A nano-imaging system and method of use thereof

Abstract

The application discloses a kind of nano imaging systems and its application method, system includes laser, acoustooptic deflector, beam expander, dichroic mirror, microscope, sample chamber, illumination light source, reflector, condenser, camera and computer, sample substrate of the sample chamber is coated with the sample solution comprising upconversion nanoparticle;Wherein, the light emitted by laser is modulated in turn by acoustooptic deflector, collimated by beam expander, reflected by dichroic mirror, focused by microscope, for capturing and controlling upconversion nanoparticle;The light emitted by illumination light source is projected in turn by sample, dichroic mirror, reflected by reflector, condensed by condenser, and imaged in camera;Computer is used to control laser, acoustooptic deflector and display the image collected by camera.The image resolution of the embodiment of the application can reach nanometer level, and can be widely applied in optical technology field.

Description

Technical Field

[0001] This invention relates to the field of optical technology, and in particular to a nano-imaging system and its application method. Background Technology

[0002] The development of nanopixels can transform various applications in information science and technology, including ultra-high resolution imaging and display, large-capacity data storage, and optical information encryption. In particular, the emergence of the Internet of Things, 5G mobile networks, and virtual / augmented reality has led to an increasing demand for light field, near-eye, and portable / wearable displays with high spatial resolution. Therefore, developments in these fields will greatly benefit from replacing traditional micrometer-scale pixels with nanopixels. Summary of the Invention

[0003] In view of this, the purpose of this invention is to provide a nanoscale imaging system and its application method, with image resolution reaching the nanometer level.

[0004] In a first aspect, embodiments of the present invention provide a nanoimaging system, comprising a laser, an acousto-optic deflector, a beam expander, a dichroic mirror, a microscope, a sample chamber, an illumination source, a reflector, a condenser, a camera, and a computer. The sample substrate in the sample chamber is coated with a sample solution containing upconversion nanoparticles. Light emitted from the laser is sequentially modulated by the acousto-optic deflector, collimated by the beam expander, reflected by the dichroic mirror, and focused by the microscope to capture and control the upconversion nanoparticles. Light emitted from the illumination source is sequentially passed through the sample, projected by the dichroic mirror, reflected by the reflector, and focused by the condenser before being imaged by the camera. The computer is used to control the laser, the acousto-optic deflector, and to display the images acquired by the camera.

[0005] Optionally, the sample solution may also include doped lanthanide ions.

[0006] Optionally, the sample substrate may include any one of a flexible polymer, a flexible biological leaf, an ex vivo skin tissue, and an ex vivo single cell.

[0007] Secondly, embodiments of the present invention provide an application method for a nano-imaging system, including:

[0008] Prepare a sample solution containing upconversion nanoparticles;

[0009] The sample solution is dropped onto the sample substrate, and the sample substrate is placed in the sample chamber;

[0010] By controlling the optical parameters of the laser and the modulation parameters of the acousto-optic deflector with a computer, specific images can be captured and controlled to form upconversion nanoparticles.

[0011] Images of the sample substrate are acquired using a camera and displayed on a computer.

[0012] Optionally, the preparation of the sample solution comprising upconversion nanoparticles specifically includes:

[0013] Upconversion nanoparticles are coated with a silica film of a predetermined thickness;

[0014] Upconversion nanoparticles with a predetermined concentration of silica film were prepared, and a dispersion solution was prepared by shaking to obtain a sample solution.

[0015] Optionally, the optical parameters of the laser and the modulation parameters of the acousto-optic deflector are controlled by a computer to capture and control the upconversion nanoparticles to form specific images, specifically including:

[0016] By controlling the power of the laser and the capture position, deflection angle, and polarization direction of the acousto-optic deflector via computer, different upconversion nanoparticles can be captured and controlled to form specific images at specific locations and times.

[0017] Optionally, the method further includes:

[0018] Dynamic images are formed by controlling the scanning frequency of the acousto-optic deflector with a computer.

