CN110596100A - Bloch wave nano microscopic imaging device based on coaxial double-waveguide optical fiber - Google Patents
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Abstract
The invention provides a Bloch wave nano microscopic imaging device based on a coaxial double-waveguide optical fiber. The method is characterized in that: the coaxial double-waveguide optical fiber laser comprises a coaxial double-waveguide optical fiber 1, a coaxial double-waveguide optical fiber connector 2, a camera 3, a computer 4, a light source 5 and an optical fiber 6. The surface of the multilayer dielectric film substrate 1 can be excited by a large enough wave vector to generate a surface Bloch wave, the surface Bloch wave is sensitive to environmental changes, the surface Bloch wave is excited by laser by utilizing the annular core 1-1 of the coaxial double-waveguide optical fiber 1 to interact with a sample to eliminate the tail caused by the surface plasmon acting on the sample, meanwhile, the middle core of the coaxial double-waveguide optical fiber 1 is used for collecting scattered light, and an image with high signal-to-noise ratio and high resolution ratio is obtained by processing through the computer 4. The invention uses special optical fiber, can effectively reduce cost, optimize structure, and realize portable high signal-to-noise ratio and high resolution micro-nano microscopic imaging.
Description
(I) technical field
The invention relates to a Bloch wave nano microscopic imaging device based on a coaxial double-waveguide optical fiber, belonging to the field of optical fiber microscopic imaging.
(II) background of the invention
Optical microscopy is the most common and effective tool in optical inspection for scientific research. With conventional microscopes, the minimum distance between two objects that can be clearly resolved depends on the limits of the microscope. In order to better visualize the microscopic world, researchers have endeavored to develop various methods of improving resolution. The super-resolution imaging technology has also made a breakthrough, and typically represents Confocal Microscopy (Confocal Microscopy), stimulated emission Depletion Microscopy (STED), Photo-activated localization Microscopy (PALM), and so on. It should be noted that in all super-resolution imaging techniques herein, the optical path used is a far-field leakage radiation imaging system.
Fluorescence detection is an important tool in the field of biological science. Surface-tethered technologies are often used in clinical diagnostics and DNA analysis to capture antibodies, DNA oligomers or target molecules, etc. Fluorescence detection and imaging depend on the location of the excitation light field. For example, evanescent waves generated by Total Internal Reflection (TIR) as proposed by In single-motion imaging with organic detection using object Total internal reproduction contact and Eyen irradiation In Total internal reproduction luminescence using 1-enzyme light can be used for light field surface imaging. The measurement by the TIR method is that the incident light is required to be more than the critical angle, an evanescent field is excited, the longitudinal penetration depth of the evanescent field is about 100nm, and the evanescent field is a local electromagnetic field. This localized electromagnetic field allows selective observation of biomolecules on the surface of the sample, a technique that is critical to the optical field of cell and molecular biology. TIR illumination can be used for selectively imaging the surface of the sample, so that the bulk background signal is reduced to the maximum extent, and the signal-to-noise ratio is improved.
Many methods based on surface signal detection cannot collect fluorescence signals weakly bound to the surface. However, for many types of biological imaging experiments, detection of the sample bulk radiation signal can also provide a useful signal, thus requiring measurement of both the tightly bound molecular fluorescence signal and the bulk target molecular signal. In these cases, it is useful to selectively excite surface or bulk target molecules. The surface bound fluorescence signal measurement allows the bulk signal to be maximally not collected, thus eliminating the step of washing unbound fluorophores. TIR, however, has difficulty in obtaining electromagnetic fields with long evanescent depths and thus in detecting bulk phase signals. Fluorescence microscopy is a typical means of wide field illumination or volumetric imaging away from the surface of a glass substrate. Surface or bulk sample imaging can be achieved using Total Internal Reflection Fluorescence Microscopy (TIRFM) and fluorescence microscopy, respectively, however, simultaneous concurrent bursts of both imaging are difficult to achieve. The simultaneous switching between the two imaging techniques requires precise mechanical alignment, which is difficult to perform in practice.
