A high-throughput miniaturized dark-field biochemical sensing system

Abstract

The invention relates to the technical field of heavy metal ion detection, and in particular to a high-throughput miniaturized dark-field biochemical sensing system. When detecting heavy metal concentration, the invention designs four parallel observation areas on a multi-channel microfluidic chip, a single plasma nanosensor is fixed in advance on a substrate of the observation area through four probe inlets, a sample to be tested enters four different areas in a reflection pool through the sample inlet via a microchannel, the four areas are modified with different ligand probes, and high-throughput detection of multiple heavy metal ions is achieved by adopting four parallel observation areas; a particle recognition module for dark-field images provided by the invention adopts a scheme of copying and tiling a matrix of similar size into a matrix of image size to form a new mask, and then covering the original dark-field image with the new mask, and processing the dark-field image by effectively distinguishing particle center points, thereby improving the accuracy of particle recognition in the dark-field image.

Description

Technical Field

[0001] The invention relates to the technical field of heavy metal ion detection, and in particular to a high-throughput miniaturized dark-field biochemical sensing system. Background Art

[0002] Heavy metal pollution in water bodies refers to the pollution caused by pollutants containing heavy metal ions (such as mercury, lead, copper, cadmium, zinc, etc.) directly entering the water body without treatment. Localized surface plasmon resonance scattering spectroscopy based on precious metal nanoparticles is a new fast and sensitive spectral analysis technology. Due to its high sensitivity and accuracy, it has been widely used in dynamic monitoring and quantitative analysis of heavy metal pollution in water bodies in recent years.

[0003] In the prior art, there are technical solutions for heavy metal ion detection using a dark-field biochemical sensing system. For example, a Chinese invention patent (CN113567398A) discloses a method for detecting lead ion concentration based on dark-field spectroscopy, which includes the following steps: (1) modifying gold nanoparticles with glutathione; (2) fixing the gold nanoparticles on a glass substrate; (3) dripping a lead-containing sample on the glass substrate and rinsing after 5 minutes; (4) dripping glutathione-modified gold nanoparticles on the glass substrate and then covering it with a coverslip; (5) detecting the red shift of the scattering spectrum of the gold nanoparticles under a dark-field microscope system, and quantifying the concentration of lead ions by the red shift of the spectrum.

[0004] However, when performing metal ion detection, the above scheme cannot achieve simultaneous high-throughput detection of multiple heavy metal concentrations on the one hand, and on the other hand, when processing dark field images, there is a problem of low image processing accuracy, which affects the detection accuracy of heavy metal ions. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a high-throughput miniaturized dark-field biochemical sensing system to improve the accuracy and high-throughput detection of heavy metal ions.

[0006] The present invention provides a high-throughput miniaturized dark-field biochemical sensing system, comprising:

[0007] Main structure of miniaturized dark field system and multi-channel microfluidic chip;

[0008] The main structure of the miniaturized dark field system includes two parts: a hardware system and an image data processing system;

[0009] The image data processing system is used to perform data processing on the dark field image, so as to obtain the ion spectrum offset;

[0010] The image data processing system comprises:

[0011] Standard image acquisition module, used to collect dark-field spectra of samples without LSPR characteristics in the same environment to obtain standardized images;

[0012] A dark field image preprocessing module, used for performing denoising on the dark field image;

[0013] A dark field image particle brightness enhancement module is used to adjust and enhance the brightness of particles in the dark field image to improve the accuracy of particle recognition;

[0014] The dark field image particle brightness enhancement module uses an image mask to cover the original dark field image to maximize the brightness intensity of the particles. The mask is set to a matrix of similar size according to the pixel area occupied by the center area of ​​the particle to improve the accuracy of particle recognition.

[0015] The mask rule is: copying and tiling the similar size matrix into an image size matrix to form a new mask, and then overlaying the new mask on the original dark field image to effectively distinguish the center points of the particles;

[0016] A spectrum fitting module, used for obtaining the peak value and half-height width of the particle by using Lorentz fitting;

[0017] The heavy metal ion concentration determination module is used to calculate the heavy metal ion concentration according to the peak value and half-height width of the particles.

