CN111551534A - Kit based on surface enhanced Raman probe, application thereof and imaging method - Google Patents
Kit based on surface enhanced Raman probe, application thereof and imaging method Download PDFInfo
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Abstract
The invention relates to a kit based on a surface enhanced Raman probe, and application and an imaging method thereof. The kit based on the surface enhanced Raman probe comprises a first surface enhanced Raman probe and a second surface enhanced Raman probe, wherein Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, and a targeting molecule is arranged on the surface of one of the first surface enhanced Raman probe and the second surface enhanced Raman probe. The kit based on the Raman probe, the application thereof and the imaging method can improve the positioning precision of the lymph node. The composite Raman probe has good enhancement performance and stability, does not have the problem of fluorescence quenching, and can be stored for a long time; the biocompatibility is good. In addition, because the Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, the specificity, the sensitivity and the accuracy of tumor cell imaging in the lymph node can be realized by detecting the abundance of the first surface enhanced Raman probe and the second surface enhanced Raman probe.
Description
Technical Field
The invention relates to the technical field of biological imaging, in particular to a kit based on a Raman probe, and application and an imaging method thereof.
Background
At present, cancer remains the first killer of human health, and the incidence rate is increasing year by year, and the treatment problem of cancer is still being tried to overcome worldwide. China is the country with the greatest increase in the rate of cancer growth worldwide. The postoperative metastasis spread of tumor surgical treatment is one of the main causes of cancer recurrence, and the prevention and treatment of tumor metastasis and recurrence is one of the important ways to improve the survival rate of cancer. The metastasis of tumor cells of primary cancer to the distant end through the lymph system via various levels of lymph nodes is the main mode of tumor metastasis. Therefore, in order to prevent tumor recurrence caused by tumor cell metastasis through lymphatic system, the solid tumor needs to be removed by surgery, and the lymphatic system which may have tumor cell metastasis needs to be cleared by lymphocleaning surgery, so as to achieve the purpose of reducing the recurrence risk.
However, the lymphatic system plays an important role in the vital physiological activities of the human body, and unnecessary cleaning can bring great influence to the mind and body of the patient, for example, breast cancer, and after axillary lymphatic cleaning, edema, dyskinesia, weakness and the like of the upper limbs can be caused, so that the life quality of the patient is seriously influenced. Therefore, accurate positioning of Sentinel Lymph nodes around a tumor and determination of whether the Lymph nodes have tumor metastasis are important bases for assessing whether to perform Lymph Node cleaning, if the Sentinel Lymph nodes have tumor cell metastasis, the risk of tumor metastasis through the lymphatic system is increased, Lymph cleaning is required, if the Sentinel Lymph nodes do not have tumor cell metastasis, the risk of tumor metastasis of a secondary lymphatic system is low, and Lymph cleaning can be avoided, so that the risk of tumor recurrence is reduced, the lymphatic system of a patient is protected, the survival and the life quality are improved, and introduction of the Sentinel Lymph Node Biopsy (SLNB) technology is called a revolution in breast cancer treatment history.
Sentinel Node (SLN) refers to the first Node to which a Lymph Node in the primary focus of a malignant tumor metastasizes. The metastatic status of regional lymph nodes can be judged by SLNB. Sentinel lymph node biopsy requires the use of a sentinel lymph node imaging agent and corresponding imaging techniques to accomplish the localization of sentinel lymph nodes, but existing imaging agents and imaging techniques suffer from certain drawbacks. At present, the sentinel lymph node imaging technology can only locate sentinel lymph nodes, but the accuracy is not high enough, and more importantly, the specific imaging of tumor cells in lymph nodes cannot be directly carried out.