[0019] Optionally, the method further includes:

[0020] Determine the correspondence between encrypted information and QR code patterns;

[0021] Based on the encrypted content and its corresponding relationship, the laser and acousto-optic deflector are controlled to move the upconversion nanoparticles at a preset frequency to form a QR code pattern corresponding to the encrypted content.

[0022] Optionally, the method further includes:

[0023] The camera captures QR code patterns at different times according to a preset frequency;

[0024] The encrypted content is obtained based on the QR code pattern and its corresponding relationship.

[0025] Implementing the embodiments of the present invention has the following beneficial effects: The nanoimaging system in this embodiment includes a laser, an acousto-optic deflector, a beam expander, a dichroic mirror, a microscope, a sample chamber, an illumination source, a reflector, a condenser, a camera, and a computer. The sample substrate in the sample chamber is coated with a sample solution containing upconversion nanoparticles. The optical parameters of the laser and the modulation parameters of the acousto-optic deflector are controlled by the computer to capture and control the upconversion nanoparticles to form a specific image. The image of the sample substrate is acquired by the camera, thereby generating an image with nanoscale resolution. Attached Figure Description

[0026] Figure 1This is a structural block diagram of a nanoimaging system provided in an embodiment of the present invention;

[0027] Figure 2 This is a schematic diagram illustrating different patterns formed on different sample substrates according to an embodiment of the present invention;

[0028] Figure 3 This is a flowchart illustrating the steps of an application method for a nano-imaging system provided in an embodiment of the present invention;

[0029] Figure 4 This is a schematic diagram illustrating the encryption and decryption of a nanoimaging system provided in an embodiment of the present invention. Detailed Implementation

[0030] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments. The step numbers in the following embodiments are only for ease of explanation and do not limit the order of the steps. The execution order of each step in the embodiments can be adapted according to the understanding of those skilled in the art.

[0031] In the following description, references are made to “some embodiments,” which describe a subset of all possible embodiments. However, it is understood that “some embodiments” may be the same subset or different subsets of all possible embodiments and may be combined with each other without conflict.

[0032] In the following description, the terms "first, second, third" are used merely to distinguish similar objects and do not represent a specific ordering of objects. It is understood that "first, second, third" may be interchanged in a specific order or sequence where permitted, so that the embodiments of the invention described herein can be implemented in an order other than that illustrated or described herein.

[0033] Unless otherwise defined, all technical and scientific terms used in the embodiments of this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in the embodiments of this invention is for descriptive purposes only and is not intended to limit the invention.

[0034] In a first aspect, embodiments of the present invention provide a nanoimaging system, comprising a laser, an acousto-optic deflector, a beam expander, a dichroic mirror, a microscope, a sample chamber, an illumination source, a reflector, a condenser, a camera, and a computer. The sample substrate in the sample chamber is coated with a sample solution containing upconversion nanoparticles. Light emitted from the laser is sequentially modulated by the acousto-optic deflector, collimated by the beam expander, reflected by the dichroic mirror, and focused by the microscope to capture and control the upconversion nanoparticles. Light emitted from the illumination source is sequentially passed through the sample, projected by the dichroic mirror, reflected by the reflector, and focused by the condenser before being imaged by the camera. The computer is used to control the laser, the acousto-optic deflector, and to display the images acquired by the camera.

[0035] See Figure 1 The nano-imaging system comprises a laser 1-1, an acousto-optic deflector 1-2, a beam expander 1-3, a dichroic mirror 1-4, a microscope 1-5, a sample chamber 1-6, an illumination source 1-7, a reflector 1-8, a condenser 1-9, a camera 1-10, and a computer 1-11. The laser 1-1, acousto-optic deflector 1-2, beam expander 1-3, and dichroic mirror 1-4 are positioned on the same horizontal plane. The microscope 1-5 and sample chamber 1-6 are positioned above the dichroic mirror 1-4. The reflector 1-8 is positioned directly below the dichroic mirror 1-4. The condenser 1-9 is positioned to the left of the reflector 1-8, and the camera 1-10 is positioned to the left of the condenser 1-9.

