A kind of all-dielectric optical flow control virus sorting chip system and virus sorting method based on optical flow control technology

Abstract

The present application relates to a kind of all-dielectric optical flow control virus sorting chip system and virus sorting method based on optical flow control technology.The all-dielectric optical flow control virus sorting chip system is composed of microfluidic chip and laser control module, the microfluidic chip is multilayer bonding structure, including substrate layer (1) sequentially arranged from bottom to top, lossless high refractive layer (2), metasurface layer (3), microchannel layer (4) and Quartz quartz layer (5), two PDMS rectangles (6) are arranged on Quartz quartz layer (5) with interval;The laser control module is arranged above microfluidic chip, and provides accurate optical field excitation for microfluidic chip.The all-dielectric optical flow control virus sorting chip system provided by the present application binds laser by BIC theory, the single virus capture control efficiency is more than 95% under the condition of 5 μL / hour high throughput of chip, can be quantified purification, adapt fluorescent antibody, help window period sorting, also promote single virus research, and the flux is large, and universality is strong.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biomedical and high-performance photonic devices, and in particular to a full-dielectric optofluidic virus sorting chip system and a virus sorting method based on the optical fluidic technology. BACKGROUND

[0002] The global spread of viruses highlights the urgent need for high-sensitivity and high-specificity detection technology. Although the polymerase chain reaction (PCR) technology is still the "gold standard" for clinical diagnosis, the PCR method uses reverse transcription and real-time polymerase chain reaction (RT-PCR) to amplify the nucleic acid of the pathogen, while the amplification product is detected in real time by a fluorescent probe. The PCR method has the characteristics of strong specificity and high accuracy, and is mature in technology and widely used by society. For example, patent CN112159868A provides a novel coronavirus fluorescent qRT-PCR rapid detection system, which optimizes the primer probe targeting the novel coronavirus ORF1ab gene, N gene, and the enzyme mixture (reverse transcriptase, DNA polymerase, and UNG enzyme) of the amplification system, magnesium ion concentration, etc., to optimize the amplification system and the amplification program, thereby shortening the amplification time to within 30 minutes, greatly shortening the detection time, while ensuring the sensitivity, specificity, repeatability, and accuracy of the novel coronavirus detection. Patent CN107058622A provides a kit for combined detection of respiratory pathogens by multiplex fluorescent PCR, which contains six components: reaction liquid A, reaction liquid B, reaction liquid C, enzyme mixture, positive control, and negative control, which contains 11 common respiratory pathogens detection (influenza A virus universal type, influenza B virus, respiratory syncytial virus, human parainfluenza virus 1 / 2 / 3 type, adenovirus, Mycoplasma pneumoniae, Chlamydia pneumoniae, Legionella pneumophila, Streptococcus pneumoniae, Haemophilus influenzae, and Group A Streptococcus), which is amplified by three reaction buffers, each reaction buffer contains four fluorescence channels, which can detect 90% of the pathogen infections in clinical practice. However, the PCR method has relatively high requirements for operators and equipment, and the detection time is usually several hours. Most importantly, the fluorescent quantitative PCR method can only detect the minimum virus concentration corresponding to the cycle number (Ct value) at which the fluorescence reaches the pre-set threshold value. If the virus concentration is less than the threshold value, it cannot be detected, and the amount of virus contained in the window period patient is usually small, resulting in false negatives. Therefore, there is an urgent need for a technology that can rapidly capture and isolate any single virus in the early stages of viral infection.

[0003] At present, there is no simple and effective method and structure that can achieve single virus sorting and manipulation in a short time. SUMMARY

[0004] The application aims to overcome the defects of the prior art, and provides a full-dielectric optical flow control virus sorting chip system and a virus sorting method based on optical flow control technology.

[0005] The application can achieve the above-mentioned purposes.

[0006] The application provides a full-dielectric optical flow control virus sorting chip system, which is composed of a microfluidic chip and a laser control module.

[0007] The substrate layer is a cuboid structure with a thickness of 700-800 mu m, and the material is Si.