At present, sentinel lymph node localization techniques widely used in clinical practice at home and abroad are first-generation and second-generation traditional techniques, the first-generation technique is a dyeing method, sentinel lymph nodes are dyed by carbon black or methylene blue, and are located by observing the dyed lymph nodes with naked eyes, but the reliability of the naked eye observation is poor, the retention time of a dyeing material in the first-station lymph nodes, namely the sentinel lymph nodes, is short, the dyeing material can quickly enter a secondary lymphatic system, the sentinel lymph nodes are difficult to accurately locate, and certain trouble is caused to doctors. The second generation technology is mainly used for positioning by a nuclide method and a fluorescent staining method, the nuclide method needs to detect signals sent by radioactive substances by means of external equipment to determine lymph nodes, certain potential safety hazards exist, and the nuclide method needs to be matched with nuclear medicine subjects, so that the nuclide method is inconvenient to use and difficult to popularize in large areas in clinic. The fluorescence method needs the help of fluorescence equipment, the price of the fluorescence equipment is high, and the fluorescence method has the problems of poor tissue penetrability, poor fluorescence stability, easy interference of tissue autofluorescence and the like. All the above techniques can only realize the positioning or imaging of sentinel lymph nodes, and cannot realize the specific imaging of tumor cells in lymph nodes.
Disclosure of Invention
Based on the above, it is necessary to provide a kit based on a surface enhanced raman probe, and an application and an imaging method thereof, aiming at the problems of how to improve positioning accuracy and imaging sensitivity.
A kit based on a surface enhanced Raman probe comprises a first surface enhanced Raman probe and a second surface enhanced Raman probe, wherein Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, and a targeting molecule is arranged on the surface of one of the first surface enhanced Raman probe and the second surface enhanced Raman probe.
In one embodiment, the first surface enhanced raman probe comprises a first metal core, a first raman reporter molecule coated on the first metal core, a first metal layer coated on the first raman reporter molecule; the second surface enhanced Raman probe comprises a second metal inner core, a second Raman reporter molecule coated on the second metal inner core, and a second metal layer coated on the second Raman reporter molecule;
alternatively, the first surface enhanced raman probe comprises a first metal core, a first raman reporter molecule coated on the first metal core, a first metal layer coated on the first raman reporter molecule, and a first protective layer coated on the first raman reporter molecule; the second surface enhanced Raman probe comprises a second metal inner core, a second Raman reporter molecule coated on the second metal inner core, a second metal layer coated on the second Raman reporter molecule, and a second protective layer coated on the second Raman reporter molecule.
In one embodiment, the first metal core and the second metal core are each selected from gold nanoparticles, silver nanoparticles, copper nanoparticles, aluminum nanoparticles, or platinum nanoparticles; the first Raman reporter molecule and the second Raman reporter molecule are respectively selected from one or more of dimercaptobenzene, 4-nitrothiophenol, o-nitrothiophenol, 4-toluene thiophenol, 2-mercapto-5-nitrobenzimidazole, 2-mercapto-6-nitrobenzothiazole, o-chlorothiophenol, 4-chlorobenzenethiol, biphenyl-4, 4' -dithiol and 2-naphthalene thiol; the first metal layer and the second metal layer are both selected from a gold layer, a silver layer, a copper layer, an aluminum layer or a platinum layer, or the first metal layer and the second metal layer are composite layers formed by sequentially laminating any two or more than two of the gold layer, the silver layer, the copper layer, the aluminum layer and the platinum layer; the first protective layer and the second protective layer are both selected from one or more of polymers and inorganic insulating materials.
In one embodiment, the targeting molecule is a specific targeting molecule.
In one embodiment, the targeting molecule is folate receptor or HER2 receptor.
In one embodiment, the paint also comprises a solvent.
In one embodiment, the composition further comprises a suspending agent, an acid-base regulator and/or an isotonic regulator.
The application of the kit based on the surface enhanced Raman probe in lymph node location and/or tumor cell imaging in lymph nodes is provided.
An imaging method of a kit based on a surface enhanced Raman probe comprises the following steps: allowing the kit to act on lymphocytes or tumor cells for a predetermined period of time; based on the kit, a Raman spectrometer is adopted for imaging to obtain a composite Raman spectrum.