[0036] The laser is a continuous laser used to capture, manipulate, and stimulate upconversion particles. An acousto-optic deflector controls the spatial position, intensity, and scanning frequency of the captured laser beam. Based on the deflection principle, the acousto-optic deflector can change the laser angle according to the acousto-optic interaction mechanism, thus allowing computer-controlled scanning of the captured laser beam in space. A beam expander collimates the laser beam emitted by the laser (because the emitted laser has a certain divergence angle). A dichroic mirror reflects the collimated laser beam into a microscope. The beam, after passing through a high-magnification microscope, has highly focused characteristics and can be used to capture upconversion nanoparticles in solution. The refractive index of the upconversion nanoparticles is approximately 1.6. The upconversion nanoparticles in the solution are captured by the gradient force of the captured light and are captured to the center of the beam. An illumination source is placed above the inverted microscope. The illumination source passes through the inverted microscope and a dichroic mirror. The dichroic mirror only reflects light of a specific wavelength (such as a 1064nm laser). Therefore, the illumination source passes directly through the dichroic mirror, and then is reflected and focused by a mirror to the CCD camera. The CCD camera records the images and videos presented in the microscope.

[0037] The computer is connected to the acousto-optic deflector, and the entire process is monitored in real time by a CCD camera and displayed synchronously on the computer screen. All monitoring and operation of the experiment is handled by the Tweez program. Because the acousto-optic deflector is directly connected to the computer, the position of the captured light, laser power, scanning frequency, and direction of motion can be directly set in the program. The principle is that the software controls the acousto-optic deflector to a specific angle based on the user-set position of the captured light, focusing the laser onto the set position. When multiple capture points are set, the acousto-optic deflector continuously deflects between corresponding angles according to the order in which they are set. The focused beam can continuously scan between the set capture points, modulating their spatiotemporal distribution (the conversion frequency is adjustable in the software, ranging from 100Hz to 100kHz). Simultaneously, particles are captured and manipulated. Simply moving the sample to the spatial position of the capture trap after capture allows for real-time particle movement and control.

[0038] Optionally, the sample solution may also include doped lanthanide ions.

[0039] The doped lanthanide ions exhibit ionic resonance with the trapped light, enhancing the photodynamic interaction, thus stably trapping the particles at the center of the beam. The upconversion nanoparticles, due to the anti-Stokes phenomenon, absorb the trapped light energy and are excited to emit corresponding fluorescence. (Tm-doped) 3+ Er 3+ Ho 3+ Upconversion nanoparticles of ions emit blue, green, and red fluorescence under excitation light. The doping concentration of lanthanide ions ranges from 15% to 25%. A method utilizing near-infrared light to capture upconversion nanoparticles and simultaneously excite their fluorescence, using the same laser beam for both capture and excitation, reduces the complexity of the instrument system. Adjusting the incident light power and different fluorescent upconversion nanoparticles enables full-color tunable imaging within the visible light range. For example, using a 1064nm laser beam for both excitation and capture of the upconversion nanoparticles reduces the complexity of the experimental system and minimizes photodamage to biological tissues or cells. The emission wavelengths of the upconversion nanoparticles are 650nm, 532 / 650nm, and 450 / 375nm, respectively.

[0040] Optionally, the sample substrate may include any one of a flexible polymer, a flexible biological leaf, an ex vivo skin tissue, and an ex vivo single cell.

