Use of micro-lens chip in particle tracking

Abstract

The use of a micro-lens chip (10) in particle tracking. A particle tracking apparatus (1) comprising a micro-lens chip (10). A particle tracking method for performing particle tracking by means of a micro-lens chip (10). The use of a micro-lens chip (10) in particle tracking performed by a particle tracking apparatus (1) by means of a particle tracking method.

Description

Use of Microlens Chips in Particle Tracking Technical Field

[0001] The present application relates to the field of optical imaging, and more specifically to the use of microlens chips in particle tracking. Background Art

[0002] Particle tracking technology detects the optical signals (transmitted light, scattered light, quantum dot luminescence or fluorescence) of particles (such as cells, nanoparticles, liposomes, exosomes, bacteria, viruses, molecules, fluorescent molecules, quantum dots, organelles, and nucleic acids) to image and locate the particles and record their trajectories, thereby obtaining dynamic information about the particles. This dynamic information not only reflects the diffusion constant and hydrodynamic radius of the observed or tracked particles, but is also crucial for gaining a deeper understanding of the complex mechanisms behind various physical, chemical, and biological phenomena occurring at the nanoscale.

[0003] Existing particle tracking technologies rely heavily on optical detection of particles. In order to detect the smallest possible optical signal of particles, these particle tracking technologies place high demands on the optical system. Particle tracking technologies based on optical microscopy (such as dark field microscopy, phase contrast microscopy, scattering interferometry microscopy, and surface plasmon resonance imaging microscopy) require dedicated microscope configurations, high numerical aperture lenses, and strong illumination power (typically 1-10 kW / cm 2 ) as well as highly stable laser sources and highly sensitive optical detectors. This makes the microscope system optical path complex and expensive, and in some cases requires sophisticated image processing using software. Surface plasmon-enhanced particle tracking technology requires specific nanophotonic structures and complex near-field configurations, similarly burdened by complex and expensive optical systems.

[0004] All of these limitations make nanoparticle tracking instruments expensive, bulky, and complex to use, making access and widespread adoption of these technologies challenging. Furthermore, these shortcomings hinder their application in other areas, such as sensing and point-of-care diagnostics.

[0005] Summary of the Invention

[0006] In one aspect, the present application provides a particle tracking device comprising a microlens chip.

[0007] In one aspect, the present application provides a particle tracking method, comprising performing particle tracking through a microlens chip.

[0008] In one aspect, the present application provides a particle tracking method, comprising performing particle tracking using the particle tracking device of the present application.

[0009] In one aspect, the present application provides use of a microlens chip in particle tracking.

[0010] In one aspect, the present application provides use of a microlens chip in particle tracking by the particle tracking device of the present application.

[0011] In one aspect, the present application provides use of a microlens chip in particle tracking using the particle tracking method of the present application.

[0012] In one aspect, the present application provides use of a microlens chip in performing particle tracking using the particle tracking device of the present application and the particle tracking method of the present application.

[0013] In one aspect, the present application provides a method for preparing a particle tracking device, comprising preparing a microlens chip.

[0014] In the microlens chip of the present application, the diameter of the microlens is 30-400 microns, more particularly 35-350 microns, more particularly 40-300 microns, more particularly 45-250 microns, more particularly 50-200 microns, more particularly 55-150 microns, more particularly 60-120 microns.

[0015] In some embodiments, the microlenses in the microlens chip of the present application can achieve higher positioning accuracy, thereby enabling more effective monitoring of biochemical processes with smaller spatial variations. In some embodiments, the microlenses of the present application can achieve particle positioning accuracy of less than or equal to 4.0 nanometers, particularly less than or equal to 3.7 nanometers, particularly less than or equal to 3.5 nanometers, and particularly less than or equal to 3.0 nanometers, at a temporal resolution of 1 millisecond.

[0016] In some embodiments, the microlenses in the microlens chip of the present application can achieve higher temporal resolution, enabling more effective monitoring of faster biochemical processes or tracking of faster-moving particles. In some embodiments, the temporal resolution of the microlenses of the present application, when particle positioning accuracy is required to be less than 4 nanometers, is less than or equal to 1.00 milliseconds, particularly less than or equal to 0.90 seconds, particularly less than or equal to 0.80 milliseconds, and particularly less than or equal to 0.50 milliseconds.

[0017] In some embodiments, the microlenses in the microlens chip of the present application have a relatively high adjustable focal length range, which is beneficial for tracking particles within a larger range in the solution.

[0018] In some embodiments, the microlenses in the microlens chip of the present application have a stronger ability to converge light and can achieve significant optical signal enhancement, greatly improving the optical signal of the particles and the optical signal received by the objective lens, thereby reducing the requirements for the optical system; thereby reducing the complexity and cost of the optical system, and overcoming the shortcomings of the particle tracking technology optical system being complex and expensive.

[0019] In some embodiments, the particle tracking device of the present application has a simple structure, low cost, and good stability; the particle tracking method of the present application has a simple structure, low cost, and good stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] FIG1 is a schematic structural diagram of a particle tracking device in some embodiments of the present application.

