CN212321444U - Detection device combining surface enhanced Raman scattering with SPR sensing - Google Patents
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Abstract
The utility model discloses a detection device combining surface enhanced Raman scattering with SPR sensing, which comprises an SPR detection unit and an SERS detection unit, wherein the SPR detection unit comprises an SPR excitation light source, an objective lens, a diaphragm, an incident lens, a prism, a micro-nano metal sensing chip, an emergent lens and a phase detector which are sequentially connected according to a light path, and the micro-nano metal sensing chip is positioned on the surface of the prism; the SERS detection unit comprises an SERS excitation light source, a collimator, a dichroic mirror, a micro-lens group A, an optical fiber, a micro-lens group B, a front objective, a Raman/fluorescence spectrometer and the micro-nano metal sensing chip. The invention improves the detection sensitivity and specificity, reduces the interference, has multiple channels, has high-flux application prospect, is convenient and quick to detect, has wide applicability and has multiple functions.
Description
Technical Field
The invention belongs to the field of optical sensing, and relates to a detection device combining surface enhanced Raman scattering with SPR sensing.
Background
For epidemic diseases caused by viruses, the current mainstream detection means is to detect nucleic acid by adopting a fluorescence quantitative PCR method, but still has the problems of long time consumption, complex operation, insufficient investigation sensitivity on early suspected patients and the like. Therefore, a new virus antibody detection technology which is more sensitive, rapid and efficient needs to be researched. Generally, the content of virus antibodies in the body fluid of a latent patient or a patient with weak immune function is low, and false negative is easily caused. How to improve the detection rate (reduce the occurrence of false negative) has important significance. Similarly, for the early detection of some tumors, the abundance of tumor antigens in blood is low, and high sensitivity is required for the detection of tumor antigens in blood. Sandwich methods for primary and secondary antibodies are generally used for tumor antigens, while indirect methods are generally used for detection of viral antibodies.
Surface Plasmon Resonance (SPR) is a physical optical phenomenon. When light is incident on the surfaces of the dielectric and the metal at a specific incident angle, the interface generates an SPR phenomenon, and the phase and intensity of reflected light are transited and attenuated. Since changes in the refractive index of the dielectric medium change the conditions under which the SPR phenomenon occurs, the optical properties of the emitted light respond very sensitively to changes in the refractive index. When the SPR phenomenon of the system is more pronounced, i.e. the intensity of the light is attenuated to a higher degree, the resonance state is more easily broken, i.e. the optical properties of the reflected light are more sensitive to the change of the refractive index.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a detection device combining surface enhanced Raman scattering and SPR sensing.
A detection device combining surface enhanced Raman scattering with SPR sensing comprises an SPR detection unit and an SERS detection unit, wherein the SPR detection unit comprises an SPR excitation light source, an objective lens, a diaphragm, an incident lens, a prism, a micro-nano metal sensing chip, an exit lens and a phase detector which are sequentially connected according to a light path, and the micro-nano metal sensing chip is positioned on the surface of the prism;
the SERS detection unit comprises an SERS excitation light source, a collimator, a dichroic mirror, a micro-lens group A, an optical fiber, a micro-lens group B, a front objective, a Raman/fluorescence spectrometer and the micro-nano metal sensing chip;
laser emitted by the SERS excitation light source irradiates to a dichroic mirror after passing through a collimator, is coupled to an optical fiber after being reflected through a micro lens group A, and is emitted through a micro lens group B and then enters a micro-nano metal sensing chip, the micro lens group A, the optical fiber and the micro lens group B form a multi-channel optical fiber probe array, signals excited by the micro-nano metal sensing chip are received by the multi-channel optical fiber probe array again and then are emitted from the dichroic mirror after being collimated, and then the signals are collected by a front object placing mirror of a Raman/fluorescence spectrum instrument and then are transmitted to the Raman/fluorescence spectrum instrument.
Furthermore, a layer of medium film with a periodic micro-nano empty groove array is paved on the front surface of the micro-nano metal sensing chip, the size of an opening of the empty groove is suitable for preventing large-size non-specific substances from passing through the empty groove, non-specific adsorption is reduced, and accurate measurement of the number of Raman/fluorescence probes in direct proportion to the concentration of the substances to be measured is facilitated.