[0018] Preferably, the hardware system includes a dark field microscope, a dark field condenser, an intelligent terminal, a single plasma nanosensor and a modulated monochromatic light source.

[0019] Preferably, the modulated monochromatic light source is a 250W adjustable halogen lamp light source, and the white light source emitted by the halogen lamp is modulated to obtain a monochromatic incident light source with continuous wavelength modulation; the monochromatic incident light source is input into a dark field microscope, and after being focused by the dark field condenser, it is focused on the multi-channel microfluidic chip, and the scattered light after the sample to be tested in the microfluidic chip is mixed with the nanoparticles of the single plasma nanosensor can be directly observed by the naked eye through the eyepiece of the dark field microscope, and then recorded in the form of an image through the intelligent terminal to form a dark field image.

[0020] Preferably, the single plasmon nanosensor is composed of gold nanoparticles with stable chemical properties and good biocompatibility, and the single plasmon nanosensor is obtained by modifying the surface of the gold nanoparticles with specific ligand probes that can recognize heavy metal ions.

[0021] Preferably, the single plasmonic nanosensor in each observation area is modified with a different ligand probe.

[0022] Preferably, the multi-channel microfluidic chip is made of a glass substrate and a PDMS film, and four parallel observation areas are designed on the multi-channel microfluidic chip. The single plasma nanosensor is fixed in advance on the substrate of the observation area through four probe inlets. The sample to be tested enters four different areas in the reflection pool through the sample inlet via the microchannel. The four areas are modified with different ligand probes, thereby realizing the acquisition of dark field images.

[0023] The embodiments of the present invention have the following technical effects:

[0024] When detecting heavy metal concentrations, the present invention designs four parallel observation areas on a multi-channel microfluidic chip, and a single plasma nanosensor is fixed in advance on a substrate of the observation area through four probe inlets. The sample to be tested enters four different areas in a reflection pool through the sample inlet via a microchannel. The four areas are modified with different ligand probes. By adopting four parallel observation areas, high-throughput detection of multiple heavy metal ions is achieved.

[0025] At the same time, the present invention sets up a particle recognition module for dark field images, which adopts a scheme of copying and tiling the similar size matrix into an image size matrix to form a new mask, and then overlaying the new mask on the original dark field image to effectively distinguish the center points of particles to process the dark field image, thereby improving the accuracy of particle recognition in the dark field image. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0027] Figure 1 is a structural diagram of a high-throughput miniaturized dark-field biochemical sensing system provided in Example 1 of the present invention;

[0028] Figure 2 is a structural diagram of an image data processing system provided by Embodiment 1 of the present invention;

[0029] Figure 3 It is a schematic diagram of the structure of the multi-channel microfluidic chip provided in Example 1 of the present invention. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the scope of protection of the present invention.

[0031] Embodiment 1,

[0032] Figure 1 1 is a structural diagram of a high-throughput miniaturized dark-field biochemical sensing system provided in Example 1 of the present invention. Figure 1 , specifically including the following modules:

[0033] Main structure of miniaturized dark field system and multi-channel microfluidic chip;

[0034] The main structure of the miniaturized dark field system includes two parts: a hardware system and an image data processing system. The hardware system includes a dark field microscope, a dark field condenser, an intelligent terminal, a single plasma nanosensor and a modulated monochromatic light source. The modulated monochromatic light source is a 250w adjustable halogen lamp light source. The white light source emitted by the halogen lamp is modulated to obtain a monochromatic incident light source with continuous wavelength modulation. The monochromatic incident light source is input into the dark field microscope, and after being focused by the dark field condenser, it is focused on the multi-channel microfluidic chip. The scattered light after the sample to be tested in the microfluidic chip and the nanoparticles of the single plasma nanosensor are mixed can be directly observed by the naked eye through the eyepiece of the dark field microscope, and then recorded in the form of an image through the intelligent terminal to form a dark field image.