The surface wave microscope solves the problems, and the surface wave microscope uses surface waves, mainly surface plasmon resonance of metal and air cross section, as an illumination light source, and realizes high-sensitivity imaging of a sample close to the surface of a metal film layer by using the characteristics of strong locality of surface propagation and very sensitivity to disturbance at an interface. Chinese patent CN 103837499A proposes a micro-area spectrum measuring device based on broadband surface plasma waves, which mainly uses a high numerical aperture microscope objective and broadband radial polarized light or radial polarized white light to build up a spectrum measuring device. High spatial resolution can be achieved on the basis of this arrangement. Chinese patent CN 105628655 a proposes an optical microscope based on surface plasmon resonance with high resolution and without fluorescent label, which excites plasmon surface resonance at a plasmon resonance sensing chip, thereby obtaining high spatial resolution. The above-mentioned main microscopic techniques have great limitations in practical applications, and have the following problems:
1. the signal-to-noise ratio is poor. When the traditional surface wave microscopic imaging is carried out, because the surface wave of an excitation field and the surface wave scattered by a sample can interfere with each other, a strong tail can be formed on one side of the sample along the excitation direction, the tail length is equal to the attenuation length of the surface wave along the surface, the tail signal and the sample scattering signal are leaked together and collected by an imaging system, and the imaging signal-to-noise ratio is obviously reduced.
2. The spatial resolution is poor. Also due to smear, when a conventional surface wave imaging system images an actual sample having a boundary, streaky smear is generated at the boundary, which significantly degrades the resolution.
3. The time resolution is poor. In order to improve the resolution, the surface wave imaging system developed in recent years usually needs to acquire images in multiple lengths and multiple angles, and then eliminates the imaging trailing by using an algorithm to improve the resolution. The problems that each microscopic image needs a lot of time and the time resolution is poor, and real-time observation cannot be carried out are brought.
4. The working environment is single and the cost is high. The traditional surface wave imaging system only uses one metal film as a base, and the metal film is used as an imaging substrate, has special requirements on working environment, cannot work in water, is easy to oxidize, cannot be recycled and has higher cost.
Chinese patent CN 109239020a proposes a surface wave imaging system based on rotation illumination, which eliminates the tail caused by the surface wave acting on the sample by the galvanometer scanning system, and improves the signal-to-noise ratio and resolution of the surface wave microscopic imaging, but the used devices are of many kinds and have large volume, resulting in heavy weight and inconvenience.
The single optical fiber imaging technology adopts a single multimode optical fiber for imaging, the optical fiber is an imaging device and an image transmission device, a scene in a section of view field range of the optical fiber can be transmitted to the other end at one time without adding a scanning device and an imaging lens, and the single optical fiber imaging technology belongs to a wide-field optical fiber imaging technology. Through the development of the last 10 years, the single fiber imaging technology has made great progress in the aspects of imaging mechanism, imaging quality, application research and the like, but has many defects in the aspects of imaging speed and resolution.
The invention discloses a Bloch wave nano microscopic imaging device based on a coaxial double-waveguide optical fiber. The defects of low signal-to-noise ratio, poor time and space resolution and high cost of the traditional surface wave imaging microscope are overcome. The coaxial double-waveguide optical fiber is adopted, the surface Bloch wave is used for acquiring scattering signals with high resolution and high signal-to-noise ratio, and an optical fiber imaging mode is adopted for acquiring images; the excitation of the Bloch wave and the acquisition of the image signal are finished by using the same optical fiber, so that a high-quality image can be obtained, and portable microscopic detection imaging can be realized.
Disclosure of the invention
The invention aims to provide a surface enhanced nano microscopic auxiliary device which has simple structure, good stability, low manufacturing cost and easy assembly, can realize the real-time observation of a non-fixed surface sample and obtain a coaxial double waveguide fiber Bloch wave nano microscopic imaging device based on the high signal-to-noise ratio and high resolution imaging of the surface sample.