[0041] See Figure 2 , Figure 2 a represents bright-field and dark-field fluorescence images of upconversion nanoparticles captured on a flexible PDMS (Polydimethylsiloxane) substrate and collected by a camera in the patterned arrangement. Figure 2b represents a schematic diagram of writing upconversion nanoparticle pattern information onto the surface of biological leaf tissue, pork tissue, and single cell surface, respectively. Figure 2 c is Figure 2 Figure b shows the experimental dark-field fluorescence image; Figure 2 d is a dark-field fluorescence image of the dynamic color display of manipulated upconversion nanoparticles; Figure 2 e is a dark-field fluorescence image of manipulating upconversion nanoparticles to form a "rocket" pattern and move in the y direction; Figure 2 f is a dark-field scattering plot showing the dynamic transformation of the upconversion nanoparticle pattern in the z-direction.

[0042] Implementing the embodiments of the present invention has the following beneficial effects: The nanoimaging system in this embodiment includes a laser, an acousto-optic deflector, a beam expander, a dichroic mirror, a microscope, a sample chamber, an illumination source, a reflector, a condenser, a camera, and a computer. The sample substrate in the sample chamber is coated with a sample solution containing upconversion nanoparticles. The optical parameters of the laser and the modulation parameters of the acousto-optic deflector are controlled by the computer to capture and control the upconversion nanoparticles to form a specific image. The image of the sample substrate is acquired by the camera, thereby generating an image with nanoscale resolution.

[0043] See Figure 3 This invention provides an application method for a nano-imaging system, comprising:

[0044] S100. Prepare a sample solution containing upconversion nanoparticles.

[0045] In this embodiment, the sample solution used is a dispersion solution of silica-encapsulated upconversion nanoparticles with a core-shell structure. The sample solution can also be doped with Tm. 3+ Er 3+ Ho 3+ Ion upconversion nanoparticles (matrix NaYF4:Gd,Yb).

[0046] Optionally, the preparation of the sample solution comprising upconversion nanoparticles specifically includes:

[0047] S110. The upconversion nanoparticles are coated with a silica film of a predetermined thickness.

[0048] S120. Prepare upconversion nanoparticles with a preset concentration of silica film coating, and prepare a dispersion solution by shaking to obtain a sample solution.

[0049] First, upconversion nanoparticles were coated with a silica film of a predetermined thickness. The upconversion nanoparticles consisted of a core-shell structure encapsulated in dense silica, with a size of approximately 35 nm. The silica film was 10 nm thick, serving to impart good hydrophilicity to the upconversion nanoparticles, allowing them to be directly dispersed in aqueous solutions and improving their biocompatibility. Next, an upconversion nanoparticle solution with a concentration of 0.04 mg / mL was prepared. At room temperature, the prepared solution was placed in an ultrasonic shaker and agitated at a frequency of 20–80 kHz for 2–10 minutes to prepare an upconversion nanoparticle dispersion. Then, a suitable amount of the prepared upconversion nanoparticle dispersion was taken out, and its morphology was observed using a transmission electron microscope. The upconversion nanoparticles exhibited a uniform spherical structure, ensuring uniform pixel size in the image, with a size of approximately 35 nm.

[0050] S200. Drop the sample solution onto the sample substrate and place the sample substrate in the sample chamber.

[0051] Specifically, the sample solution can be coated onto the sample substrate using a pipette. In one specific embodiment, a glass slide or PDMS is fabricated into a sample substrate with a length of 0.5 cm, a width of 0.2 cm, and a height of 0.1 cm. 10 μL of the prepared upconversion nanoparticle dispersion solution is taken out using a pipette, and then the upconversion nanoparticle suspension is injected into the sample substrate. Finally, the sample substrate is placed in the corresponding position in the sample cell.

[0052] S300 uses computer-controlled optical parameters of the laser and modulation parameters of the acousto-optic deflector to capture and control upconversion nanoparticles to form specific images.

[0053] The optical parameters of a laser include, but are not limited to, laser power or laser wavelength, and the modulation parameters of an acousto-optic deflector include, but are not limited to, capture position, deflection angle, and polarization direction.

