CN115468944A - Monomolecular Raman fiber optical tweezers based on core-shell microlens and manufacturing method thereof - Google Patents
Monomolecular Raman fiber optical tweezers based on core-shell microlens and manufacturing method thereof Download PDFInfo
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Abstract
The invention discloses monomolecular Raman fiber optical tweezers based on a core-shell microlens and a manufacturing method thereof, wherein the monomolecular Raman fiber optical tweezers comprise: the device comprises a Raman spectrometer, an optical fiber coupler, an optical fiber laser, an optical fiber probe and a core-shell micro-lens suspension liquid for containing a sample to be detected, wherein the front end of the optical fiber probe extends into the core-shell micro-lens suspension liquid. The embodiment of the invention can amplify the Raman signal of the sample by using the echo wall resonance effect of the core-shell micro lens, thereby realizing the detection of the nanoscale biological sample in the biological environment, and the invention can complete the detection without introducing exogenous substances and has high biological compatibility; based on the miniaturization of the core-shell microlens fiber probe, the single-molecule Raman spectrum can be detected in a narrow biological environment; the invention has the capability of capturing a single biomolecule, can amplify a Raman scattering signal of the biomolecule, and can be used for in-situ detection of biomacromolecules or nano-scale bacteria or viruses in a biological environment.
Description
Technical Field
The invention relates to the field of biomedical detection, in particular to monomolecular Raman fiber optical tweezers based on a core-shell microlens and a manufacturing method thereof.
Background
Raman microscopy is widely used in the field of biomedical detection, and its bulky instrument is difficult to be applied to narrow biological environments such as blood vessels, intestines, esophagus and the like. In order to accurately realize the accuracy of Raman spectrum detection of biological macromolecules and enable the detection accuracy of Raman spectrum of the biological macromolecules to reach the level of single molecules, three problems exist in the technology: firstly, the size of the biomacromolecule is in the nanometer scale, so the diameter of the Raman scattering light spot also needs to be in the nanometer scale, and the accurate detection of the single biomacromolecule can be realized. Secondly, a small and flexible optical probe capable of penetrating into a living body environment is needed to realize the detection of biomacromolecules in the living body environment. Finally, the Raman scattering signals are usually weak due to the size of the biological macromolecules, so that the Raman scattering signals of the target object are difficult to effectively collect in a complex biological environment,
disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides the monomolecular Raman fiber optical tweezers based on the core-shell micro lens and the manufacturing method thereof, the structure is simple, the probe is small and exquisite and flexible, a biomacromolecule Raman scattering signal can be amplified at a detection position, and the detection of a nanoscale biological sample in a biological environment is realized.
According to the embodiment of the first aspect of the invention, the monomolecular Raman fiber optical tweezers based on the core-shell micro lens comprises: a Raman spectrometer; the Raman spectrometer is connected with a first end of the optical fiber coupler; the optical fiber laser is connected with the second end of the optical fiber coupler through an isolator; the third end of the optical fiber coupler is connected with the interface end of the optical fiber probe; the optical fiber probe comprises a core-shell micro-lens suspension liquid used for containing a sample to be detected, and the front end of the optical fiber probe extends into the core-shell micro-lens suspension liquid.
According to the first aspect of the invention, the monomolecular Raman fiber optical tweezers based on the core-shell microlens have at least the following beneficial effects:
the embodiment of the invention can amplify the Raman signal of the sample by using the echo wall resonance effect of the core-shell micro lens, thereby realizing the detection of the nanoscale biological sample in a biological environment, can complete the detection without introducing exogenous substances, and has high biological compatibility; based on the miniaturization of the core-shell microlens fiber probe, the single-molecule Raman spectrum can be detected in a narrow biological environment; the invention has the capability of capturing a single biomolecule, can amplify a Raman scattering signal of the biomolecule, and can be used for in-situ detection of biomacromolecules or nano-scale bacteria or viruses in a biological environment.
According to some embodiments of the invention, the fiber laser emits laser light at a wavelength of 785nm.
According to some embodiments of the invention, the core-shell microlens suspension is a TiO2/SiO2 core-shell composite particle suspension.