The purpose of the invention is realized as follows:
the nanometer microscopic imaging device based on coaxial double-waveguide fiber Bloch wave consists of a coaxial double-waveguide fiber 1, a coaxial double-wave optical fiber connector 2, a camera 3, a computer 4, a light source 5 and a fiber 6; the method is characterized in that: a beam of light emitted by a light source 5 passes through a coaxial double-wave optical fiber connector 2 through an optical fiber 6 and is injected into an annular core 1-1 of a coaxial double-wave optical fiber 1, and a micro-machined optical fiber end enables linearly polarized light to form a specific angle with a multi-layer dielectric film substrate on the end face of a cone frustum of the optical fiber, wherein the angle can be obtained under known parameters; laser forms a bundle of light illumination sample illumination samples with specific incidence angle through the annular core 1-1 of the coaxial double waveguide fiber 1, and the light illumination sample illumination samples have enough large wave vectors to effectively excite surface Bloch waves existing in the multilayer dielectric film substrate; when the surface Bloch wave propagates through a sample, scattered signal light and surface tailing are emitted, the laser is incident from the annular core 1-1 of the coaxial double-waveguide fiber 1, and simultaneously surface plasmons are excited by 360 degrees, so that the surface tailing can be effectively eliminated; scattered light is collected by the middle core 1-2 of the coaxial double waveguide fiber 1 connected with the camera 3, forms an interference pattern with the other light of the light source on the camera 3, and the assignment and the phase of the image are extracted from the fiber by the computer 4 by using an off-axis digital holographic algorithm and are restored on the computer, so that high-resolution and contrast surface wave microscopic imaging can be obtained.
The annular core 1-1 of the coaxial double-waveguide optical fiber 1 is an optical fiber which is provided with an annular waveguide which is symmetrical along the axis and a fiber core with a large core diameter in the middle. As shown in FIG. 2, 1-1 is the annular core of the coaxial double-waveguide fiber, and 1-2 is the intermediate core of the coaxial double-waveguide fiber. The intermediate core has a large diameter, scatters signal light, and contains an optical field of image information. Information including intensity distribution, phase distribution, light beam wave front and the like is input into a computer 4 through a camera 3 and is processed and transformed to obtain an image.
The imaging mode of the coaxial double-waveguide fiber Bloch wave nano microscopic imaging device uses the principle of single multimode fiber imaging. The Imaging principle of a Single Multimode Fiber is described in detail in the document "Scanner-Free and Wide-field endoscopic Imaging by Using a Single Multimode Optical Fiber", as shown in fig. 7, a laser emitted by a laser splits light into two parts, the transmitted light is reflected by a mirror and coupled with the Fiber through a mirror BS2, then the other end of the Fiber is a plane to be measured OP to illuminate a target object, then the light collected by the Fiber returns to an IP and then enters a camera to be combined with the light which is split by B1 and reflected into the camera through BS3 to form an interference image on the camera. And extracting the assignment and the phase of the image from the optical fiber by using an off-axis digital holographic algorithm, and restoring the image on a computer.
The imaging mode based on the coaxial double-waveguide fiber Bloch wave nanometer microscopic imaging device is optimized and improved on the basis of the single multimode fiber imaging, the coaxial double-waveguide fiber can be used for replacing a complex light path in a figure (7), a light source 5 divides light into two beams through an optical fiber 6, one beam of light enters an annular core 1-2 of the coaxial double-waveguide fiber 1 through a coaxial double-waveguide fiber connector 2, a surface Bloch wave illumination sample is converged and excited at the other end of the coaxial double-waveguide fiber, scattered light passes through a middle core 1-1 of the coaxial double-waveguide fiber 1, the coaxial double-waveguide fiber connector and the optical fiber 6 and the other beam of light of the light source 5 form an interference image on a camera 6, and assignment and phase of the image are extracted from the fiber by utilizing an off-axis digital holographic algorithm to restore the image on a computer.
The multilayer dielectric film substrate is formed by plating multilayer non-metallic media on the end face of the coaxial double-waveguide optical fiber, is not easy to oxidize and denature, and can be repeatedly cleaned and used.