[0021] Figures 2A and 2B are schematic diagrams of the structure of a microlens chip in some embodiments of the present application. Figure 2C is a schematic diagram of the steps of preparing a microlens chip in some embodiments of the present application.

[0022] FIG3 is a schematic diagram of observing particles through a microlens in some embodiments of the present application.

[0023] Figures 4A, 4B, and 4C show the results of imaging particles in solution using 3-μm, 22-μm, and 73-μm microlenses at different focal lengths, respectively. The value below each image indicates the focal length. Figure 4D shows the adjustable focal length ranges for the 3-μm, 22-μm, and 73-μm microlenses.

[0024] Figure 5A is a simulation diagram of the light-gathering capability of microlenses of different diameters. Figure 5B is a diagram of the light-gathering intensity of microlenses of different diameters.

[0025] Figure 6A shows the results of particle tracking in solution using microlenses of different diameters. Figure 6B shows the optical signal intensity of particles tracked in solution using microlenses of different diameters.

[0026] Figure 7 shows the positioning accuracy of particles under microlenses of different diameters at a time resolution of 1 millisecond.

[0027] Figure 8 shows the time resolution when the particle positioning accuracy is required to be less than 4 nanometers under microlenses of different diameters.

[0028] Figure 9 shows the positioning accuracy of particles under microlenses of different diameters at a time resolution of 1 millisecond.

[0029] FIG10 shows the time resolution of particles under microlenses of different diameters at a time resolution of 1 millisecond.

[0030] The reference numerals in the figures are as follows: 1: particle tracking device; 10: microlens chip; 100: substrate; 110: microlens; 120: coating layer; 20: liquid channel; 200: liquid reservoir; 201, 201a, 201b: particles; 202: detection field of view; 210: liquid inlet and outlet; 30: optical device; 310: objective lens; 320: moving device; 330: optical path; 340: light source; 350: optical sensor; 40: data processing device. DETAILED DESCRIPTION

[0031] In order to further illustrate the technical means and effects adopted by this application to achieve the intended purpose, the specific implementation methods, structures, features and effects of this application are described in detail below in combination with the accompanying drawings and preferred embodiments.

[0032] As used herein, the term "diameter" is used using its conventional definition. For example, when the term "diameter" is used to refer to the diameter of a sphere (e.g., a spherical microlens), it uses the conventional definition of spherical diameter; when the term "diameter" is used to refer to a cylinder (e.g., a cylindrical lens), it refers to the circular diameter of any base of the cylinder, using the conventional definition of circular diameter. The definition of diameter herein is not intended to define a length independent of diameter, such as the height of a cylinder.

[0033] FIG1 is a schematic diagram of a particle tracking device 1 in some embodiments of the present application. The particle tracking device 1 may include a microlens chip 10 .

[0034] Figures 2A and 2B are schematic diagrams of the structure of a microlens chip 10 in some embodiments of the present application. Figure 2A may be a front view, while Figure 2B may be a top view. The microlens chip 10 may include a plurality of microlenses 110, and in particular, may include a substrate 100 and a plurality of microlenses 110 disposed on the substrate 100. In some embodiments, the microlens chip 10 may further include a coating layer 120 disposed on the substrate 100, wherein the coating layer 120 surrounds the microlenses 110.

[0035] Figure 2C is a schematic diagram illustrating the steps of preparing a microlens chip 10 in some embodiments of the present application. In some embodiments, preparing the microlens chip 10 may include: a) providing a substrate 100; and b) disposing a plurality of microlenses 110 on the substrate 100. In some embodiments, preparing the microlens chip 10 may further include c) disposing a coating layer 120 on the substrate 100, wherein the coating layer 120 surrounds the microlenses 110.

[0036] In some embodiments, the substrate 100 may be a transparent substrate, in particular a transparent glass substrate.

[0037] In some embodiments, the microlens 110 may include or consist of a dielectric material, particularly barium titanate. In some embodiments, the microlens 110 is transparent. In some embodiments, the microlens 110 may be a spherical microlens or a cylindrical lens. In some embodiments, the microlens 110 may be an aspherical microlens, particularly an ellipsoidal microlens, a parabolic microlens, or a hyperbolic microlens.

[0038] In some embodiments, the diameter of the microlens 110 is 30-400 microns, more particularly 35-350 microns, more particularly 40-300 microns, more particularly 45-250 microns, more particularly 50-200 microns, more particularly 55-150 microns, more particularly 60-120 microns.

[0039] In some embodiments, the refractive index of the microlens 110 is 1.65-2.20, more particularly 1.80-2.10, more particularly 1.90-2.00, more particularly 1.91-1.93, and more particularly 1.92.

[0040] In some embodiments, in the microlens chip 10, the number of the microlenses 110 is greater than or equal to 1, in particular greater than or equal to 10, in particular greater than or equal to 100, in particular greater than or equal to 10. 2 to 10, more particularly 10 3 to 10.