[0008] The laser control module excites the quasi-BIC mode of the nanocavity, generates optical gradient force, and realizes sorting of viruses in the fluid to be sorted when flowing through the nanocavity array.

[0009] Further, the laser control module provides precise optical field excitation for the microfluidic chip, which is the key to exciting the BIC mode and generating enhanced optical force. -2 The laser wavelength of the laser control module is 532 nm, which is consistent with the resonant wavelength of the nanocavity array, so as to ensure the excitation of the quasi-BIC point.

[0010] Further, the fluid interface is a hole with a diameter of 1-2 mm.

[0011] Further, the substrate layer is a cuboid structure with a thickness of 700-800 mu m, and the material is Si.

[0012] Further, the lossless high-refractive layer is lossless high-refractive silicon dioxide or elemental silicon with a thickness of 1-5 μm, and the thickness of the lossless high-refractive layer is preferably 2 μm; the lossless high-refractive layer serves as an intermediate medium layer to improve the light field constraint capability.

[0013] Further, the material of the metasurface layer is Si3N4, TiO2 or ZnO, and preferably Si3N4. The thickness of the metasurface layer is sub-100 nm (preferably 100 nm), and the metasurface layer serves as a core functional layer to excite BIC modes through a nano-cavity array to generate enhanced optical forces for virus capture.

[0014] The metasurface layer has a circular nano-cavity array arranged in the middle region, the nano-cavity diameter is 80-150 nm (preferably 110 nm), the nano-cavity arrangement period is 334 nm, and the reflectivity is >99% at a working wavelength of 532 nm.

[0015] Further, the thickness of the micro-channel layer is ≤1 μm; the micro-channel layer preferably has a thickness of 1 μm, and is used to construct a micro-fluid channel to guide the precise transmission of the virus sample above the metasurface layer. The micro-channel arranged on the micro-channel layer is prepared by spin-coating PDMS and then laser engraving ablation, the channel width is 20-30 μm, and preferably 20 μm, and the bonding with the metasurface layer has no leakage.

[0016] Further, the Quartz layer has a cuboid structure, is used to cover and protect the underlying structure, has high optical transparency, and does not affect laser incidence and fluorescence signal reading; the light transmission wavelength band of the Quartz layer covers 240-1100 nm, and is suitable for 532 nm laser and fluorescence sorting.

[0017] Further, the PDMS rectangular block is used for top fixation and fluid passage extension to ensure the sealing of the chip interface with the external hose, one small hole is drilled on each surface of the two PDMS rectangular blocks, and the holes correspond to the holes of the lossless high-refractive layer for fixing the hose.

[0018] The technical scheme 2 of the present application provides a virus sorting method based on optical flow control technology, which comprises the following steps:

[0019] S1. Sample pretreatment

[0020] The virus sample to be sorted is taken, impurity particles are removed through a 0.22 μm filter membrane, and a virus sample solution is obtained after dilution with physiological saline (preferably at a ratio of 1:1) to serve as a fluid to be sorted;

[0021] The virus sample to be sorted is taken from, for example, saliva, blood diluent, dairy virus extraction liquid, etc.

[0022] The concentration of the virus sample solution is 10 2 -106 CFU / mL;

[0023] S2. Device parameter initialization

[0024] Fluid parameter setting: inject the fluid to be sorted obtained in S1 into the fluid interface on one side of the microfluidic chip, control the flow rate and flux of the fluid to be sorted in the microchannel of the microchannel layer (4) through the external micro pump; the flow rate is generally about 100 pm / s, and the corresponding flux is stabilized at 5 pL / hour, which matches the light field confinement range of the nanocavity array of the metasurface layer (3), so as to avoid the virus from flowing out without being captured due to too fast flow rate; when other size chips are selected, the flux can be changed by changing the flow rate;

[0025] Laser parameter setting: turn on the laser control module (532 nm), select the power mode according to the sorting requirement, set the single virus sorting mode to 10-80 mW, and set the large amount of virus purification sorting mode to 80-100 mW, and the laser spot is aligned with the nanocavity array region of the chip metasurface layer (3) (ensure that the light field covers the nanocavity and excites the enhanced optical force);