The invention has the beneficial effects that:
the kit has the advantages of improving detection sensitivity and specificity, reducing interference, having multiple channels, having a high-flux application prospect, being convenient and quick to detect, being wide in applicability and having multiple functions.
Drawings
FIG. 1 is a schematic structural diagram of a detection apparatus combining surface enhanced Raman scattering and SPR sensing in example 1 of the present invention;
description of reference numerals: the device comprises an SPR excitation light source 1, an objective lens 2, a diaphragm 3, an incident lens 4, a prism 5, a micro-nano metal sensing chip 6, an emergent lens 7, a phase detector 8, an SERS excitation light source 9, a collimator 10, a dichroic mirror 11, a micro-lens group A12, an optical fiber 13, a micro-lens group B14, a front objective lens 15 and a Raman/fluorescence spectrometer 16.
Detailed Description
The invention is further illustrated by the following principles and examples.
Basic principle
The metal material with the sub-wavelength structure has the advantage of adjustable refractive index, and the micro-nano structure sensing film with strong SPR phenomenon can be designed through the electromagnetic theory, so that the resolution of the SPR sensor is improved.
On one hand, the detection of the tiny refractive index change is realized by constructing a high-sensitivity phase SPR sensor, so that the detection sensing is carried out on the tiny refractive index change generated after the detection of the combination of the tumor antigen and the corresponding antibody on the sensing surface or the combination of the IgG and IgM proteins (primary antibodies) of the virus and the corresponding antigen on the sensing surface, and the high sensitivity is realized. The phase-type SPR has high sensitivity, so that the detection rate of target protein can be increased. There are many substances to be tested for, and the above tumor antigens or antibodies to viruses are only used for exemplary purposes.
On the other hand, the raman spectrum in scattered light is related to vibration and rotation inside scattering molecules, which have fingerprint properties. Raman scattering of water is weak, so raman spectroscopy is an ideal means for studying biological samples and the like in aqueous solutions. The spectrum peak of the Raman spectrum is clear and sharp, and the method is suitable for quantitative research. In chemical structure analysis, the intensity of the independent raman intervals can be correlated with the number of functional groups. The accuracy of the raman spectrum is therefore higher relative to the fluorescence spectrum. However, raman spectra are generally weak, and a metal micro-nano structure engraved with a special pattern can realize a signal of Surface Enhanced Raman Scattering (SERS). An excitation light source of a Raman/fluorescence spectrum system can irradiate the front side of a sensing chip with a special metal micro-nano structure, if a tumor antigen is detected, the front side of the sensing chip is connected with a primary antibody protein corresponding to a target protein, the target protein can be captured, a secondary antibody protein correspondingly modified with a Raman probe can be connected to the surface of the target protein, and a Surface Enhanced Raman Spectrum (SERS) signal can be obtained by the metal micro-nano structure with a special pattern. The SERS reflection light path on the front side of the sensing chip collects Raman spectrum signals generated by the Raman probe through the micro-lens group and the optical fiber group, and a spatial distribution diagram of the Raman signals is read by the Raman spectrometer, so that the detection accuracy of the system is further improved.
A detection method combining surface enhanced Raman with SPR sensing is characterized in that an SPR excitation light source irradiates the back surface of a micro-nano metal sensing chip, a prism excites the back surface of the micro-nano metal sensing chip to generate an SPR resonance phenomenon, and a phase detection device reads a signal (SPR signal) of refractive index change near the front surface of the micro-nano metal sensing chip carried by a reflected light phase;
the SERS excitation light source irradiates the front side of the micro-nano metal sensing chip, Raman spectrum signals are collected and collected through the micro-lens group and the optical fiber group, and Raman scattering signals (SERS signals) are read by the Raman signal detection device;
the micro-nano metal sensing chip simultaneously supports an SPR guided wave mode and an SERS signal for enhancing Raman scattering/fluorescence;
the front surface of the micro-nano metal sensing chip is provided with a first bonding material of a substance to be detected, after the substance to be detected is specifically bonded with the first bonding material, a second bonding material of the substance to be detected is connected with the first bonding material through the substance to be detected, and the second bonding material of the substance to be detected is provided with a Raman probe or a fluorescent probe; the SPR signal reflects the refractive index change caused by the concentration of the substance to be detected, and the concentration of the substance to be detected is positively correlated with the SERS signal.