[0041] In some embodiments, the microlenses 110 are formed on a plane above the substrate 100. In some embodiments, the microlenses 110 are arranged in a hexagonal packing on the plane, and more particularly, in a hexagonal packing on the plane above the substrate 100. In some embodiments, the microlenses 110 contact the substrate 100.

[0042] In some embodiments, the field of view diameter of the microlens 110 is 5 microns to 100 microns. In some embodiments, the pincushion distortion of the field of view of the microlens 110 is within 2% in the central region (30% of the field of view). In some embodiments, the depth of focus of the microlens 110 is 200 nanometers to 600 nanometers.

[0043] In some embodiments, the microlens 110 has a particle positioning accuracy of less than or equal to 4.0 nm, particularly less than or equal to 3.7 nm, particularly less than or equal to 3.5 nm, particularly less than or equal to 3.2 nm, particularly less than or equal to 3.0 nm, at a time resolution of 1 millisecond. In some embodiments, the microlens 110 has a particle positioning accuracy of 2.0-4.0 nm, particularly 2.0-3.7 nm, particularly 2.0-3.5 nm, particularly 2.0-3.2 nm, particularly 2.0-3.0 nm, at a time resolution of 1 millisecond.

[0044] In some embodiments, the time resolution of the microlens 110 when the particle positioning accuracy is required to be less than 4 nanometers is less than or equal to 1.00 milliseconds, particularly less than or equal to 0.90 milliseconds, particularly less than or equal to 0.80 milliseconds, particularly less than or equal to 0.70 milliseconds, particularly less than or equal to 0.60 milliseconds, particularly less than or equal to 0.50 milliseconds. In some embodiments, the time resolution of the microlens 110 when the particle positioning accuracy is required to be less than 4 nanometers is 0.35-1.00 milliseconds, particularly 0.35-0.90 milliseconds, particularly 0.35-0.80 milliseconds, particularly 0.35-0.70 milliseconds, particularly 0.35-0.60 milliseconds, particularly 0.35-0.50 milliseconds.

[0045] In some embodiments, the adjustable focal length range of the microlens 110 is greater than or equal to 1 micron, particularly greater than or equal to 2 microns, particularly greater than or equal to 3 microns, particularly greater than or equal to 4 microns, particularly greater than or equal to 4.5 microns, particularly greater than or equal to 5 microns.

[0046] In some embodiments, the diameter of the microlens 110 and the thickness of the coating layer 120 differ within ±10%, particularly within ±5%, particularly within ±1%, particularly the diameter of the microlens 110 and the thickness of the coating layer 120 are the same.

[0047] In some embodiments, the coating layer 120 has a thickness of 30-400 microns, more particularly 35-350 microns, more particularly 40-300 microns, more particularly 45-250 microns, more particularly 50-200 microns, more particularly 55-150 microns, more particularly 60-120 microns.

[0048] In some embodiments, the coating layer 120 may include a polymer material, particularly a polymer resin material, more particularly polydimethylsiloxane or epoxy resin, more particularly polydimethylsiloxane, or may be composed thereof. In some embodiments, the coating layer 120 is transparent.

[0049] In some embodiments, the microlens 110 and the substrate 100 are optically connected. In particular, two opposite surfaces of the microlens chip 10 can be optically connected to the microlens 110 through the substrate 100 .

[0050] In some embodiments, the method for preparing the particle tracking device 1 includes preparing a microlens chip 10 .

[0051] In some embodiments, preparing the microlens chip 10 includes providing a substrate 100 , such as cleaning and drying the substrate 100 .

[0052] In some embodiments, preparing the microlens chip 10 includes forming the microlenses 110, in particular forming the microlenses 110 on the substrate 100, and in particular including providing, in particular dropping, a suspension including the microlenses 110 and a solvent onto the substrate 100, and removing the solvent, in particular removing the solvent by drying.

[0053] In some embodiments, preparing the microlens chip 10 includes forming a coating layer 120 on a substrate 100, including spin-coating a material for the coating layer 120 onto the substrate 100. The thickness of the coating layer 120 can be controlled by adjusting the coating speed and time, and the ratio of the material for the coating layer 120. In some embodiments, forming the coating layer 120 on the substrate 100 also includes curing the material for the coating layer 120. In some embodiments, the microlenses 110 are first formed on the substrate 100, and then the coating layer 120 is formed on the substrate 100, particularly on the substrate 100 and the microlenses 110, particularly by spin coating. In some embodiments, the coating layer 120 is first formed on the substrate 100, particularly by spin coating, and then the microlenses 110 are formed on the substrate 100, particularly on the substrate 100 and the coating layer 120.