[0026] S3. Virus precise sorting operation

[0027] Virus capture: when the fluid to be sorted flows through the metasurface layer, the quasi-BIC mode of the nanocavity array is excited by the laser to generate an optical gradient force, and the virus is captured in the nanocavity;

[0028] Directional movement sorting: the laser position is finely adjusted (accuracy ≤1 pm) through the laser control module to drive the captured virus to move along the PDMS microchannel direction, at this time, the virus is sorted into the nanocavity array (virus collection area, which corresponds to the transparent observation area below the Quartz layer, facilitating subsequent sorting) arranged on the chip; after a period of capture, the laser is turned off, and the impurities not captured flow out of the chip with the fluid; the laser is turned on again, and the virus capture state is maintained, and the separation and purification of the virus and the impurities are completed.

[0029] In some specific embodiments, when the virus sample concentration >10 5 CFU / mL, the laser power is increased to 80-100 mW to expand the light field coverage range (such as covering ≥100 nanocavities), so that a large amount of viruses are simultaneously captured in the nanocavity array; after 10 minutes of capture, the laser is turned off for 10 seconds, and the impurities not captured flow out of the chip with the fluid; the laser is turned on again (50 mW), and the virus capture state is maintained, and the separation and purification of the virus and the impurities are completed.

[0030] Further, the application provides a virus detection method, which adds a virus detection reagent to the fluid to be detected on the basis of the above sorting, and judges the virus sorting result through the reagent reaction result. Further, the application provides a virus detection method, which adds a virus detection reagent to the fluid to be detected on the basis of the above sorting, and judges the virus sorting result through the reagent reaction result.

[0031] For example, using fluorescent stains, the higher the concentration of viruses sorted, the higher the fluorescence in the chip.

[0032] The application of the technical solution 3 of the present application provides a full-dielectric optical flow control virus sorting chip system and a virus sorting method based on optical flow control technology, and the application includes: sorting and detection of window period single virus, screening of virus inhibitors, single virus mechanism research or virus sample purification.

[0033] Compared with the prior art, the present application has the following beneficial effects:

[0034] The present application creates a full-dielectric non-absorbing two-dimensional nanocavity array, and applies the continuous domain bound state (BIC) theory to bind the laser beam in the nanocavity to form a large range of discrete bound fields. This technology can exert enhanced optical force on the fast-moving viruses in the microchannel to achieve efficient capture of viruses. The developed chip can realize multifunctional precise capture and manipulation of any single virus under the condition of high throughput of 5 microliters / hour, with a capture efficiency of more than 95%, and also has the functions of quantification and purification. By optimizing the stability of viruses in the microflow field and the contact efficiency of viruses with the flow field, it is ensured that the captured viruses can fully combine with the subsequently introduced fluorescent staining antibodies, thereby laying a foundation for window period single virus sorting and detection. Compared with the existing virus sorting and detection technology, the present application has the advantages of large sorting and detection throughput and strong universality. In addition, this technology can also promote the development of single virus analysis methods, including virus inhibitor screening, single virus mechanism research, and drug testing and other application fields. Specifically:

[0035] 1. A new full-dielectric nanocavity array can concentrate light in a hole array with a diameter of about 100 nanometers. No absorption of light can avoid damage to biological samples due to temperature rise.

[0036] 2. Integrated optical flow control chip system. The channel is less than 1 micrometer, which can realize 99% virus capture.

[0037] 3. Efficient multifunctional manipulation of single virus and large number of viruses.

[0038] 4. Fluorescent antibody sorting and detection of viruses with single virus precision, with a sorting and detection time of less than 2 hours. BRIEF DESCRIPTION OF DRAWINGS

[0039] Figure 1 The optical flow control chip structure (a) and the schematic diagram of capturing viruses (b) of the virus sorting method of the new optical flow control technology of the present application embodiment; Figure 1 Figure 1

[0040] Figure 2 Flowchart of the method of the present application;​​

[0041] Figure 3 The band structure diagram of the single-layer metasurface structure in an embodiment of the present invention is shown below. Figure 3 a) and quality factor ( Figure 3 (b) Figure 3 In the figure, c represents the optical power of the four bands; the inset shows the electric field diagrams for the four different modes.