According to some embodiments of the invention, the diameter of the optical fiber probe is 3 to 5 μm, and the taper angle of the optical fiber probe is 60 to 73 °.
According to the embodiment of the second aspect of the invention, the method for manufacturing the monomolecular Raman fiber optical tweezers based on the core-shell micro lens comprises the following steps:
the method comprises the steps of obtaining an optical fiber probe, connecting the optical fiber probe to a third end of an optical fiber coupler, connecting a Raman spectrometer to a first end of the optical fiber coupler, and connecting an optical fiber laser to a second end of the optical fiber coupler through an isolator;
preparing a core-shell microlens suspension;
and dripping the suspension of the core-shell micro-lens onto a glass slide, and extending the front end of the optical fiber probe into the suspension of the core-shell micro-lens.
According to the embodiment of the second aspect of the invention, the method for manufacturing the monomolecular Raman fiber optical tweezers based on the core-shell microlens at least has the following beneficial effects:
the embodiment of the invention can amplify the Raman signal of the sample by using the echo wall resonance effect of the core-shell micro lens, thereby realizing the detection of the nanoscale biological sample in a biological environment, can complete the detection without introducing exogenous substances, and has high biological compatibility; based on the miniaturization of the core-shell microlens fiber probe, the single-molecule Raman spectrum can be detected in a narrow biological environment; the invention has the capability of capturing a single biomolecule, can amplify a Raman scattering signal of the biomolecule, and can be used for in-situ detection of biomacromolecules or nano-scale bacteria or viruses in a biological environment.
According to some embodiments of the invention, the method for manufacturing the optical fiber probe comprises
Cutting the optical fiber into two sections from the middle, stripping off the plastic outer skin and the coating layer of the middle section of the optical fiber to obtain a section of bare optical fiber, and sleeving the optical fiber into the metal thin tube;
the bare optical fiber is melted by a carbon dioxide laser, and then the melted portion is thinned, thereby drawing the optical fiber probe.
According to some embodiments of the present invention, the specific steps for preparing the core-shell microlens suspension are as follows:
dispersing a titanium precursor in absolute ethyl alcohol to obtain a titanium precursor solution;
adding SiO2 microsphere powder into absolute ethyl alcohol to obtain an ethyl alcohol dispersion liquid of SiO2 microspheres;
stirring the titanium precursor solution, dropwise adding the titanium precursor solution into the ethanol dispersion liquid of the SiO2 microspheres, and then putting the titanium precursor solution into a constant-temperature reaction container for reaction to obtain core-shell particle ethanol dispersion liquid;
and taking out the core-shell particle ethanol dispersion liquid and diluting the core-shell particle ethanol dispersion liquid by deionized water to obtain the core-shell micro-lens suspension liquid.
According to some embodiments of the invention, the reaction temperature in the reaction vessel is between 28 ℃ and 42 ℃.
According to some embodiments of the invention, the SiO2 microsphere powder has a diameter of 3 to 5 μm.
According to some embodiments of the invention, the thickness of the core-shell in the core-shell particle ethanol dispersion is 0.05 to 0.1 μm.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The invention is further described with reference to the following figures and examples, in which:
FIG. 1 is a schematic diagram of a principle of monomolecular Raman fiber optical tweezers based on a core-shell microlens in an embodiment of the present invention;
FIG. 2 is a graph of the intensity distribution of a light field in an embodiment of the present invention;
FIG. 3 is a graph showing the intensity of a photon nano jet light field generated by a core-shell microlens in an embodiment of the present invention.
Reference numbers:
the Raman spectrometer comprises a Raman spectrometer 100, an optical fiber coupler 200, an optical fiber laser 300, an optical fiber probe 400, a core-shell micro-lens suspension 500, a core-shell micro-lens 510, an isolator 600, an optical fiber adjusting frame 700, a glass slide 800 and a sample to be detected 900.
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
In the description of the present invention, it should be understood that the orientation descriptions, such as the orientation or positional relationship indicated by upper, lower, etc., are based on the orientation or positional relationship shown in the drawings, and are only for convenience of description and simplification of the description, but do not indicate or imply that the device or element referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus should not be construed as limiting the present invention.