[0054] In some embodiments, the particle tracking device 1 may further include a liquid channel 20. The liquid channel 20 may include a liquid reservoir 200 adjacent to the microlens chip 10, particularly adjacent to the microlens 110. The liquid channel 20 may further include at least one, and particularly at least two, liquid inlets and outlets 210 adjacent to the liquid reservoir 200. In some embodiments, each liquid inlet and outlet 210 communicates with the liquid reservoir 200, allowing liquid to enter the liquid reservoir 200 from one of the liquid inlets and outlets 210 and exit the liquid reservoir 200 from another of the liquid inlets and outlets 210. This allows the particle tracking device 1 of the present application to track particles in liquid continuously flowing in the liquid reservoir 200. In some embodiments, the liquid channel 20 is optically connected to the microlens 110. In some embodiments, the liquid channel 20 may be bonded to the microlens chip 10, particularly to the surface of the microlens 110 and the coating layer 120 of the microlens chip 10. In some embodiments, the method for preparing the particle tracking device 1 includes disposing a liquid channel 20 adjacent to the microlens chip 10 , and in particular, bonding the liquid channel 20 to the microlens chip 10 .

[0055] In some embodiments, the particle tracking device 1 may further include an optical device 30. In some embodiments, the optical device 30 may include an objective lens 310. In some embodiments, the optical device 30 may further include an optical path 330, a light source 340, and an optical detector 350, wherein the optical path 330 optically connects the objective lens 310, the light source 340, and the optical detector 350. In some embodiments, the optical device of the present application is based on a reflective illumination structure. In some embodiments, the optical detector 350 may be electrically connected to a data processing device 40 for processing data received by the optical detector 350.

[0056] In some embodiments, light can originate from the light source 340, pass through the optical path 330, and pass through the objective lens 310 to reach the microlens chip 10; then pass through the microlens chip 10, particularly the substrate 100 and the microlens 110, to reach the liquid in the liquid channel 20. In some embodiments, light can originate from the liquid in the liquid channel 20, particularly the particle 201, pass through the microlens chip 10, particularly the microlens 110 and the substrate 100, to reach the objective lens 310, and then pass through the objective lens 310 and pass through the optical path 330 to reach the optical detector 350. In some embodiments, the optical detector 350 is configured to receive light originating from the particle 201.

[0057] The light emitted from the liquid in the liquid channel 20 can be light of the same wavelength or a different wavelength than the light emitted by the light source 340, in particular, light of the same wavelength or a higher wavelength. The light emitted from the liquid in the liquid channel 20 can be scattered light (such as Rayleigh scattered light or Mie scattered light), or it can be fluorescent light or quantum dot luminescence.

[0058] In some embodiments, the light source 340 may be an LED light source, a halogen light source, a laser light source, or a xenon light source. In some embodiments, the optical path 330 may include a filter. In some embodiments, the optical device may further include a moving device 320 connected to the objective lens 310 or the microlens chip 10 to move the objective lens 310 or the microlens chip 10 to adjust the distance between the objective lens 310 and the microlens chip 10.

[0059] FIG3 is a schematic diagram of a microlens 110 observing particles 201 in some embodiments of the present application. In some embodiments, light emanating from particles 201 in the liquid reservoir 200, for example, light emanating from particles 201 within the detection field 202 corresponding to each microlens 110, can be collected by the microlenses 110, captured by the objective lens 310, and imaged onto the optical sensor 350 through the optical path 330, thereby generating an image of the particles 201 in the far field. Each microlens 110 can generate an image of the particles 201. In some embodiments, the focal plane of the microlens 110 can be changed by adjusting the distance between the objective lens 310 and the microlens chip 10. For example, the focal plane can be shifted to the surface of the microlens 110 to observe particles 201a located on the surface of the microlens 110, or shifted to a certain distance from the microlens (e.g., above the microlens) to observe particles 201b located a certain distance from the microlens (e.g., above the microlens).

[0060] In some embodiments, after the optical signal of particle 201 is received by optical sensor 350, an image can be generated. Data processing of this image (e.g., using, but not limited to, a single particle tracking algorithm) can be performed to obtain the position coordinates of particle 201, and further, a trajectory of its position coordinates over time can be obtained, thereby enabling particle tracking and obtaining other particle-related data. In some embodiments, data processing device 40 can obtain information such as the coordinate position of particle 201 from the image generated by optical sensor 350.

[0061] In some embodiments, particles 201 are, but are not limited to, cells, nanoparticles, liposomes, exosomes, bacteria, viruses, molecules, quantum dots, organelles, and / or nucleic acids. In some embodiments, particles 201 absorb light and emit scattered light, quantum dot luminescence, and / or fluorescence. In some embodiments, particles 201 have an average particle size of 1-1000 nm, particularly 10-500 nm, particularly 20-200 nm, particularly 50-150 nm, particularly 75-125 nm, particularly 90-100 nm, and particularly 100 nm. In some embodiments, particles 201 are dynamic particles. In some embodiments, particles 201 are static particles. In some embodiments, particles 201 are located in the reservoir 200 and contact the microlens 110. In some embodiments, particles 201 are located in the reservoir 200 but do not contact the microlens 110. In some embodiments, the particles 201 are located in the reservoir 200 and are greater than or equal to 0 μm, particularly greater than or equal to 1 μm, particularly greater than or equal to 2 μm, particularly greater than or equal to 3 μm, particularly greater than or equal to 4 μm, particularly greater than or equal to 4.5 μm, particularly greater than or equal to 5 μm, or particularly 0-10 μm, particularly 0-5 μm, particularly 0-3 μm, particularly 0-1 μm away from the microlens 110 .