In one embodiment, the method further comprises the following steps: decomposing the composite Raman spectrum to respectively obtain the abundance of the first surface enhanced Raman probe and the second surface enhanced Raman probe in the tumor cells; calculating a ratio value of the abundances of the first surface enhanced Raman probe and the second surface enhanced Raman probe in the lymphocyte or the tumor cell, and comparing the ratio value with a predetermined value.
Compared with the prior art, the invention has the beneficial effects that: the kit comprises a first surface enhanced Raman probe and a second surface enhanced Raman probe, wherein Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, one surface of the first surface enhanced Raman probe and one surface of the second surface enhanced Raman probe are provided with targeting molecules, lymph node can be rapidly imaged by detecting the Raman characteristic peaks and the strength of the first surface enhanced Raman probe and the second surface enhanced Raman probe, and in addition, the accuracy of lymph node positioning can be improved by adopting the composite surface enhanced Raman probe kit. The composite surface-enhanced Raman probe has good enhancement performance and good stability, does not have the problem of fluorescence quenching, and can be stored for a long time; the biocompatibility is good. In addition, because the Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, the specificity, the sensitivity and the accuracy of tumor cell imaging in the lymph node can be realized by detecting the abundance of the first surface enhanced Raman probe and the second surface enhanced Raman probe and calculating the abundance ratio.
Drawings
Fig. 1 is a schematic structural diagram of a first surface enhanced raman probe according to an embodiment;
FIG. 2 is a schematic structural diagram of a second surface enhanced Raman probe according to an embodiment;
FIG. 3 is a transmission electron micrograph of a first surface enhanced Raman probe of the present embodiment;
FIG. 4 is a transmission electron micrograph of a second surface enhanced Raman probe of the present embodiment;
FIG. 5 is an ultraviolet-visible absorption spectrum of the first surface enhanced Raman probe shown in FIG. 3;
FIG. 6 is an ultraviolet-visible absorption spectrum of the second surface enhanced Raman probe shown in FIG. 4;
FIG. 7 is a Raman spectrum of the first surface enhanced Raman probe shown in FIG. 3;
FIG. 8 is a Raman spectrum of the second surface enhanced Raman probe shown in FIG. 4;
FIG. 9 is a Raman spectrum of a composite surface enhanced Raman probe;
fig. 10 is a graph of the spectral signals of the raman spectrum shown in fig. 9 after decomposition.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Surface-Enhanced raman spectroscopy (SERS) refers to a very large enhancement effect of raman scattering of molecules adsorbed on the Surface of a metal nanostructure due to excitation of Surface plasmons and Surface chemical effects, and can realize single-molecule and single-particle detection, and has very high sensitivity. In addition, the unique fingerprint type map of the kit has ultrahigh specificity, the ultra-narrow half-width peak is beneficial to multi-index detection of optical coding, the optical stability is strong, and the problem of fluorescence quenching does not exist.
Based on the outstanding advantages, the SERS nanoprobe (sometimes also called surface enhanced Raman probe) has very wide application prospect in the biomedical field. Therefore, the novel sentinel lymph node positioning and imaging kit based on the Raman probe technology overcomes the defects of the prior art, and images sentinel lymph nodes of healthy people and cancer and tumor patients based on the surface enhanced Raman spectroscopy technology to improve the positioning accuracy and the imaging sensitivity of the lymph nodes. In addition, the method also helps to construct a new tumor screening and diagnosing method.
The kit based on the surface enhanced Raman probe of the embodiment comprises a first surface enhanced Raman probe and a second surface enhanced Raman probe, wherein Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, and a surface of one of the first surface enhanced Raman probe and the second surface enhanced Raman probe is provided with a targeting molecule. The range of the Raman signals of the first surface enhanced Raman probe and the second surface enhanced Raman probe is both 100-3000cm-1And the Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different. The surface of one of the first surface enhanced raman probe and the second surface enhanced raman probe has a targeting molecule, i.e. the surface of the first surface enhanced raman probe or the second surface enhanced raman probe has a targeting molecule. In this embodiment, the surface of the first surface enhanced raman probe is modified with a targeting molecule, and the surface of the second surface enhanced raman probe is not modified with the targeting molecule. In other embodiments, the surface of the second surface enhanced raman probe may be modified with a targeting molecule, and the surface of the first surface enhanced raman probe is not modified with the targeting molecule.