[0054] Optionally, the optical parameters of the laser and the modulation parameters of the acousto-optic deflector are controlled by a computer to capture and control the upconversion nanoparticles to form specific images, specifically including:

[0055] S310. The power of the laser is controlled by a computer, and the capture position, deflection angle and polarization direction of the acousto-optic deflector are controlled by a computer to capture and control different upconversion nanoparticles to form specific images at specific positions and at specific times.

[0056] Specifically, the spatial location of the capture point and the laser power (wavelength: 1064nm, power: 200mW) are set. Due to the control of the acousto-optic deflector, a single beam scans rapidly along a certain path (the capture point movement path code is written using Tweez 250si-TCP Server). The Gaussian beam applies an optical gradient to capture and arrange the upconversion nanoparticles in the solution into a static pattern.

[0057] Optionally, the method further includes:

[0058] S320: The scanning frequency of the acousto-optic deflector is controlled by a computer to form a dynamic image.

[0059] Upconversion nanoparticles are captured in solution using optical tweezers to form programmable patterns. An acousto-optic deflection system controls the spatial position of the capture point to change over time. Due to the action of the optical gradient force, the programmable pattern of the upconversion nanoparticles moves with the capture point. The captured light excites the fluorescence of the upconversion nanoparticles, and the fluorescence image is dynamically displayed using a dark-field microscope.

[0060] The acousto-optic deflector can rapidly change the customized array of optical potential traps, and the color image composed of upconverted nanopixels captured by optical force changes accordingly, forming video with a maximum frame rate of 200fps.

[0061] S400: Acquires images of the sample substrate using a camera and displays the images on a computer.

[0062] Due to the van der Waals forces between the upconversion particles and the substrate, the static pattern is fixed on the substrate. The camera collects the captured light from the upconversion nanoparticles, which excites the corresponding fluorescence, and the fluorescence pattern is observed on the computer screen.

[0063] Optionally, the method further includes:

[0064] S510. Determine the correspondence between encrypted information and QR code patterns;

[0065] S520: Based on the encrypted content and corresponding relationship, control the laser and acousto-optic deflector to move the upconversion nanoparticles at a preset frequency to form a QR code pattern corresponding to the encrypted content.

[0066] The relationship between encrypted information and QR code patterns is one-to-one; one encrypted message corresponds to one QR code pattern. In a specific embodiment, the Tweez 250si-TCP Server code is first used to write four different QR code patterns for the capture point path. Each QR code pattern contains different numbers, representing different phone numbers, thus completing the encoding of the capture point information.

[0067] The prepared upconversion nanoparticle dispersion solution was taken out and dropped onto a glass slide. Due to the applied light gradient force (capture laser power of 500mW), the upconversion nanoparticles in the solution were captured to form a QR code pattern. (See reference...) Figure 4 In step a, I-IV change the path of the QR code capture point every two seconds, and the upconversion nanoparticle QR code fluorescence image changes accordingly, thus completing the dynamic information encryption in time.

[0068] By utilizing the dynamic change characteristics of upconversion nanopixels, fluorescence information encoding and dynamic encryption in both time and space were achieved.

[0069] Optionally, the method further includes:

[0070] S610: The camera acquires QR code patterns at different times according to a preset frequency;

[0071] S620. Obtain the encrypted content based on the QR code pattern and its corresponding relationship.

[0072] The preset frequency of the camera scanning image is the same as the preset frequency of the acousto-optic deflector encoding, and the encrypted content represents the content information corresponding to the QR code pattern. In a specific embodiment, see [reference needed]. Figure 4 b. First, use code to write the capture point movement path as a QR code (the information is "Pixel"). Then, take out the prepared upconversion nanoparticle dispersion solution and drop it onto a glass slide. Use a computer to set two capture points in the solution. The first capture point is written with a movement path as a QR code (two pixel blocks are taken out from the lower left corner). The other capture point is the two pixel blocks taken out from the lower left corner of the QR code. Its spatial position is (0,0). The movement path is the pixel block in the lower left corner of the QR code. The capture point moves towards the first capture point at a speed of 1 micrometer / second until it reaches (20,0) and forms a complete QR code.