[0042] Figure 4 This is a parameter scan of the aperture radius in the photonic crystal plate according to an embodiment of the present invention;

[0043] Figure 5 The reflectance spectra of the single-layer metasurface structure in this invention are shown in underwater (…). Figure 5 (a) and (in the air) Figure 5 (b)

[0044] Figure 6 The resulting image of the metasurface fabricated according to an embodiment of the present invention ( Figure 6 (a) and chip physical image ( Figure 6 (b)

[0045] Figure 7 The relationship between temperature increment and laser intensity in an embodiment of the present invention ( Figure 7 (a) and the temperature distribution of the captured virus over a period of time (a) Figure 7 (b)

[0046] Figure 8 SEM images of viruses captured by the chip in this embodiment of the invention ( Figure 8 Images of (a) and various viruses captured under laser irradiation (a) Figure 8 (b)

[0047] Figure 9 The probability of a virus moving with the speed of the light beam during capture in this embodiment of the invention ( ) Figure 9 (a) and the histogram of the probability of captured adenovirus as a function of capture time and laser power. Figure 9 (b)

[0048] Figure 10 The images show fluorescent antibody tests for different concentrations of the virus in this invention (the higher the virus concentration, the brighter the fluorescence). Detailed Implementation

[0049] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. These embodiments are based on the technical solution of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments.

[0050] Embodiment 1: A full-dielectric optical flow control virus sorting chip system

[0051] In view of the limitations of the current virus sorting technology mentioned in the background art, such as limited single virus sorting capability, long sorting time, and high requirements for equipment and personnel, and in view of the limitations of the current optical tweezer manipulation technology in manipulating small biological particles such as viruses and exosomes, such as low capture efficiency and flux, single function, and obvious chip light absorption heating phenomenon, a new full-dielectric optical flow control virus sorting chip system is proposed to accurately manipulate single and large amounts of viruses at high flux.

[0052] The embodiment first provides a full-dielectric optical flow control virus sorting chip system, which comprises a microfluidic chip and a laser control module. Specifically, Figure 1 The structure diagram of the microfluidic chip and the virus capture diagram are shown.

[0053] Figure 1 The structure diagram of the microfluidic chip shown in a is shown: the microfluidic chip is a multi-layer bonding structure, including from bottom to top a substrate layer 1, a lossless high-refractive layer 2, a Si3N4 metasurface layer 3, a microchannel layer 4, a Quartz quartz layer 5, and two PDMS rectangular blocks 6.

[0054] The substrate layer 1 is 730 μm thick, made of silicon (Si), in the shape of a cuboid, serving as the base of the entire chip and providing structural support. The lossless high-refractive layer 2 is 2 μm thick, made of high-refractive silicon dioxide (SiO2), in the shape of a cuboid; its surface is machined with two holes with a diameter of 1.5 mm using an acoustic wave drill, for inserting and fixing the soft tubes, serving as the interface for fluid to enter and exit the chip. The Si3N4 super surface layer 3 is a lossless silicon nitride (Si3N4) dielectric layer 100 nm thick, in the shape of a cuboid; the middle region is arranged with an array of nanometer resonant cavities (nanocavities for short) with a diameter of 110 nm, which is the key structure for realizing the core functions of the chip (such as optical regulation, particle capture, etc.). The microchannel layer 4 is a polydimethylsiloxane (PDMS) layer 1 μm thick; it is prepared by spin-coating PDMS on the surface of SiO2 and using laser engraving to ablate the microchannel pattern, thereby preparing a microchannel pattern bonded with the Si3N4 super surface, for guiding the transmission of fluid within the chip; the fluid in the microchannel adopts a low-refractive material H2O. The Quartz quartz layer 5 is in the shape of a cuboid, serving to cover and protect the underlying structure; at the same time, due to the optical transparency of quartz, it can adapt to optical sorting or regulation requirements. Two PDMS cuboids 6 are bonded on the top of the chip; the surface of each cuboid is also drilled with two small holes, corresponding to the holes on the lossless high-refractive layer 2, for fixing or connecting external soft tubes, ensuring the integrity of the fluid passage. The layers are integrated through bonding and other methods, forming a complete microfluidic system. The two soft tubes in the figure are used for the entry and exit of fluid, and the fluid contains the material to be sorted. The all-dielectric optofluidic virus sorting chip system further comprises a laser regulation module, which provides precise optical field excitation for the microfluidic chip.