[0062] Example 1 Relationship between the diameter of a microlens and the adjustable focal length range of the microlens

[0063] Focal length refers to the distance between the center of the lens and the object being imaged, which allows the lens to produce a clear image. The adjustable focal range refers to the range of the lens's focal length relative to the object that can be imaged clearly by adjusting the lens (for example, by adjusting the distance between the lens and the objective lens).

[0064] 3 micron, 22 micron and 73 micron microlenses (all with a refractive index of 1.92) were selected to image the particles in the solution to obtain the adjustable focal length range of each lens. The results are shown in Figures 4A-4D. Figures 4A, 4B, and 4C are the results of imaging particles in the solution with 3 micron, 22 micron and 73 micron microlenses with different focal length ranges, respectively. The value below each picture is the focal length. Figure 4D shows the adjustable focal length range of the 3 micron, 22 micron and 73 micron microlenses. It can be seen that the adjustable focal length range of the 3 micron microlens is approximately 0.15 micron; the adjustable focal length range of the 22 micron microlens is approximately 0.8 micron; the adjustable focal length range of the 73.5 micron microlens is greater than 4.5 microns. As the diameter of the microlens increases, its adjustable focal length range also increases, which is beneficial for tracking particles in a larger range (for example, particles 201, including particles 201 contacting the microlens 110 and particles 201 away from the microlens 110) in the solution (for example, in the liquid in the reservoir 200), and is more conducive to the tracking effect.

[0065] It can be seen that in some embodiments of the present application, microlenses in a specific diameter range have a relatively high adjustable focal length range, which is beneficial for tracking particles within a larger range in the solution.

[0066] Example 2 Relationship between the diameter of a microlens and the adjustable focal length range of the microlens

[0067] Finite element analysis simulation was performed using COMOL software to simulate the light converging ability of microlenses with different diameters.

[0068] Microlenses with diameters of 10 microns, 20 microns, 30 microns, 40 microns, 50 microns, 60 microns and 70 microns (all with a refractive index of 1.92) were selected to simulate their ability to converge light. The results are shown in Figures 5A-5B. Figure 5A is a simulation diagram of the light-gathering ability of microlenses with different diameters. Below each simulation diagram in Figure 5A is the diameter of the microlens. The colors in Figure 5A indicate the intensity of light. The strongest light intensity in each diagram in Figure 5A was selected as the focusing intensity for comparison, as shown in Figure 5B. Figure 5B shows the focusing intensity of microlenses with different diameters. It can be seen that the ability to converge light (focusing intensity) increases with the increase of the diameter of the microlens.

[0069] It can be seen that in some embodiments of the present application, microlenses in a specific diameter range have a stronger ability to converge light, which can improve the optical signal received by the objective lens and reduce the requirements for the optical system; thereby reducing the complexity and cost of the optical system and overcoming the shortcomings of the particle tracking technology's complex and expensive optical system.

[0070] Example 3 Relationship between the diameter of the microlens and the intensity of the optical signal detected by the particle

[0071] Microlenses of 22, 34, 52, 73, and 103 microns (all with a refractive index of 1.92) were selected to track particles in solution. The results are shown in Figure 6A. Figure 6A shows the results of particle tracking in solution using microlenses of different diameters. The intensity of the optical signal was measured, as shown in Figure 6B. Figure 6B shows the optical signal intensity of particles in solution tracked using microlenses of different diameters. It can be seen that the larger the diameter of the microlens, the greater the optical signal intensity of the particle.

[0072] It can be seen that in some embodiments of the present application, microlenses in a specific diameter range can achieve significant optical signal enhancement, greatly improve the optical signal of the particles, and reduce the requirements for the optical system; thereby reducing the complexity and cost of the optical system, and overcoming the shortcomings of the particle tracking technology's complex and expensive optical system.

[0073] In addition, the greater the optical signal intensity of the particles, the higher the positioning accuracy and tracking time resolution of the particle tracking.

[0074] Example 4 Relationship between the diameter of the microlens and the detection particle tracking positioning accuracy and tracking time resolution

[0075] Positioning accuracy is a measure of the accuracy of the physical position of a single particle in space during particle tracking. The higher the positioning accuracy (reflected by a lower positioning accuracy value in length units), the more accurately the particle is located.

[0076] Temporal resolution refers to the exposure time when imaging particles using an imaging instrument. In other words, in particle tracking, how fast the shutter speed can be used to track moving particles. The shorter the exposure time, the higher the temporal resolution. The exposure time determines the number of photons collected by the imaging instrument. The shorter the exposure time, the fewer photons the imaging instrument collects.

[0077] In particle tracking technology, positioning accuracy and tracking time resolution are important metrics. These two indicators determine the performance of the technology. The higher the positioning accuracy and the higher the tracking time resolution, the higher the performance of the particle tracking technology. In particle tracking technology, positioning accuracy and tracking time resolution depend on the strength of the particle's optical signal.