[0073] By utilizing the dynamic change characteristics of upconversion nanopixels, dynamic decryption of fluorescence information in both time and space was achieved.

[0074] It is evident that the content of the above system embodiments is applicable to this method embodiment. The specific functions implemented in this method embodiment are the same as those in the above system embodiments, and the beneficial effects achieved are also the same as those achieved in the above method embodiments.

Claims

1. A nanoimaging system, characterized in that, The system includes a laser, an acousto-optic deflector, a beam expander, a dichroic mirror, a microscope, a sample chamber, an illumination source, a reflector, a condenser, a camera, and a computer. The sample substrate in the sample chamber is coated with a sample solution containing upconversion nanoparticles. Light emitted from the laser is sequentially modulated by the acousto-optic deflector, collimated by the beam expander, reflected by the dichroic mirror, and focused by the microscope to capture and control the upconversion nanoparticles. Light emitted from the illumination source is sequentially projected through the sample, the microscope, the dichroic mirror, reflected by the reflector, and focused by the condenser before being imaged by the camera. The computer controls the laser and the acousto-optic deflector and displays the images acquired by the camera. The sample solution also includes doped lanthanide ions, which generate ion resonance with the captured light, causing the upconversion nanoparticles to be stably captured at the center of the beam. The upconversion nanoparticles absorb the captured light energy through the anti-Stokes phenomenon and are excited to emit corresponding fluorescence.

2. The nanoimaging system according to claim 1, characterized in that, The sample substrate includes any one of flexible polymer, biological flexible leaf, ex vivo skin tissue, and ex vivo single cell.

3. A method for applying a nanoimaging system, wherein the system is used in accordance with the nanoimaging system as described in claim 1, characterized in that, include: Prepare a sample solution containing upconversion nanoparticles; The sample solution is dropped onto the sample substrate, and the sample substrate is placed in the sample chamber; By controlling the optical parameters of the laser and the modulation parameters of the acousto-optic deflector with a computer, specific images can be captured and controlled to form upconversion nanoparticles. Images of the sample substrate are acquired using a camera and displayed on a computer. The doped lanthanide ions resonate with the captured light, causing the upconversion nanoparticles to be stably captured at the center of the beam. The upconversion nanoparticles absorb the captured light energy through the anti-Stokes phenomenon and are excited to emit corresponding fluorescence.

4. The application method according to claim 3, characterized in that, The preparation of the sample solution including upconversion nanoparticles specifically includes: Upconversion nanoparticles are coated with a silica film of a predetermined thickness; Upconversion nanoparticles with a predetermined concentration of silica film were prepared, and a dispersion solution was prepared by shaking to obtain a sample solution.

5. The application method according to claim 3, characterized in that, By controlling the optical parameters of the laser and the modulation parameters of the acousto-optic deflector through computer control, specific images can be captured and controlled to form upconversion nanoparticles. Specifically, this includes: By controlling the power of the laser and the capture position, deflection angle, and polarization direction of the acousto-optic deflector via computer, different upconversion nanoparticles can be captured and controlled to form specific images at specific locations and times.

6. The application method according to claim 5, characterized in that, The method further includes: Dynamic images are formed by controlling the scanning frequency of the acousto-optic deflector with a computer.

7. The application method according to claim 3, characterized in that, The method further includes: Determine the correspondence between encrypted information and QR code patterns; Based on the encrypted content and its corresponding relationship, the laser and acousto-optic deflector are controlled to move the upconversion nanoparticles at a preset frequency to form a QR code pattern corresponding to the encrypted content.

8. The application method according to claim 7, characterized in that, The method further includes: The camera captures QR code patterns at different times according to a preset frequency; The encrypted content is obtained based on the QR code pattern and its corresponding relationship.

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