[0055] Figure 1 Figure b shows that the Si3N4 nanocavity array (on the Si3N4 super surface layer 3) with a diameter of 110 nm, under the irradiation of a large-size Gaussian light beam (wavelength: 532 nm; diameter: 10-12 μm), excites the trap mode in the dielectric nanocavity and captures viruses.

[0056] Example 2: Virus sorting method based on optical fluid control technology

[0057] Based on the all-dielectric optofluidic virus sorting chip system described above, this embodiment provides a virus sorting method based on optical fluid control technology, which realizes multifunctional virus manipulation through the design and optimization of nanometer resonant cavities, the study of virus particle dynamics in optical fluid control technology, and the design of optical fluid control chip and manipulation platform.

[0058] The virus sorting method (belonging to the optical tweezer manipulation technology) can generate light force that is tens to hundreds of times stronger than other resonance modes, thereby successfully capturing a single virus at a certain speed (greater than 100 microns per second) with a capture efficiency greater than 95%, realizing multifunctional manipulation of a single virus at a high throughput (5 microliters per hour), including capturing, transporting and positioning, and isolation and purification of a large number of viruses in the nanocavity.

[0059] Specifically, as shown in the virus sorting method includes the following steps: Figure 2

[0060] S1, design and optimization of the full-dielectric nanocavity array:

[0061] According to the working wavelength of the microfluidic chip, the period of the nanocavity array unit structure and the aspect ratio of the nanocavity array unit structure are scanned, and a full-dielectric nanocavity array with high reflectivity is designed; the intrinsic mode of the nanocavity array is analyzed, and the quasi-BIC point much larger than the quality factor of the traditional structure is found.

[0062] The super surface layer 3 in step S1 is a lossless dielectric material Si3N4, the nanocavity array unit structure is a Si3N4 circular nanocavity, and the reflectivity of the single-layer super structure at the working wavelength is greater than 99%.

[0063] S2, dynamics of virus particles in a two-dimensional potential well and an optical flow control system:

[0064] By calculating the force of each point of the virus particles in the two-dimensional light field at a specific depth, the minimum capture size of the virus particles is analyzed; at the same time, the stability of the virus in the flow field is analyzed to determine the stability of the particles and the effect of the exchange with the material in the flow field.

[0065] According to the working wavelength (532nm-550nm) of the super surface layer 3 surface structure, a two-dimensional nanocavity array of a high-reflectivity three-layer composite structure (substrate layer 1, lossless high-refractive layer 2, Si3N4 super surface layer 3) is constructed.

[0066] The nanocavity size processed in step S2 is similar to the virus, between 80-150 nanometers, and the thickness is the thickness of the silicon nitride layer, less than 100 nanometers.

[0067] S3, precise multifunctional manipulation of a single virus at a high throughput:

[0068] By adjusting the laser energy and position, any single virus can be moved to any position for precise positioning; at the same time, when the virus concentration is high, the laser energy and range are increased, a large number of viruses in the microchannel are captured within a certain time, and the single virus is isolated into a single hole, realizing purification of the virus.

[0069] ​At a higher flux (5 microliters / hour), any single virus in the microchannel is captured, and the capture efficiency is more than 95%.

[0070] The virus flow rate in the resonant cavity in step S3 can be more than 100 microns / second.