In the description of the present invention, a plurality means two or more. If there is a description of first and second for the purpose of distinguishing technical features only, this is not to be understood as indicating or implying a relative importance or implicitly indicating the number of technical features indicated or implicitly indicating the precedence of technical features indicated.
In the description of the present invention, unless otherwise explicitly limited, terms such as arrangement, installation, connection and the like should be understood in a broad sense, and those skilled in the art can reasonably determine the specific meanings of the above terms in the present invention in combination with the specific contents of the technical solutions.
Referring to fig. 1, a monomolecular raman fiber optical tweezers based on a core-shell microlens includes: raman spectrometer 100, fiber coupler 200, fiber laser 300, fiber probe 400, and core-shell microlens suspension 500. In this embodiment, the optical fiber coupler 200 is a Y-shaped optical fiber coupler 200, the raman spectrometer 100 is connected to one arm at the left end of the optical fiber coupler 200, the optical fiber laser 300 is connected to the other arm at the left end of the optical fiber coupler 200 through an isolator 600, the right end of the optical fiber coupler 200 is connected to the interface end of the optical fiber probe 400, the optical fiber probe 400 is fixed on the optical fiber adjusting frame 700, the core-shell microlens suspension 500 containing the sample 900 to be measured is placed on the glass slide 800, and the front end of the optical fiber probe 400 extends into the core-shell microlens suspension 500.
The fiber laser 300 is used for emitting the trapping light and the excitation light, and the wavelength of the fiber laser 300 is 785nm and the power of the fiber laser 300 is 10-60 mW in this embodiment, but the fiber laser 300 with other wavelengths and powers may be selected.
The core-shell microlens suspension 500 is a TiO2/SiO2 core-shell composite particle suspension, and a core-shell microlens 510 suspension made of other microcavity materials can be adopted, such as silicon compounds, titanium dioxide, crystals, polymers and the like, because the surface of the optical fiber can generate electrostatic adsorption force, the core-shell microlens 510 in the core-shell microlens suspension 500 can be adhered to the tip of the optical fiber probe 400, the core-shell microlens 510 optical fiber probe is obtained, an echo wall effect can be generated under laser with a specific wavelength, and a Raman scattering signal of a molecule can be amplified. In this embodiment, the diameter of the optical fiber probe 400 is 3 to 5 μm, and the taper angle of the optical fiber probe 400 is 60 to 73 °.
The echo wall resonance effect of the core-shell micro lens 510 is utilized to amplify the Raman signal of the sample, so that the nano-scale biological sample can be detected in the biological environment, and the detection can be completed without introducing exogenous substances, so that the biological sample has high biocompatibility; based on the miniaturization of the core-shell micro-lens 510 optical fiber probe, the single-molecule Raman spectrum can be detected in a narrow biological environment; the invention has the capability of capturing a single biomolecule, can amplify a Raman scattering signal of the molecule, and can be used for in-situ detection of biomacromolecules or nano-scale bacteria or viruses in a biological environment.
The invention also relates to a manufacturing method of the monomolecular Raman fiber optical tweezers based on the core-shell micro lens, which comprises the following steps:
s100, obtaining the optical fiber probe 400, connecting the optical fiber probe 400 to the third end of the optical fiber coupler 200, connecting the Raman spectrometer 100 to the first end of the optical fiber coupler 200, and connecting the optical fiber laser 300 to the second end of the optical fiber coupler 200 through the isolator 600.
It should be noted that, in the embodiment of the present invention, the optical fiber probe 400 is manufactured by a fusion tapering method through a large-core optical fiber, and the optical fiber is first cut into two sections from the middle, and a plastic outer skin and a coating layer of the middle section of the optical fiber are stripped off to obtain a section of bare optical fiber, and then the optical fiber is sleeved in a metal thin tube; the bare optical fiber is melted by a carbon dioxide laser, and then the melted portion is thinned, thereby drawing the optical fiber probe 400.