The Raman signal detection device is a Raman spectrum instrument, namely a spectrum instrument with high spectral resolution, and is suitable for Raman scattering and fluorescence spectra.
The Raman signal detection device is a Raman spectrometer, a filter plate is arranged to filter out light with characteristic wavelength, a two-dimensional detection camera of the spectrometer performs imaging once to avoid space scanning, and the spatial distribution of the surface enhanced fluorescence signal on the micro-nano metal sensing chip is detected.
Furthermore, a layer of medium film with a periodic micro-nano empty groove array is paved on the front surface of the micro-nano metal sensing chip, the size of an opening of the empty groove is suitable for preventing large-size non-specific substances from passing through the empty groove, non-specific adsorption is reduced, and accurate measurement of the number of Raman/fluorescence probes in direct proportion to the concentration of the substances to be measured is facilitated.
The invention is further elucidated with reference to the figures and embodiments.
Example 1
The embodiment discloses a detection device combining surface enhanced Raman scattering and SPR sensing, which comprises an SPR detection unit and an SERS detection unit.
The SPR detection unit comprises an SPR excitation light source 1, an objective lens 2, a diaphragm 3, an incident lens 4, a prism 5, a micro-nano metal sensing chip 6, an exit lens 7 and a phase detector 8 which are sequentially connected according to a light path, wherein the micro-nano metal sensing chip 6 is positioned on the surface of the prism 5.
The SERS detection unit comprises an SERS excitation light source 9, a collimator 10, a dichroic mirror 11, a micro-lens group A12, an optical fiber 13, a micro-lens group B14, a front objective 15, a Raman/fluorescence spectrum analyzer 16 and the micro-nano metal sensing chip 6.
The SPR excitation light source 1 emits light, the light is collimated and expanded through the objective lens 2, the diaphragm 3 and the incident lens 4, and then the prism 5 excites the micro-nano metal sensing chip 6 to generate an SPR phenomenon. According to the principle of the SPR phenomenon, the more sensitive the phase change of the reflected light from the micro-nano metal sensing chip 6 is to the change of the refractive index near the front surface of the micro-nano metal sensing chip 6, the more obvious the SPR phenomenon is, the higher the sensitivity of the phase change of the reflected light to the change of the refractive index is. In the embodiment, the metal engraved with the special micro-nano pattern is used as the sensing film material, and a high-sensitivity sensing film pattern can be designed through a simulation method. Taking the detection of tumor antigens as an example, the surface of the micro-nano metal sensing chip 6 is connected with a primary antibody protein of target protein, when the target protein exists in the object to be detected, the primary antibody protein captures the target protein, so that the equivalent refractive index of the surface of the micro-nano metal sensing chip 6 is changed, and the phase of reflected light detected by the phase detector 8 is obviously changed. And injecting a second antibody protein which is modified with a Raman probe and corresponds to the target protein into the micro-nano metal sensing chip 6, wherein the second antibody protein is combined with the target protein captured on the surface of the sensing membrane.
The SERS detection unit can perform synchronous spectrum detection on a plurality of channels of the micro-fluidic substrate of the micro-nano metal sensing chip 6, and the detection precision of target protein is improved by detecting enhanced Raman fluorescent signals. In this embodiment, the detection device further adopts an embodiment of enhancing raman scattering/fluorescence signal to improve detection accuracy by taking 4 detection channels as an example.
In this embodiment, the SERS excitation light source 9 adopts a single point laser, and a laser beam is corresponding to a channel on each micro-nano metal sensor chip 6. Laser irradiates to a dichroic mirror 11 after passing through a collimator 10, is coupled into an optical fiber 13 after being reflected through a micro lens group A12, is emitted through a micro lens group B14, the micro lens group A12, the optical fiber and the micro lens group B14 form a multi-channel optical fiber probe array, and finally is emitted from the micro lens group B14 and then is incident on a channel of a micro nano metal sensing chip 6 to excite a Raman probe modified on the anti-albumin to emit Raman or fluorescence signals. The excited signal is received by the multi-channel fiber probe array again, collimated and emitted from the dichroic mirror 11, and then received by the front objective 15 of the raman/fluorescence spectrometer 16. The front objective 15 of the raman/fluorescence spectrometer 16 performs line area imaging on a plurality of channels of the sensor chip, and an image plane is located at a slit of the spectrometer. The raman/fluorescence spectrometer 16 uses an area-array camera and is capable of detecting spectral images of line regions after being split by a grating. The multi-channel fiber probe array is conjugated with the slit, and compared with a line laser excitation signal, the multi-channel fiber probe array can effectively improve the excitation efficiency and is convenient to adjust, and the synchronism of signal detection is ensured by using a graph spectrometer of an area array detector, and each channel is simultaneously acquired. The fiber optic probe can also be changed to a spatial light input according to specific requirements.