The multilayer dielectric film substrate supports a Bloch wave mode on the surface through a processed film with the nanometer thickness, and can design a multilayer dielectric nano film supporting different wavelengths and Bloch mode (TE/TM) types as an imaging substrate by changing the refractive index and the thickness of each layer. As shown in fig. 4, the characteristics of the surface bloch wave reflectances of the S wave, the P wave, and S + P as a function of the incident angle θ are obtained by theoretical calculation, thereby obtaining the incident angle at which the resonance effect is optimal. From fig. 4, it can be seen that both S-wave and P-wave can generate resonance effect to excite surface bloch wave. We select the resonance peak obtained for the angle of incidence θ, with the best results.
In the system, a coaxial double-waveguide fiber 1 is used, a relation graph of the reflectivity of the surface Bloch wave along with the change of a resonance angle is obtained according to the parameters of the configured multilayer dielectric film substrate 1 and the known ambient refractive index, the optimal incident angle is obtained, and the coaxial double-waveguide fiber 1 with the conical fiber end with the required angle theta is obtained through the micromachining mode shown in the graph (3).
The coaxial double-waveguide fiber-based Bloch wave nano microscopic imaging device is characterized in that a light source is transmitted by using an annular core 1-1 of a coaxial double-waveguide fiber 1 with an angle theta, and laser can excite a surface Bloch wave 360 degrees around the center as shown in a figure (5), so that a trailing image obtained by a single-direction surface Bloch wave enhancement effect is eliminated, and an image with high signal-to-noise ratio and high resolution is finally obtained.
According to the coaxial double-waveguide fiber-based Bloch wave nanometer microscopic imaging device, scattered light enters a camera 3 from a middle core 1-2 of the coaxial double-waveguide fiber, and is processed by a computer 4 to obtain an image with high resolution and high signal-to-noise ratio.
The coaxial double-waveguide fiber-based Bloch wave nano microscopic imaging device can realize portable microscopic imaging.
(IV) description of the drawings
Fig. 1 is a bloch wave nanometer microscopic imaging device based on a coaxial double-waveguide fiber, which comprises a coaxial double-waveguide fiber 1, a coaxial double-wave optical fiber connector 2, a camera 3, a computer 4 and a light source 5.
FIG. 2 is a cross-sectional view of a coaxial double-waveguide fiber, with 1-1 being the annular core of the coaxial double-waveguide fiber and 1-2 being the intermediate core of the coaxial double-waveguide fiber.
FIG. 3 is a drawing of a polishing process of the annular core 1-1 of the coaxial double waveguide optical fiber 1, starting from (a), with the optical fiber and the polishing disk rotating simultaneously to ensure symmetry of the processed optical fiber; (b) is a diagram of the grinding process; (c) is an effect diagram after finishing grinding; (d) is a defined fiber grind angle.
FIG. 4 is a graph of surface bloch wave energy reflectance versus incident wave angle.
Fig. 5 is a schematic diagram of a 360-degree excitation surface bloch wave.
Fig. 6 is a schematic diagram of a coaxial dual-waveguide optical fiber cone, 8 is a multilayer dielectric film, and 7 is a pellet to be tested.
FIG. 7 is a schematic diagram of a single multimode fiber imaging.
(V) detailed description of the preferred embodiments
The invention is further illustrated with reference to the following figures and specific examples.
Fig. 1 is a schematic structural diagram of a coaxial double-waveguide fiber-based bloch wave nano-microscopic imaging device according to the present invention, and the structure includes a coaxial double-waveguide fiber 1, a coaxial double-waveguide fiber connector 2, a camera 3, a computer 4 and a light source 5. The end face structure of the coaxial double waveguide fiber 1 is shown in fig. 2.
Example 1:
firstly, making a multilayer dielectric film substrate 1, namely plating 10 layers of nano dielectric films with alternating refractive indexes on the surface of a glass plate, wherein the material is Si3N4And SiO2. The thickness of each layer is 250nm, 500nm, 200nm, 300nm, 250nm, 100 nm. From the film parameters, the relationship between the surface bloch wave and the incident light angle as shown in fig. 4 is calculated, showing the relationship of S wave, P wave and S + P, respectively. We select the first formant, corresponding to an angle of 34 degrees.
The coaxial double waveguide fiber 1 was polished in the manner shown in fig. 3 with an angle θ of 34 degrees to obtain a taper end with an angle θ.