It should be noted that the specific steps of the large-core optical fiber by the fusion tapering method are as follows:
s101, selecting the type of the optical fiber, wherein the optical fiber in the embodiment selects a large-core optical fiber, the core diameter of the large-core optical fiber is 100 mu m, and the type of the connector is FC/PC.
S102, stripping a coating layer at the middle section of the large-core-diameter optical fiber by using an optical fiber wire stripper to obtain a section of bare large-core-diameter optical fiber, sleeving the large-core-diameter optical fiber in a metal thin tube, and protecting the large-core-diameter optical fiber through the metal thin tube, wherein the inner diameter of the metal thin tube is 0.9-1.0 mm, the wall thickness is 0.08-0.12 mm, and the length is 10-12 cm in the embodiment.
S103, horizontally placing the bare large-core optical fiber above a carbon dioxide laser, standing for several minutes to heat the optical fiber to the temperature of about 500 ℃ so as to melt the large-core optical fiber, then thinning the melted part at the speed of 3-5 mm/S, and drawing into the optical fiber with the diameter of 3-5 microns, the length of 10 microns and the cone angle of 60-73 degrees, thus obtaining the required optical fiber probe 400.
S200, preparing a core-shell microlens suspension 500, taking TiO2/SiO2 core-shell composite particle suspension as an example, and specifically comprising the following steps:
s201, dispersing a titanium precursor in absolute ethyl alcohol to obtain a titanium precursor solution; wherein the titanium precursor is titanium alkoxide (TBT), and the concentration of the titanium precursor solution is about 0.01mol/L.
S202, adding the SiO2 microsphere powder into absolute ethyl alcohol to obtain an ethyl alcohol dispersion liquid of the SiO2 microspheres; wherein the diameter of the SiO2 microsphere powder is 3-5 μm.
S203, dropwise adding the titanium precursor solution prepared in the step S201 into the ethanol dispersion liquid of the SiO2 microspheres prepared in the step S202 while stirring by using a constant-temperature stirrer, then putting into a constant-temperature reaction container, wherein the reaction temperature in the reaction container is 28-42 ℃, the reaction is carried out for about 24 hours at 30 ℃ in the embodiment, carrying out appearance characterization on the prepared microlens through a transmission electron microscope, ensuring that the thickness of the core-shell structure is within a specified range, and controlling the thickness of the TiO2 shell to be 0.05-0.1 mu m, thereby obtaining the core-shell particle ethanol dispersion liquid.
And S204, taking out the core-shell particle ethanol dispersion liquid, diluting the core-shell particle ethanol dispersion liquid by using deionized water, and then diluting the core-shell particle ethanol dispersion liquid by using the deionized water until the concentration is 4.0 multiplied by 104 core-shell microlenses 510 per microliter to obtain the core-shell microlens suspension liquid 500.
It should be noted that each single particle in the core-shell microlens suspension 500 can be regarded as a microlens having a core-shell structure, the refractive index of the core and the refractive index of the shell are 2-2.5 and 1.4-1.5, respectively, and the microlens in the present invention functions as an echo wall microcavity coupled with a tapered optical fiber to generate an echo wall effect, and it should be understood that, besides TiO2, materials for manufacturing such a microcavity include silicon compounds, titanium dioxide, crystals, polymers, and the like.
S300, placing a glass slide 800 on a stage, dripping the core-shell micro-lens suspension 500 on the glass slide, and extending the front end of the optical fiber probe 400 into the core-shell micro-lens 510 solution to obtain the core-shell micro-lens 510 optical fiber probe.
The monomolecular Raman fiber optical tweezers based on the core-shell microlens can be obtained through the steps from S100 to S300, and the specific working procedures are described as follows:
the fiber laser 300 is opened, laser is introduced into the fiber probe 400 through the fiber coupler 200, an optical gradient force is generated at the front end of the fiber probe 400, the core-shell microlens 510 in the core-shell microlens suspension 500 is captured, the core-shell microlens 510 is adhered to the tip of the fiber probe due to the electrostatic adsorption force generated on the surface of the fiber, the light with a specific wavelength is coupled by using an evanescent field and a microsphere due to the total reflection principle of the light, the light resonates in a microcavity to form a stable standing wave, namely, an echo wall effect is generated, and meanwhile, a light potential well is generated at the front end of the core-shell microlens 510 due to the fact that the focus of the tapered fiber is larger than the diameter of the microlens, and the core-shell microlens 510 fiber probe is obtained. Other microlenses such as biological microlenses can also produce echo wall effect under laser with specific wavelength under the premise of meeting the requirements of shape, size and refractive index.