According to the description, the metal surface micro-nano structure of the micro-nano metal sensing chip is designed to play a role in supporting an SPR guided wave mode and a Surface Enhanced Raman Scattering (SERS) signal simultaneously, so that the sensitivity of target protein detection is improved; in addition, the SERS detection unit is introduced to the other side of the sensing chip, and the robustness of noise resistance is obtained by analyzing the collected multi-mode sensing data through a computer, so that the detection resolution is further improved. The invention has the characteristics of high detection sensitivity and high resolution.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (2)
1. A detection device combining surface enhanced Raman scattering and SPR sensing is characterized in that,
the device comprises an SPR detection unit and an SERS detection unit, wherein the SPR detection unit comprises an SPR excitation light source, an objective lens, a diaphragm, an incident lens, a prism, a micro-nano metal sensing chip, an exit lens and a phase detector which are sequentially connected according to a light path, wherein the micro-nano metal sensing chip is positioned on the surface of the prism;
the SERS detection unit comprises an SERS excitation light source, a collimator, a dichroic mirror, a micro-lens group A, an optical fiber, a micro-lens group B, a front objective, a Raman/fluorescence spectrometer and the micro-nano metal sensing chip;
laser emitted by the SERS excitation light source irradiates to a dichroic mirror after passing through a collimator, is coupled to an optical fiber after being reflected through a micro lens group A, and is emitted through a micro lens group B and then enters a micro-nano metal sensing chip, the micro lens group A, the optical fiber and the micro lens group B form a multi-channel optical fiber probe array, signals excited by the micro-nano metal sensing chip are received by the multi-channel optical fiber probe array again and then are emitted from the dichroic mirror after being collimated, and then the signals are collected by a front object placing mirror of a Raman/fluorescence spectrum instrument and then are transmitted to the Raman/fluorescence spectrum instrument.
2. The detection device according to claim 1, wherein a dielectric film with a periodic micro-nano empty groove array is further laid on the front surface of the micro-nano metal sensing chip, and the size of an opening of the empty groove is suitable for preventing large-size non-specific substances from passing through the empty groove.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111208114A (en) * | 2020-03-07 | 2020-05-29 | 浙江大学 | Detection method and device for surface enhanced Raman scattering/fluorescence combined SPR sensing |
| CN114813697A (en) * | 2022-03-16 | 2022-07-29 | 江苏暖阳半导体科技有限公司 | High-flux multi-channel rapid Raman detection system of micro LED chip |
| US20230168239A1 (en) * | 2021-11-27 | 2023-06-01 | M.I.S. Electronics Inc. | Multimodal nanobiosensor for detecting cancer biomarkers in blood serum |
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2020
- 2020-03-07 CN CN202020270334.1U patent/CN212321444U/en active Active
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111208114A (en) * | 2020-03-07 | 2020-05-29 | 浙江大学 | Detection method and device for surface enhanced Raman scattering/fluorescence combined SPR sensing |
| US20230168239A1 (en) * | 2021-11-27 | 2023-06-01 | M.I.S. Electronics Inc. | Multimodal nanobiosensor for detecting cancer biomarkers in blood serum |
| US11828747B2 (en) * | 2021-11-27 | 2023-11-28 | M.I.S. Electronics Inc. | Multimodal nanobiosensor for detecting cancer biomarkers in blood serum |
| CN114813697A (en) * | 2022-03-16 | 2022-07-29 | 江苏暖阳半导体科技有限公司 | High-flux multi-channel rapid Raman detection system of micro LED chip |
| CN114813697B (en) * | 2022-03-16 | 2022-12-27 | 江苏暖阳半导体科技有限公司 | High-flux multi-channel rapid Raman detection system of micro LED chip |
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