[0078] In an imaging system dominated by shot noise, localization accuracy is inversely proportional to the square root of the number of photons collected by the imaging instrument. Therefore, temporal resolution constrains localization accuracy: the higher the temporal resolution (the shorter the exposure time), the fewer photons reach the detector, and the lower the localization accuracy. Therefore, localization accuracy is closely related to temporal resolution, and different temporal resolutions produce different localization accuracies. When referring to localization accuracy, we generally refer to the localization accuracy at a specific temporal resolution. For example, a localization accuracy of 1 nanometer at 1 millisecond means that when an image is taken with a 1 millisecond exposure time, the particle's localization accuracy is 1 nanometer. In particle tracking applications, a key performance metric is "localization accuracy at a specific temporal resolution." High performance means achieving high localization accuracy at higher temporal resolutions.

[0079] Conventional experimental methods for determining positioning accuracy can be found in Ando J, Nakamura A, Visootsat A, et al. "Single-nanoparticle tracking with angstrom localization precision and microsecond time resolution". Biophys. J., 2018, 115(12): 2413-2427. Simply put, it is first necessary to take a picture of a fixed (non-moving) particle at a specific time resolution (for example, take 5000 pictures). Then, image processing is used to obtain the position coordinates of the particle in each picture. Then, all the position coordinates (such as 5000) are uniformly plotted in a coordinate system to obtain a scatter plot of the position of the particle. The position standard deviation of the scatter plot is the positioning accuracy.

[0080] For particle tracking technology, high temporal resolution can track faster biochemical processes (such as rapid protein folding, enzyme catalysis, spatial conformational changes of nucleic acid aptamers, virus infection of cells, nanoparticle entry into cells, antigen-antibody interactions, etc.), or track faster-moving particles (such as smaller particles, high-speed moving bacteria and viruses, etc.); high positioning accuracy can monitor biochemical processes with smaller spatial changes (such as conformational changes during protein folding, the movement of protein nucleic acids or viruses in cells, the production and secretion of vesicles, etc.).

[0081] Example 4-1 Relationship between the diameter of the microlens and the detection particle tracking positioning accuracy

[0082] Microlenses of 22 μm, 34 μm, 52 μm, 73 μm, 103 μm, 350 μm, and 400 μm (the refractive index of the lens selected in this embodiment is 2.2) were selected to track particles in a solution (gold particles with a diameter of 100 nm) with a time resolution of 1 millisecond. The positioning accuracy of the particles under microlenses of different diameters was determined.

[0083] The results are shown in Table 1 and Figure 7. Figure 7 shows the positioning accuracy of particles at a time resolution of 1 millisecond under microlenses of different diameters. It can be seen that the positioning accuracy value first decreases (i.e., positioning becomes more accurate) and then increases (i.e., positioning becomes less accurate) with the diameter of the microlens. Specifically, within the microlens diameter range of 30-400 microns, the positioning accuracy is less than 4.2 nanometers; within the microlens diameter range of 35-350 microns, the positioning accuracy is less than 3.9 nanometers; within the microlens diameter range of 40-300 microns, the positioning accuracy is less than 3.5 nanometers; and within the microlens diameter range of 60-120 microns, the positioning accuracy is less than 3.0 nanometers.

[0084] It can be seen that in some embodiments of the present application, microlenses in a specific diameter range can achieve higher positioning accuracy, thereby being able to more effectively monitor biochemical processes with smaller spatial variations.

[0085] Table 1. Particle positioning accuracy at a time resolution of 1 millisecond under microlenses of different diameters (refractive index 2.2)

[0086] Example 4-2 Relationship between the diameter of the microlens and the detection particle tracking positioning accuracy

[0087] Microlenses of 22 μm, 34 μm, 52 μm, 73 μm, 103 μm, 350 μm, and 400 μm (the refractive index of the lens selected in this embodiment is 1.65) were selected to track particles in a solution (gold particles with a diameter of 100 nm) with a time resolution of 1 millisecond. The positioning accuracy of the particles under microlenses of different diameters was determined.

[0088] The results are shown in Table 2 and Figure 8. Figure 8 shows the positioning accuracy of particles at a time resolution of 1 millisecond under microlenses of different diameters. It can be seen that the positioning accuracy value first decreases (i.e., positioning becomes more accurate) and then increases (i.e., positioning becomes less accurate) with the diameter of the microlens. Specifically, within the microlens diameter range of 30-400 microns, the positioning accuracy is less than 6.2 nanometers; within the microlens diameter range of 35-350 microns, the positioning accuracy is less than 5.5 nanometers; within the microlens diameter range of 40-300 microns, the positioning accuracy is less than 5.0 nanometers; and within the microlens diameter range of 60-120 microns, the positioning accuracy is less than 4.8 nanometers.

[0089] It can be seen that in some embodiments of the present application, microlenses in a specific diameter range can achieve higher positioning accuracy, thereby being able to more effectively monitor biochemical processes with smaller spatial variations.