When raman spectrum probing is performed, a prepared sample 900 to be tested is placed on a glass slide 800, a prepared core-shell microlens 510 fiber probe is extended into the sample 900 to be tested, the fiber laser 300 is opened to emit laser with a wavelength of 785nm, the laser reaches the captured sample 900 to be tested through the isolator 600, the fiber coupler 200, the fiber probe 400 and the core-shell microlens 510 in sequence, a raman scattering signal of the sample 900 to be tested is excited, and referring to fig. 2, it can be known that 796nm laser generates an echo wall effect after passing through the tapered fiber with the shape and the core-shell microlens 510 with a diameter of 3 μm, and an optical potential well is formed at the front end of the core-shell microlens 510. The raman scattering signal is enhanced by the echo wall resonance effect of the core-shell microlens 510 and then transmitted to the raman spectrometer 100 through the fiber probe 400 and the fiber coupler 200.
Referring to fig. 3, it is shown that, as a graph of the optical field intensity of the photon nanometer jet generated by the core-shell microlens 510, FWHM =0.23 λ breaks through the diffraction limit, so that the core-shell microlens 510 fiber probe of the present application can achieve ultrahigh spatial detection resolution, and can be used for detecting nanometer-scale bacteria, viruses, biological macromolecules, etc.
The sample 900 to be detected in this embodiment may be a nano-scale biomacromolecule, virus or pathogenic bacterium, which is widely distributed in human body and nature, and is convenient for detecting diseases and detecting health status of human or other animals; the raman spectral signals received in raman spectrometer 100 may be displayed and further processed by a computer.
The manufacturing method of the monomolecular Raman fiber optical tweezers based on the core-shell micro lens is convenient and quick to operate, detection can be completed without introducing exogenous substances after the preparation is completed, and the monomolecular Raman fiber optical tweezers have high biocompatibility; the core-shell micro-lens 510 optical fiber probe is based on the preparation of the optical fiber probe, and due to the miniaturization of the optical fiber probe, the single-molecule Raman spectrum detection can be carried out in a narrow biological environment; the core-shell micro-lens 510 optical fiber probe has the capability of capturing a single biomolecule, can amplify a Raman scattering signal of the molecule, and can be used for in-situ detection of biomacromolecules or nano-sized bacteria or viruses in a biological environment.
The invention also relates to application of the monomolecular Raman fiber optical tweezers based on the core-shell micro lens, which is applied to the detection of macromolecular organisms.
The optical fiber tweezers and the core-shell micro lens 510 are combined, the optical fiber tweezers and the core-shell micro lens 510 can generate a sub-wavelength optical focal length, the interaction between light and a substance can be enhanced, a generated photopotential trap can more stably capture nano-scale biomolecules, simultaneously, raman scattering signals generated by the molecules are amplified through a echo wall resonance effect generated by the core-shell micro lens 510, and the amplified Raman signals are transmitted to the Raman spectrometer 100 through optical fibers, so that the in-situ detection of the biological single molecules is realized. The invention can be used for capturing and detecting DNA, protein molecules and other biological macromolecules, solves the problem of weak Raman scattering model of the detection target through the echo wall resonance effect, and has good application potential in the field of biomedicine.
The embodiments of the present invention have been described in detail with reference to the drawings, but the present invention is not limited to the embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the gist of the present invention.