The kit comprises a first surface enhanced Raman probe and a second surface enhanced Raman probe, wherein Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, a targeting molecule is arranged on the surface of one of the first surface enhanced Raman probe and the second surface enhanced Raman probe, and lymph node can be rapidly imaged by detecting the Raman characteristic peaks and the strength of the first surface enhanced Raman probe and the second surface enhanced Raman probe. The composite surface-enhanced Raman probe has good enhancement performance and good stability, does not have the problem of fluorescence quenching, and can be stored for a long time; the biocompatibility is good. In addition, because the Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, the specificity, the sensitivity and the accuracy of tumor cell imaging in the lymph node can be realized by detecting the abundance of the first surface enhanced Raman probe and the second surface enhanced Raman probe and calculating the abundance ratio.
In one embodiment, the first surface enhanced raman probe includes a first metal core 110, a first raman reporter molecule 120 coated outside the first metal core 110, a first metal layer 130 coated outside the first raman reporter molecule 120, and a first protective layer 140 coated on the first metal layer 130. In addition, the surface of the first raman probe is modified with a targeting molecule 150, as shown in fig. 1.
Wherein the first metal core 110 is selected from gold nanoparticles, silver nanoparticles, copper nanoparticles, aluminum nanoparticles, or platinum nanoparticles. Preferably, the first metal core is a gold nanoparticle or a silver nanoparticle. Wherein the particle size of the first metal core may be 1 to 500 nm.
Wherein the first raman reporter molecule 110 is selected from one or more of dimercaptobenzene, 4-nitrothiophenol, o-nitrothiophenol, 4-toluene thiophenol, 2-mercapto-5-nitrobenzimidazole, 2-mercapto-6-nitrobenzothiazole, o-chlorothiophenol, 4-chlorothiophenol, biphenyl-4, 4' -dithiol and 2-naphthalene thiol. Further, the first raman reporter molecule 110 is selected from one of dimercaptobenzene, 4-nitrothiophenol, o-nitrothiophenol, 4-toluene thiophenol, 2-mercapto-5-nitrobenzimidazole, 2-mercapto-6-nitrobenzothiazole, o-chlorothiophenol, 4-chlorothiophenol, biphenyl-4, 4' -dithiol, and 2-naphthalene thiol.
Further, the first metal layer 130 may be a gold layer, a silver layer, a copper layer, an aluminum layer, or a platinum layer. Alternatively, the first metal layer may be formed by sequentially laminating any two or more of a gold layer, a silver layer, a copper layer, an aluminum layer, and a platinum layer. Preferably, the first metal layer is selected from a gold layer or a silver layer. Wherein, the thickness of the first metal layer can be 0.2-500 nm.
Further, the first protection layer 140 is selected from one or more of a polymer and an inorganic insulating material. Preferably, the first protective layer is selected from polymers such as polyamines, polyesters, polyols, and polyketones, such as polydopamine, polyaniline, polyethylene glycol, polyvinylpyrrolidone, and polydiacetylene, and the inorganic insulating material may be silicon dioxide, silicon nitride, and aluminum oxide. The thickness of the first protective layer 140 may be 0-300 nm. When the thickness of the first protective layer 140 is 0, that is, the first surface enhanced raman probe includes a first metal core 110, a first raman reporter 120 coated outside the first metal core 110, and a first metal layer 130 coated outside the first raman reporter 120. In this embodiment, the first metal core in the first surface-enhanced raman probe is gold nanoparticles, the first raman reporter molecule is 4-nitrothiophenol, the first metal layer is a gold layer, the first protective layer is a silicon dioxide layer, and the surface-modified targeting molecule is a folate receptor, as shown in fig. 3 and 5.