[0090] Table 2. Particle positioning accuracy at a time resolution of 1 millisecond under microlenses of different diameters (refractive index 1.65)

[0091] Example 4-3 Relationship between the diameter of the microlens and the tracking time resolution

[0092] Select microlenses of 22 μm, 34 μm, 52 μm, 73 μm, 103 μm, 350 μm, and 400 μm (the refractive index of the lens used in this example is 2.2) to track particles in a solution (using gold particles with a diameter of 100 nm as an example). The positioning accuracy is required to be less than 4 nm. Determine the temporal resolution that meets the required positioning accuracy for microlenses of different diameters.

[0093] The results are shown in Table 3 and Figure 9. Figure 9 shows the temporal resolution for microlenses of different diameters, when the particle positioning accuracy is required to be less than 4 nanometers. It can be seen that the temporal resolution value first decreases (i.e., the temporal resolution becomes finer) and then increases (i.e., the temporal resolution becomes less fine) with increasing microlens diameter. Specifically, for microlens diameters ranging from 30 to 400 microns, the temporal resolution is less than 1.00 milliseconds; for microlens diameters ranging from 35 to 350 microns, the temporal resolution is less than 0.90 seconds; for microlens diameters ranging from 40 to 300 microns, the temporal resolution is less than 0.80 milliseconds; and for microlens diameters ranging from 60 to 120 microns, the temporal resolution is less than 0.50 milliseconds.

[0094] It can be seen that in some embodiments of the present application, microlenses in a specific diameter range can achieve higher temporal resolution, thereby being able to more effectively monitor faster biochemical processes or track faster moving particles.

[0095] Table 3 Time resolution when particle positioning accuracy is less than 4 nm under microlenses of different diameters (refractive index is 2.2)

[0096] Example 4-4 Relationship between the diameter of the microlens and the tracking time resolution

[0097] Select microlenses of 22 μm, 34 μm, 52 μm, 73 μm, 103 μm, 350 μm, and 400 μm (the refractive index of the lens selected in this example is 1.65) to track particles in a solution (using gold particles with a diameter of 100 nm as an example). The positioning accuracy is required to be less than 4 nm. Determine the temporal resolution that meets the positioning accuracy requirement for microlenses of different diameters.

[0098] The results are shown in Table 4 and Figure 10. Figure 10 shows the temporal resolution for microlenses of different diameters, when the particle positioning accuracy is required to be less than 4 nanometers. It can be seen that the temporal resolution value first decreases (i.e., the temporal resolution becomes finer) and then increases (i.e., the temporal resolution becomes less fine) with increasing microlens diameter. Specifically, for microlens diameters ranging from 30 to 400 microns, the temporal resolution is less than 3.53 milliseconds; for microlens diameters ranging from 35 to 350 microns, the temporal resolution is less than 2.53 seconds; for microlens diameters ranging from 40 to 300 microns, the temporal resolution is less than 2.38 milliseconds; and for microlens diameters ranging from 60 to 120 microns, the temporal resolution is less than 2.25 milliseconds.

[0099] It can be seen that in some embodiments of the present application, microlenses in a specific diameter range can achieve higher temporal resolution, thereby being able to more effectively monitor faster biochemical processes or track faster moving particles.

[0100] Table 4 Time resolution when particle positioning accuracy is less than 4 nm under microlenses of different diameters (refractive index is 1.65)

[0101] It should be noted that the several embodiments provided herein are not limited to the data provided in the aforementioned embodiments. Any data within the disclosed range of this application falls within the scope of protection of this application. For example, the microlenses provided in the aforementioned embodiments have refractive indices of 2.2 and 1.65, but these are not limited to these refractive indices. In practice, the technical solutions and corresponding technical effects of this application can be achieved as long as they meet the ranges provided in this application.

[0102] The above-mentioned embodiments are only preferred embodiments of the present application and cannot be used to limit the scope of protection of the present application. Any non-substantial changes and replacements made by technicians in this field based on the present application shall fall within the scope of protection required by the present application.

[0103] It can be understood that the various technical features of the above-described embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the various technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0104] The above are merely preferred embodiments of the present application and only specifically describe the technical principles of the present application. These descriptions are intended only to explain the principles of the present application and should not be construed in any way as limiting the scope of protection of the present application. Based on the explanations herein, any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present application, as well as other specific implementations of the present application that can be conceived by those skilled in the art without inventive effort, shall be included within the scope of protection of the present application.

Claims

1. A particle tracking device, characterized in that: include: Micro lens chip, Wherein, the microlens chip includes a plurality of microlenses, and the diameter of the microlenses is 30-400 microns.

2. A particle tracking method, characterized in that: include: performing particle tracking by means of a microlens chip, in particular performing particle tracking by means of a particle tracking device comprising said microlens chip, Wherein, the microlens chip includes a plurality of microlenses, and the diameter of the microlenses is 30-400 microns.

3. Use of a microlens chip for particle tracking, in particular use of a microlens chip for particle tracking by a particle tracking device, or in particular use of a microlens chip for particle tracking by a particle tracking method, more particularly use of a microlens chip for particle tracking by a particle tracking device with a particle tracking method, characterized in that The microlens chip includes a plurality of microlenses, and the diameter of the microlenses is 30-400 microns.

