are used to increase the mechanical strength of the self-correcting localized plasma resonance sensing device during the sampling operation. The self-correcting localized plasma resonance sensing device according to claim 1, wherein the identification element comprises a chemical recognition molecule, an antibody, an antigen, a lectin, a hormone receptor, a nucleic acid or a saccharide. The self-correcting localized plasma resonance sensing device of claim 1, wherein the sensor response is represented by the following formula: Wherein ΔI _{S, SA} represents the sensor response caused by only specific adsorption, and I _S0 represents the third signal generated by the sensing optical waveguide component detecting the blank solution, and ΔI _S represents the fourth signal and the The second difference between the third signals, ΔI _{S, M} /I _S0 and ΔI _{R, M} /I _R0 respectively represent the normalized response of the sensing optical waveguide component and the reference optical waveguide component, which respectively represent The second correction slope and the first correction slope, ΔI _R represents the first difference between the second signal and the first signal, and I _R0 represents the detection of the blank solution by the reference optical waveguide component The first signal. A self-correcting type localized plasma resonance sensing system, comprising: a light source that generates an incident light; and a certain domain plasma resonance sensing device, comprising: a reference optical waveguide component, which is a first noble metal The rice particle layer is modified, and a portion of the incident light is introduced into the reference optical waveguide component to generate a first localized plasma resonance sensing signal, wherein the reference optical waveguide component has a first correction slope; a sensing optical waveguide component modified with a second noble metal nanoparticle layer further modified by an identification element, and another portion of incident light is introduced into the sensing optical waveguide component to generate a a second localized plasma resonance sensing signal, wherein the sensing optical waveguide component has a second correction slope; and a carrier that places the reference optical waveguide component and the sensing optical waveguide component; at least one light detecting unit that receives The first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal, wherein the first localized plasma resonance sensing signal comprises a blank solution generated by the reference optical waveguide component a first signal and a second signal generated by the reference optical waveguide component detecting a sample, the second localized plasma resonance sensing signal comprising a third generated by the sensing optical waveguide component detecting the blank solution a signal and a fourth signal generated by the sensing optical waveguide component detecting the sample; and a processor normalizing the first signal between the second signal and the first signal a difference and normalizing a second difference between the fourth signal and the third signal, wherein the processor uses the first correction slope and the second correction slope to adjust the second difference of normalization Obtaining an adjusted second difference, and then the processor utilizes a difference between the normalized first difference and the adjusted second difference to obtain a sensor response. The self-correcting localized plasma resonance sensing device of claim 12, wherein the first noble metal nanoparticle layer is modified on a reflective surface of the reference optical waveguide component. The self-correcting type localized plasma resonance sensing device of claim 12, wherein the second noble metal nanoparticle layer is reflected in the sensing optical waveguide component The surface is modified. The self-correcting type localized plasma resonance sensing device of claim 12, wherein the reference optical waveguide component and the sensing optical waveguide component are an optical fiber, a channel waveguide, a planar waveguide, or a tubular waveguide. The self-correcting type localized plasma resonance sensing device of claim 15, wherein the first noble metal nanoparticle layer is stripped to one of the optical fibers when the reference optical waveguide component is an optical fiber. Modifications are made at the area or at one end. The self-correcting type localized plasma resonance sensing device according to claim 15, wherein when the sensing optical waveguide component is an optical fiber, the second noble metal nanoparticle layer is stripped to one of the optical fibers. Modifications are made at the area or at one end. The self-correcting type localized plasma resonance sensing device according to claim 12, wherein the self-correcting type localized plasma resonance sensing device is a microfluidic wafer or a current sampling and analyzing device. The self-correcting type localized plasma resonance sensing device of claim 18, wherein the reference optical waveguide component and the sensing optical waveguide component are respectively at an end surface of the reference optical waveguide component and the sensing optical waveguide component One of the end faces constructs a mirror surface, wherein the reference optical waveguide component and the sense optical waveguide component are optical fibers or tubular waveguides. The self-correcting type localized plasma resonance sensing device of claim 19, wherein the reference optical waveguide component and the sensing optical waveguide component are further provided with a filter film and a rigid bracket having at least one opening, The mirror surface is configured to reflect the first localized plasma resonance sensing signal and the second localized plasma resonance sensing signal, and the filter filters an interference substance having a size larger than an average aperture of the filter, and The rigid support encases the reference optical waveguide component and the sensing optical waveguide component to increase the mechanical strength of the self-correcting localized plasma resonance sensing device during a sampling operation. The self-correcting localized plasma resonance sensing device according to claim 12, wherein the identification element comprises a chemical recognition molecule, an antibody, an antigen, a lectin, a hormone receptor, a nucleic acid or a saccharide. The self-correcting localized plasma resonance sensing device of claim 12, further comprising a lock-in amplifier having amplifying the first localized plasma resonance sensing signal and the second localized electric field The slurry resonance senses the signal and suppresses the function of the system noise. The self-correcting localized plasma resonance sensing device of claim 12, wherein the sensor response is represented by the following formula: Wherein ΔI _{S, SA} represents the sensor response caused by only specific adsorption, and I _S0 represents the third signal generated by the sensing optical waveguide component detecting the blank solution, and ΔI _S represents the fourth signal and the The second difference between the third signals, ΔI _{S, M} /I _S0 and ΔI _{R, M} /I _R0 respectively represent the normalized response of the sensing optical waveguide component and the reference optical waveguide component, which respectively represent The second correction slope and the first correction slope, ΔI _R represents the first difference between the second signal and the first signal, and I _R0 represents the detection of the blank solution by the reference optical waveguide component The first signal.