Similarly, the second surface enhanced raman probe includes a second metal core 210, a second raman reporter molecule 220 coated outside the second metal core 210, a second metal layer 230 coated outside the second raman reporter molecule 220, and a second protective layer 240 coated on the second metal layer 230, as shown in fig. 2. Wherein the second metal core 210 is selected from gold nanoparticles, silver nanoparticles, copper nanoparticles, aluminum nanoparticles, or platinum nanoparticles. Preferably, the second metal core 210 is a gold nanoparticle or a silver nanoparticle. Wherein the particle size of the second metal core 210 may be 1-500 nm.
Wherein the second raman reporter molecule 220 is selected from one or more of dimercaptobenzene, 4-nitrothiophenol, o-nitrothiophenol, 4-toluene thiophenol, 2-mercapto-5-nitrobenzimidazole, 2-mercapto-6-nitrobenzothiazole, o-chlorothiophenol, 4-chlorothiophenol, biphenyl-4, 4' -dithiol and 2-naphthalene thiol. Further, the second raman reporter molecule is selected from one of dimercaptobenzene, 4-nitrothiophenol, o-nitrothiophenol, 4-toluene thiophenol, 2-mercapto-5-nitrobenzimidazole, 2-mercapto-6-nitrobenzothiazole, o-chlorothiophenol, 4-chlorothiophenol, biphenyl-4, 4' -dithiol and 2-naphthalene thiol.
Further, the second metal layer 230 may be a gold layer, a silver layer, a copper layer, an aluminum layer, or a platinum layer. Alternatively, the second metal layer may be formed by sequentially laminating any two or more of a gold layer, a silver layer, a copper layer, an aluminum layer, and a platinum layer. Preferably, the second metal layer is a gold layer or a silver layer. Wherein the thickness of the second metal layer may be 0.2-500 nm.
Further, the second protective layer 240 is selected from one or more of a polymer, an inorganic insulating material. Preferably, the first protective layer is selected from polymers such as polyamine and polyester, the polyamine can be polydopamine or polyaniline, and the inorganic insulating material can be silicon dioxide, silicon nitride, aluminum oxide, and the like. Wherein, the thickness of the second protective layer can be 0-300 nm. In this embodiment, the second metal core in the second raman probe is gold nanoparticles, the second raman reporter molecule is dimercaptobenzene, the second metal layer is a gold layer, and the second protective layer is a silicon dioxide layer, as shown in fig. 4 and 6.
It should be noted that the first raman reporter molecule and the second raman reporter molecule are different, that is, the first raman reporter molecule and the second raman reporter molecule have different raman spectra, so that the first surface enhanced raman probe and the second surface enhanced raman probe have different raman characteristic peaks. As can be seen from fig. 5 and 6 and fig. 7 and 8, the first surface-enhanced raman probe and the second surface-enhanced raman probe have different extinction spectra (absorption intensities) and raman characteristic peaks. Therefore, when the kit is used for positioning the lymph nodes, the positioning precision is high.
In one embodiment, the targeting molecule is a specific targeting molecule. Further, the targeting molecule may be a folate receptor or a HER2 receptor. Further, the shape and structure of the first surface enhanced raman probe and the second surface enhanced raman probe may be nanospheres, nanorods, nanostars, and various core-shell structures and polymers, which are not limited herein.
In one embodiment, the kit further comprises a solvent, and the solvent can be physiological saline or the like.
The first surface-enhanced raman probe and the second surface-enhanced raman probe may be mixed by mechanical stirring, magnetic stirring, ultrasonic oscillation, vortex oscillation, or the like. In addition, the first surface-enhanced raman probe and the second surface-enhanced raman probe may be dispersed in a solution such as physiological saline. Further, the kit can also comprise a suspending agent, an acid-base regulator and/or an isotonic regulator.