Claims (10)
1. A monomolecular Raman fiber optical tweezers based on a core-shell microlens is characterized by comprising:
a Raman spectrometer (100);
a fiber coupler (200), the Raman spectrometer (100) being connected to a first end of the fiber coupler (200);
a fiber laser (300), the fiber laser (300) being connected to a second end of the fiber coupler (200) through an isolator (600);
the third end of the optical fiber coupler (200) is connected with the interface end of the optical fiber probe (400);
the core-shell micro-lens suspension liquid (500) is used for containing a sample (900) to be detected, and the front end of the optical fiber probe (400) extends into the core-shell micro-lens suspension liquid (500).
2. The monomolecular raman fiber optical tweezers based on the core-shell microlens as claimed in claim 1, wherein: the wavelength of the laser emitted by the optical fiber laser (300) is 785nm.
3. The monomolecular raman fiber optical tweezers based on core-shell microlenses according to claim 1, wherein: the core-shell microlens suspension (500) is TiO2/SiO2 core-shell composite particle suspension.
4. The monomolecular raman fiber optical tweezers based on core-shell microlenses according to claim 1, wherein: the diameter of the optical fiber probe (400) is 3-5 mu m, and the cone angle of the optical fiber probe (400) is 60-73 degrees.
5. A manufacturing method of monomolecular Raman fiber optical tweezers based on core-shell microlenses is characterized by comprising the following steps:
acquiring a fiber probe (400), connecting the fiber probe (400) to a third end of a fiber coupler (200), connecting a Raman spectrometer (100) to a first end of the fiber coupler (200), and connecting a fiber laser (300) to a second end of the fiber coupler (200) through an isolator (600);
preparing a core-shell microlens suspension (500);
and dripping the core-shell microlens suspension (500) onto a glass slide, and extending the front end of the optical fiber probe (400) into the core-shell microlens suspension (500).
6. The method for manufacturing monomolecular Raman fiber optical tweezers based on core-shell microlenses according to claim 5, wherein the method comprises the following steps: the manufacturing method of the optical fiber probe (400) comprises the following steps
Cutting the optical fiber into two sections from the middle, stripping off the plastic outer surface skin and the coating layer of the middle section of the optical fiber to obtain a section of bare optical fiber, and sleeving the optical fiber into the metal thin tube;
the bare optical fiber is melted by a carbon dioxide laser, and then the melted portion is attenuated, thereby drawing an optical fiber probe (400).
7. The method for manufacturing monomolecular Raman fiber optical tweezers based on core-shell microlenses according to claim 5, wherein the method comprises the following steps: the specific steps for preparing the core-shell microlens suspension (500) are
Dispersing a titanium precursor in absolute ethyl alcohol to obtain a titanium precursor solution;
adding SiO2 microsphere powder into absolute ethyl alcohol to obtain an ethyl alcohol dispersion liquid of SiO2 microspheres;
stirring the titanium precursor solution, dropwise adding the titanium precursor solution into the ethanol dispersion liquid of the SiO2 microspheres, and then putting the mixture into a constant-temperature reaction container for reaction to obtain core-shell particle ethanol dispersion liquid;
and taking out the core-shell particle ethanol dispersion liquid and diluting the core-shell particle ethanol dispersion liquid by deionized water to obtain core-shell micro-lens suspension liquid (500).
8. The method for manufacturing monomolecular Raman fiber optical tweezers based on core-shell microlenses according to claim 7, wherein the method comprises the following steps: the reaction temperature in the reaction vessel is any value of 28-42 ℃.
9. The method for manufacturing monomolecular Raman fiber optical tweezers based on core-shell microlenses according to claim 7, wherein the method comprises the following steps: the diameter of the SiO2 microsphere powder is 3-5 μm.
10. The method for manufacturing monomolecular Raman fiber optical tweezers based on core-shell microlenses according to claim 7, wherein the method comprises the following steps: the thickness of the core shell in the core shell particle ethanol dispersion liquid is 0.05-0.1 mu m.
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| CN116477563A (en) * | 2023-03-07 | 2023-07-25 | 北京大学 | Packaging method and device for monodisperse microsphere cavity coupling |
| CN116477563B (en) * | 2023-03-07 | 2024-03-12 | 北京大学 | Packaging method and device for monodisperse microsphere cavity coupling |
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