4. A method for preparing a particle tracking device, characterized in that: The method comprises preparing a microlens chip, Wherein, the microlens chip includes a plurality of microlenses, and the diameter of the microlenses is 30-400 microns.

5. The device, method, or use according to any one of claims 1 to 4, characterized in that: The diameter of the microlens is 35-350 micrometers, in particular 40-300 micrometers, in particular 45-250 micrometers, in particular 50-200 micrometers, in particular 55-150 micrometers, and more in particular 60-120 micrometers.

6. The device, method, or use according to any one of claims 1 to 5, characterized in that: The microlens has a particle positioning accuracy of less than or equal to 4.0 nanometers, in particular less than or equal to 3.7 nanometers, in particular less than or equal to 3.5 nanometers, in particular less than or equal to 3.0 nanometers at a time resolution of 1 millisecond; and / or The time resolution of the microlens when the particle positioning accuracy is required to be less than 4 nanometers is less than or equal to 1.00 milliseconds, in particular less than or equal to 0.90 seconds, in particular less than or equal to 0.80 milliseconds, in particular less than or equal to 0.50 milliseconds.

7. The device, method, or use according to any one of claims 1 to 6, characterized in that: The microlens chip further comprises: a substrate, wherein the microlenses are disposed on the substrate; and a coating layer disposed on the substrate and surrounding the microlenses; or The method for preparing the microlens chip comprises: Setting the base; Disposing the microlens on the substrate; and Disposing a coating layer on the substrate so that the coating layer surrounds the microlens; Optionally, the substrate is a transparent substrate, in particular a transparent glass substrate; Optionally, each of the microlenses is arranged in a hexagonal stack on a plane above the substrate; Optionally, the coating layer comprises a polymer material, in particular a polymer resin material, more particularly polydimethylsiloxane or epoxy resin, more particularly polydimethylsiloxane, or consists of the same.

8. The device, method, or use according to any one of claims 1 to 7, characterized in that: The microlens comprises a dielectric material, in particular barium titanate, or consists of the same; Optionally, the microlens is a spherical microlens; Optionally, the microlens is a spherical microlens; Optionally, the microlens is a hemispherical microlens; Optionally, the microlens is a cylindrical lens; Optionally, the microlens is an aspherical microlens, in particular an ellipsoidal microlens, a parabolic microlens or a hyperbolic microlens; Optionally, the refractive index of the microlens is 1.65-2.20, more particularly 1.80-2.10, more particularly 1.90-2.00, more particularly 1.91-1.93, more particularly 1.92; Optionally, in the microlens chip, the number of the microlenses is greater than or equal to 1, in particular greater than or equal to 10, in particular greater than or equal to 100, in particular greater than or equal to 10 2 to 10, more particularly 10 3 to 10; Optionally, the microlenses are arranged on a plane in hexagonal stacking.

9. The device, method, or use according to any one of claims 1 to 8, characterized in that: The particle tracking device further includes a liquid channel, wherein the liquid channel includes: a fluid reservoir adjacent to the microlens chip, in particular adjacent to the microlens; and at least one, in particular at least two, liquid inlet and outlet, each of which is connected to the liquid storage tank, Wherein, the liquid channel and the microlens are optically connected; Optionally, the liquid channel is bonded to the microlens chip, in particular to the microlens and coating layer of the microlens chip; and / or The particle tracking device further comprises an optical device, wherein the optical device comprises an objective lens; Optionally, the optical device further comprises an optical path, a light source, and an optical detector, wherein the optical path optically connects the objective lens, the light source, and the optical detector; Optionally, the optical detector is electrically connected to a data processing device, and the data processing device is used to process data received by the optical detector; Optionally, the optical device further comprises a moving device, wherein the moving device is connected to the objective lens or the microlens chip and is used to move the objective lens or the microlens chip to adjust the distance between the objective lens and the microlens chip; Optionally, the light source is an LED light source, a halogen light source, a laser light source, or a xenon light source; Optionally, the optical path comprises a filter.

10. The device, method, or use according to any one of claims 1 to 9, characterized in that: The particles are cells, nanoparticles, liposomes, exosomes, bacteria, viruses, molecules, quantum dots, organelles, and / or nucleic acids, and in particular, the particles absorb light and emit scattered light, quantum dot luminescence, and / or fluorescence; and / or The particles have an average particle size of 1-1000 nm, in particular 10-500 nm, in particular 20-200 nm, in particular 50-150 nm, in particular 75-125 nm, in particular 90-100 nm, in particular 100 nm; and / or The particles are located in the liquid reservoir at a distance from the microlens greater than or equal to 0 μm, particularly greater than or equal to 1 μm, particularly greater than or equal to 2 μm, particularly greater than or equal to 3 μm, particularly greater than or equal to 4 μm, particularly greater than or equal to 4.5 μm, particularly greater than or equal to 5 μm, or particularly 0-10 μm, particularly 0-5 μm, particularly 0-3 μm, particularly 0-1 μm.

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