The use process of the kit is as follows: the kit acts with lymphocytes or tumor cells, and after a period of time, the Raman spectrometer is adopted for detection and imaging to obtain a composite Raman spectrum. At this time, the obtained composite raman spectrum is a composite raman spectrum, and the composite raman spectrum is decomposed by algorithms such as a classical least square method, a nonnegative matrix decomposition method, a convolutional neural network method, a minimum absolute value convergence or a selective operator regression method, so as to obtain spectra of the first surface enhanced raman probe and the second surface enhanced raman probe and abundance in lymphocytes or tumor cells. The method for obtaining the abundance can be Raman single-point test, multipoint test, two-dimensional imaging or scanning, and three-position imaging or scanning. Accurate and specific imaging of tumor cells in sentinel lymph nodes is achieved by the ratio of the abundance of the two raman probes. If the ratio of the abundances of the two raman probes exceeds a predetermined ratio value, the presence of tumor cells is indicated. Wherein the lymphocyte can be lymph node tissue or lymph node extract. Further, the kit may be injected to a position where a lymph node is located in the body, or the kit may be injected around a solid tumor during an operation, and the kit may diffuse to a sentinel lymph node through a lymphatic system and stay under the action of lymphatic tropism.
The predetermined ratio value is related to the ratio of the first surface-enhanced raman probe to the second surface-enhanced raman probe, and if the ratio of the first surface-enhanced raman probe to the second surface-enhanced raman probe is 1:1 (intensity or area of the raman characteristic peak), the predetermined ratio value is also 1: 1. The ratio of the first surface-enhanced raman probe to the second surface-enhanced raman probe (the intensity or area of the raman characteristic peak) is not limited herein.
In this embodiment, as shown in fig. 9 and 10, the abundance of different signals such as the first raman probe, the second raman probe, the lymph node tissue, and the background can be obtained by classical least squares decomposition. And calculating the abundance value ratio of the first surface enhanced Raman probe and the second surface enhanced Raman probe.
The kit based on the surface enhanced Raman probe is applied to lymph node location and/or tumor cell imaging in lymph nodes. The kit based on the surface enhanced raman probe is described in the above embodiments, and is not described herein.
The kit is safe to use, has no problems of radioactivity and the like, and has high sentinel lymph node positioning accuracy. In addition, signals are obtained by means of a Raman device, and interference of human factors is avoided. The Raman probe has good enhancement performance and stability, has no problem of fluorescence quenching, and can be stored for a long time; the biocompatibility is good. Not only sentinel lymph node localization can be realized, but also whether tumor cells exist in sentinel lymph nodes can be judged.
An imaging method of the raman probe-based kit of an embodiment includes the steps of: allowing the kit to act on lymphocytes or tumor cells for a predetermined period of time; based on the kit, a Raman spectrometer is adopted for imaging to obtain a composite Raman spectrum. Wherein the kit can be co-incubated with lymphocytes. In this example, lymphocytes are taken as an example for explanation.
In one embodiment, the method further comprises the following steps: decomposing the detected Raman spectrum to respectively obtain the abundance of the first surface enhanced Raman probe and the second surface enhanced Raman probe in the lymphocyte or the tumor cell; and calculating a ratio value of the abundances of the first surface enhanced Raman probe and the second surface enhanced Raman probe in the lymphocyte or the tumor cell, and comparing the ratio value with a predetermined ratio value.
Specifically, the kit reacts with lymphocytes, and after a period of time, a Raman spectrometer is used for imaging to obtain a Raman spectrum. At this time, the obtained raman spectrum is a composite raman spectrum, and the composite raman spectrum is decomposed by algorithms such as a classical least square method, a nonnegative matrix decomposition method, a convolutional neural network method, a minimum absolute value convergence or a selective operator regression method, so as to obtain spectra of the first surface enhanced raman probe and the second surface enhanced raman probe and abundance in lymph node tissues or cells. The method for obtaining the abundance can be Raman single-point test, multipoint test, two-dimensional imaging or scanning, and three-position imaging or scanning. Accurate and specific imaging of tumor cells in sentinel lymph nodes is achieved by the ratio of the abundance of the two raman probes. If the ratio of the abundances of the two raman probes exceeds a predetermined ratio value, the presence of tumor cells is indicated. Wherein the lymphocyte can be lymph node tissue or lymph node extract. Further, the kit may be injected to a position where a lymph node is located in the body, or the kit may be injected around a solid tumor during an operation, and the kit may diffuse to a sentinel lymph node through a lymphatic system and stay under the action of lymphatic tropism. Therefore, the kit can realize the rapid diagnosis of the tumor metastasis condition of the sentinel lymph node in the operation.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A kit based on a surface enhanced Raman probe, comprising a first surface enhanced Raman probe and a second surface enhanced Raman probe, wherein Raman characteristic peaks of the first surface enhanced Raman probe and the second surface enhanced Raman probe are different, and a surface of one of the first surface enhanced Raman probe and the second surface enhanced Raman probe is provided with a targeting molecule.