In the composition: a beam of light emitted by a light source 5 passes through a coaxial double-wave optical fiber connector 2 through an optical fiber 6 and is injected into an annular core 1-1 of a coaxial double-wave optical fiber 1, and a micro-machined optical fiber end enables linearly polarized light to form a specific angle with a multi-layer dielectric film substrate on the end face of a cone frustum of the optical fiber, wherein the angle can be obtained under known parameters; laser forms a bundle of light illumination sample illumination samples with specific incidence angle through the annular core 1-1 of the coaxial double waveguide fiber 1, and the light illumination sample illumination samples have enough large wave vectors to effectively excite surface Bloch waves existing in the multilayer dielectric film substrate; when the surface Bloch wave propagates through a sample, scattered signal light and surface tailing are emitted, the laser is incident from the annular core 1-1 of the coaxial double-waveguide fiber 1, and simultaneously surface plasmons are excited by 360 degrees, so that the surface tailing can be effectively eliminated; scattered light is collected by the middle core 1-2 of the coaxial double waveguide fiber 1 connected with the camera 3, forms an interference pattern with the other light of the light source on the camera 3, and the assignment and the phase of the image are extracted from the fiber by the computer 4 by using an off-axis digital holographic algorithm and are restored on the computer, so that high-resolution and contrast surface wave microscopic imaging can be obtained.
Claims (3)
1. The nanometer microscopic imaging device based on coaxial double-waveguide fiber Bloch wave is characterized in that: the coaxial double-waveguide optical fiber laser comprises a coaxial double-waveguide optical fiber 1, a coaxial double-waveguide optical fiber connector 2, a camera 3, a computer 4, a light source 5 and an optical fiber 6.
In the composition: a beam of light emitted by a light source 5 passes through a coaxial double-wave optical fiber connector 2 through an optical fiber 6 and is injected into an annular core 1-1 of a coaxial double-wave optical fiber 1, and a micro-machined optical fiber end enables linearly polarized light to form a specific angle with a multi-layer dielectric film substrate on the end face of a cone frustum of the optical fiber, wherein the angle can be obtained under known parameters; laser forms a bundle of light illumination sample illumination samples with specific incidence angle through the annular core 1-1 of the coaxial double waveguide fiber 1, and the light illumination sample illumination samples have enough large wave vectors to effectively excite surface Bloch waves existing in the multilayer dielectric film substrate; when the surface Bloch wave propagates through a sample, scattered signal light and surface tailing are emitted, the laser is incident from the annular core 1-1 of the coaxial double-waveguide fiber 1, and simultaneously surface plasmons are excited by 360 degrees, so that the surface tailing can be effectively eliminated; scattered light is collected by the middle core 1-2 of the coaxial double waveguide fiber 1 connected with the camera 3, forms an interference pattern with the other light of the light source on the camera 3, and the assignment and the phase of the image are extracted from the fiber by the computer 4 by using an off-axis digital holographic algorithm and are restored on the computer, so that high-resolution and contrast surface wave microscopic imaging can be obtained.
2. The Bloch wave-based nano-microscopic imaging apparatus based on the coaxial double-waveguide fiber as claimed in claim 1, wherein the coaxial double-waveguide fiber 1 is an optical fiber having an axially symmetric annular waveguide and a core with a large core diameter in the middle.
3. The coaxial double-waveguide fiber Bloch wave-based nano-microscopic imaging device according to claim 1, wherein incident light is transmitted by using the annular core 1-1 of the coaxial double-waveguide fiber 1, and surface Bloch waves can be excited by 360 degrees simultaneously, so that surface tailing of imaging caused by surface plasmons excited in a single direction is eliminated; backward scattered light is collected by the annular core 1-2 of the coaxial double-waveguide fiber 1, and an image with high signal-to-noise ratio and high resolution is obtained by adopting a fiber imaging technology.
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| CN111552026A (en) * | 2020-04-10 | 2020-08-18 | 桂林电子科技大学 | Optical fiber and system for human body intervention visual photodynamic therapy |
| CN111603133A (en) * | 2020-04-10 | 2020-09-01 | 桂林电子科技大学 | Intravascular insertion type visual flexible optical fiber surgical tool |
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