[0071] The embodiment is mainly based on the following technical principles:

[0072] The BIC mode usually relies on the interference of two optical modes, which has a certain similarity with the principle of the ring dipole. Studying the BIC mode in the nanostructure helps to explore the nature of the interaction between light and matter, and has great potential application in the fields of nano-laser, nonlinear optics, sensing and quantum optics. The present application will analyze the intrinsic mode in the nano-cavity array, find out the perfect BIC point with almost infinite quality factor and the quasi-BIC point with much larger quality factor than the traditional structure. The optical force in the perfect BIC point and the quasi-BIC point is calculated by the Minkowski stress tensor method, and the point with the most suitable optical force for optical manipulation is found out, which provides theoretical support for optical manipulation at a higher flux.

[0073] Exploring the capture stability of a single virus in the optical potential well and the faster flow field is the premise of realizing high-efficiency manipulation. The virus in the microchannel will not only be affected by the optical potential well, but also by the high-speed fluid force, so the required capture optical force is much larger than that required in static state. Therefore, the optical force and the fluid force on the particle need to be superimposed, and the capture stability under the combined force is analyzed. When the particle is captured into a single nano-cavity, the contact efficiency of the particle and the fluid without optical force and the stability of the particle in the hole also need to be analyzed. This requires simulation of the flow field near the particle and the dynamics of the particle under the restriction of the flow field and the hole.

[0074] Example 3 Verification of chip system and virus sorting method

[0075] The embodiment also verifies the above-mentioned all-dielectric optical flow control virus sorting chip system and the virus sorting method based on the optical flow control technology, and realizes the virus sorting of the new optical flow control technology.

[0076] Figure 3 The mode field analysis and optical force of the single-layer super surface structure are shown. The radius of the circular hole structure is 87 nm, the height is 100 nm, the period of the super surface is 334 nm, and the reflectivity of the super surface is 1 at a wavelength of 532 nm. Figure 3 Fig. 1, 2, 3, 4 in the figure) shows the mode energy band diagram of the TE polarized field when the hole radius and the period are 87 nm and 334 nm respectively. The quality factor of mode 3 and mode 4 at point Γ is as high as 109 However, the optical forces of both modes are small compared to other modes because the light is completely confined in the cavity with negligible leakage, hindering the interaction of light and particles. The quality factor of mode 1 is <10 3 However, the optical forces of both modes are small compared to other modes because the light is completely confined in the cavity with negligible leakage, hindering the interaction of light and particles. The quality factor of mode 1 is <10 Figure 3 b). This is because the perfect BIC mode can trap light without leakage, while mode 1 achieves a balance between beam confinement and leakage, thus realizing strong optical gradient forces (c). Figure 3 b). This is because the perfect BIC mode can trap light without leakage, while mode 1 achieves a balance between beam confinement and leakage, thus realizing strong optical gradient forces (c).

[0077] Figure 4 The parameter scanning of the small hole radius in the photonic crystal plate is shown, first scanning the small hole period 330-350nm. From the simulation results, it can be seen that when the small hole period is 334nm and 343nm, the calculated quasi-BIC is near 564THz (about 532nm). This is very consistent with the required incident light wavelength. At the same time, combined with the parameter scanning results of the small hole depth. Finally, this embodiment determines that the small hole period is 334nm and the depth is 100nm.

[0078] Figure 5 After scanning the period of the unit structure and the aspect ratio of the unit structure, the present application designs a high-reflection all-dielectric nanocavity array. Figure 5 a) it can be seen that the reflectivity of the metasurface structure of the present application is as high as 99% under water at 563.9Thz (532nm). However, there is no obvious resonance peak in air (b). Figure 5 a) it can be seen that the reflectivity of the metasurface structure of the present application is as high as 99% under water at 563.9Thz (532nm). However, there is no obvious resonance peak in air (b).

[0079] Figure 6 a) shows the silicon nitride metasurface structure prepared by the photolithography process. The packaging of the PDMS optofluidic chip usually uses the bonding of PDMS and glass, and at the same time, in order to insert the catheter later, the PDMS should also be drilled, and finally the catheter is inserted to complete the preparation of the optofluidic chip. The sample of the optofluidic chip is shown in Figure 6 b), the width of the channel of the optofluidic chip is about 20μm, and the thickness between the channels is about 1μm.