2. The surface-enhanced Raman probe-based kit of claim 1, wherein the first surface-enhanced Raman probe comprises a first metal core, a first Raman reporter molecule coated on the first metal core, and a first metal layer coated on the first Raman reporter molecule; the second surface enhanced Raman probe comprises a second metal inner core, a second Raman reporter molecule coated on the second metal inner core, and a second metal layer coated on the second Raman reporter molecule;
or, the first surface enhanced raman probe comprises a first metal inner core, a first raman reporter molecule coated on the first metal inner core, a first metal layer coated on the first raman reporter molecule, and a first protective layer coated on the first metal layer; the second surface enhanced Raman probe comprises a second metal inner core, a second Raman reporter molecule coated on the second metal inner core, a second metal layer coated on the second Raman reporter molecule and a second protective layer coated on the second metal layer.
3. The surface-enhanced raman probe-based kit of claim 2, wherein the first metal core and the second metal core are each selected from gold nanoparticles, silver nanoparticles, copper nanoparticles, aluminum nanoparticles, or platinum nanoparticles; the first Raman reporter molecule and the second Raman reporter molecule are respectively selected from one or more of dimercaptobenzene, 4-nitrothiophenol, o-nitrothiophenol, 4-toluene thiophenol, 2-mercapto-5-nitrobenzimidazole, 2-mercapto-6-nitrobenzothiazole, o-chlorothiophenol, 4-chlorobenzenethiol, biphenyl-4, 4' -dithiol and 2-naphthalene thiol; the first metal layer and the second metal layer are both selected from a gold layer, a silver layer, a copper layer, an aluminum layer or a platinum layer, or the first metal layer and the second metal layer are composite layers formed by sequentially laminating any two or more than two of the gold layer, the silver layer, the copper layer, the aluminum layer and the platinum layer; the first protective layer and the second protective layer are both selected from one or more of polymers and inorganic insulating materials.
4. The surface-enhanced Raman probe-based kit of any one of claims 1-3, wherein said targeting molecule is a specific targeting molecule.
5. The surface-enhanced Raman probe-based kit of claim 4, wherein said targeting molecule is folate receptor or HER2 receptor.
6. The surface-enhanced Raman probe-based kit of any one of claims 1-3, further comprising a solvent.
7. The surface-enhanced Raman probe-based kit of claim 6, further comprising a suspending agent, an acid-base modifier, and/or an isotonicity modifier.
8. Use of a surface enhanced raman probe based kit according to any one of claims 1 to 7 for lymph node localization and/or imaging of tumor cells in lymph nodes.
9. An imaging method of a kit based on a surface enhanced Raman probe is characterized by comprising the following steps: allowing the kit to act on lymphocytes or tumor cells for a predetermined period of time; based on the kit, a Raman spectrometer is adopted for imaging to obtain a composite Raman spectrum.
10. The imaging method according to claim 9, further comprising the steps of: decomposing the composite Raman spectrum to respectively obtain the abundance of the first surface enhanced Raman probe and the second surface enhanced Raman probe in the lymphocyte or the tumor cell; calculating a ratio value of the abundances of the first surface enhanced Raman probe and the second surface enhanced Raman probe in the lymphocyte, and comparing the ratio value with a predetermined value.
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