[0080] Figure 7 a) shows that when irradiated with a laser intensity of 1mW·μm -2 Figure 7 ​The middle b shows that the morphology and activity of the virus in the nanocavity remain stable as time goes on. Therefore, the all-dielectric nanocavity array of the application can manipulate the virus without heating the virus and changing the activity of the virus.

[0081] Figure 8 The middle a shows the SEM imaging results of the adenovirus in the all-dielectric nanocavity array. The bottom particles show dark zone characteristics due to light field attenuation, and the particles close to the upper surface of the hole wall show bright zones due to scattering enhancement. It is worth noting that when the particle size matches the cavity, the phenomenon of single-hole capturing a single particle is dominant, which is caused by the disturbance effect of the captured particles on the local light field. This disturbance significantly weakens the light force effect in the adjacent area, resulting in a decrease in the overall capture efficiency of the nanocavity array. Figure 8 The middle b shows that by using continuous irradiation of the nanocavity array with high laser power (60-100 mW, equivalent laser intensity ≈1 mW·μm -2 ) can achieve directional enrichment of hundreds to thousands of virus particles. As the laser power increases, the number of local points with strong optical gradient force in the nanocavity array increases significantly, thereby realizing large-scale virus capture. When the laser power reaches 100 mW, dozens of adenoviruses can be successfully captured in the light spot area.

[0082] Figure 9 The middle a shows that the probability of virus entering the nanocavity P 捕获 ) depends on the capture time. In principle, the longer the capture time, the higher the probability of the virus being captured in the nanocavity. However, in some cases, the virus is not captured in the nanocavity for a long time. Therefore, the capture probability also depends on the speed of light beam movement when the virus is transported Figure 9 The middle a. Figure 9 The middle b shows that when the capture time is longer than 20 s (the laser beam remains stationary) and the laser power is greater than 80 mW, the adenovirus capture rate is >95%.

[0083] Figure 10 The middle b shows that when the capture time is longer than 20 s (the laser beam remains stationary) and the laser power is greater than 80 mW, the adenovirus capture rate is >95%.

[0084] The structural parameters of the superstructure surface in the application are not limited to this. For different design wavelengths, the period / radius of the unit structure can be reasonably adjusted according to the design wavelength.

[0085] The preferred embodiments of the present application have been described above in detail. It should be understood that modifications and variations to the preferred embodiments could be made by those skilled in the art in light of the teachings above without departing from the spirit of the present application. It is therefore to be understood that what is desired to be protected by letters patent is defined by the scope of the claims below and that on the basis of the teachings of the present application, obvious modifications and equivalents can be adopted by those skilled in the art in their possession of the teachings of the present application without departing from the spirit and scope of the application.

Claims

1. A total dielectric photonic fluidic virus sorting chip system, characterized in that, The microfluidic chip is composed of a laser control module, and the microfluidic chip is a multilayer bonding structure, which comprises, from bottom to top, a substrate layer (1), a lossless high-refractive layer (2), a metasurface layer (3), a microchannel layer (4) and a Quartz quartz layer (5), two PDMS rectangles (6) are arranged on the Quartz quartz layer (5) at intervals; the middle region of the metasurface layer (3) is arranged with a circular nanocavity array as a virus collection area; the microfluidic channel is arranged on the microchannel layer (4); the nanocavity array and the microfluidic channel are in communication; the laser control module is arranged above the microfluidic chip to provide precise optical field excitation for the microfluidic chip; The lossless high-refractive layer (2) is provided with a basic interface on both sides, the metasurface layer (3), the microchannel layer (4) and the Quartz quartz layer (5) are provided with a cooperative interface on both sides, and the PDMS rectangle (6) is provided with an extension interface on both sides; the basic interface, the cooperative interface and the extension interface jointly form a fluid interface, and the fluid interfaces on both sides are respectively inserted into a hose for the inlet and outlet of the fluid to be sorted; The nanocavity array is affected by the laser control module, the quasi-BIC mode of the nanocavity is excited, the optical gradient force is generated, and the virus in the fluid to be sorted is sorted when flowing through the nanocavity array due to the optical gradient force.

2. The all-dielectric optical fluidic virus sorting chip system of claim 1, wherein, The laser wavelength of the laser control module is 532 nm, the adjustable range of laser power is 10-100 mW, and the equivalent laser intensity is 1 mW μm -2 ; the fluid interface is a hole with a diameter of 1-2 mm.

3. The all-dielectric optical fluidic virus sorting chip system of claim 1, wherein, The substrate layer (1) is a cuboid structure with a thickness of 700-800μm, and is made of Si, which provides structural support as a base.

4. The all-dielectric optical fluidic virus sorting chip system of claim 1, wherein, The lossless high-refractive layer (2) is a lossless high-refractive silicon dioxide or elemental silicon with a thickness of 1-5μm, which serves as an intermediate medium layer.

5. The all-dielectric optical fluidic virus sorting chip system of claim 1, wherein, The material of the metasurface layer (3) is selected from any one of Si3N4, TiO2 or ZnO, and the thickness of the metasurface layer (3) is sub-100nm; the nanocavity diameter is 80-150nm, and the nanocavity arrangement period is 334nm.

6. The all-dielectric optical fluidic virus sorting chip system of claim 1, wherein, The thickness of the microchannel layer (4) is ≤1μm; the microchannel on the microchannel layer (4) is prepared by laser ablation after spin coating PDMS, and the channel width is 20-30μm.

7. The all-dielectric optical fluidic virus sorting chip system of claim 1, wherein, The Quartz quartz layer (5) is a cuboid structure, which is used to cover and protect the lower layer structure, has high optical transparency, and the light transmission band covers 240-1100nm.

8. The all-dielectric optical fluidic virus sorting chip system according to any one of claims 1-7, wherein, The thickness of the substrate layer (1) is 730μm, the thickness of the lossless high-refractive layer (2) is 2μm, the material of the metasurface layer (3) is Si3N4, the thickness of the metasurface layer (3) is sub-100nm, the nanocavity diameter is 110nm, the nanocavity arrangement period is 334nm, the thickness of the microchannel layer (4) is 1μm, and the microchannel width on the microchannel layer (4) is 20μm.

9. A method for virus sorting based on optofluidic technology, characterized in that, The all-dielectric optical fluidic virus sorting chip system based on claim 1, the virus sorting method is for non-disease diagnosis and treatment purposes, comprising the following steps: S1. Sample pretreatment: Take the virus sample to be sorted, filter out impurity particles through a 0.22μm filter membrane, dilute with physiological saline to obtain a virus sample solution as the fluid to be sorted. The concentration of virus in the virus sample liquid ranges from 10 2 -10 6 CFU / mL; S2. Device parameter initialization Fluid parameter setting: inject the fluid to be sorted obtained in S1 into the fluid interface on one side of the microfluidic chip, and control the flow rate and flux of the fluid to be sorted in the microchannel of the microchannel layer (4) through an external micro pump; Laser parameter setting: turn on the laser control module, select the power mode according to the sorting requirement, set the single virus sorting mode to 10-80 mW, and set the large amount of virus purification sorting mode to 80-100 mW, and the laser spot is aligned with the nanocavity array region of the super surface layer (3) of the chip; S3. Virus precise sorting Virus capture: when the fluid to be sorted flows through the super surface layer (3), the quasi-BIC mode of the nanocavity array is excited by laser to generate optical gradient force, and the virus is captured in the nanocavity; Directional movement sorting: the laser position is fine-tuned through the laser control module to drive the captured virus to move along the PDMS microchannel direction, at this time, the virus is sorted into the nanocavity array arranged on the chip; after a period of capture, the laser is turned off, and the impurities not captured flow out of the chip with the fluid; the laser is turned on again, and the virus capture state is maintained, and the separation and purification of the virus and the impurities are completed.

10. Use of a full dielectric optofluidic virus sorting chip system according to claim 1 or a method for virus sorting based on optofluidics according to claim 9, characterized in that, The application includes: window period single virus sorting, virus inhibitor screening, single virus mechanism research or virus sample purification.

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