[0035] The single plasma nanosensor is composed of gold nanoparticles with stable chemical properties and good biocompatibility. The single plasma nanosensor is obtained by modifying the surface of the gold nanoparticles with specific ligand probes capable of identifying heavy metal ions.

[0036] Among them, the single plasma nanosensor in each observation area is modified with different ligand probes, so that the concentrations of multiple heavy metal ions can be identified in one test, thereby achieving high-throughput detection.

[0037] The sensing principle of the single plasma nanosensor is that under the action of the combination of heavy metal ions and specific probes, the scattering spectrum of gold nanoparticles shifts, and the concentration of heavy metal ions is quantitatively calculated based on the shift.

[0038] The image data processing system is used to perform data processing on the dark field image, so as to obtain the ion spectrum offset;

[0039] Specifically, Figure 2 As shown, the image data processing system comprises:

[0040] Standard image acquisition module, used to collect dark-field spectra of samples without LSPR characteristics in the same environment to obtain standardized images;

[0041] A dark field image preprocessing module, used for performing denoising on the dark field image;

[0042] Image denoising is an important task in the field of image processing, which aims to reduce or eliminate noise in an image to improve the quality and readability of the image. In this embodiment, a mean filter is used to perform denoising on the dark field image;

[0043] A dark field image particle brightness enhancement module is used to adjust and enhance the brightness of particles in the dark field image to improve the accuracy of particle recognition;

[0044] Among them, the dark field image particle brightness enhancement module uses an image mask to cover the original dark field image. Through the above operation, the brightness intensity of the particles can be maximized. According to the pixel area occupied by the central area of ​​the particle, the mask is set to a matrix of similar size to improve the accuracy of particle recognition;

[0045] The mask rule is: copying and tiling the similar size matrix into an image size matrix to form a new mask, and then overlaying the new mask on the original dark field image to effectively distinguish the center points of the particles;

[0046] In this embodiment, a particle recognition module for dark field images is proposed, which processes the dark field images by copying and tiling the similar size matrix into an image size matrix to form a new mask, and then overlaying the new mask on the original dark field image to effectively distinguish the center points of the particles, thereby improving the accuracy of particle recognition in the dark field images;

[0047] A spectrum fitting module, used for obtaining the peak value and half-height width of the particle by using Lorentz fitting;

[0048] A heavy metal ion concentration determination module, used to calculate the heavy metal ion concentration according to the peak value and half-height width of the particles;

[0049] As attached Figure 3 As shown, the multi-channel microfluidic chip is made of a glass substrate and a PDMS film, and 4 parallel observation areas are designed on the multi-channel microfluidic chip. The single plasma nanosensor is fixed on the substrate of the observation area in advance through 4 probe inlets. The sample to be tested enters 4 different areas in the reflection pool through the sample inlet through the microchannel, and the 4 areas are modified with different ligand probes, thereby realizing the acquisition of dark field images;

[0050] In this embodiment, high-throughput detection of heavy metal ions is achieved by adopting four parallel observation areas.

[0051] Embodiment 2, the present invention also provides an electronic device, including one or more processors and a memory. The processor may be a central processing unit (CPU) or other forms of processing units with data processing capabilities and / or instruction execution capabilities, and may control other components in the electronic device to perform desired functions.

[0052] The memory may include one or more computer program products, and the computer program product may include various forms of computer-readable storage media, such as volatile memory and / or non-volatile memory. The volatile memory may include, for example, random access memory (RAM) and / or cache memory (cache), etc. The non-volatile memory may include, for example, read-only memory (ROM), hard disk, flash memory, etc. One or more computer program instructions may be stored on the computer-readable storage medium, and the processor may run the program instructions to implement the desired functions of a high-throughput miniaturized dark-field biochemical sensing system of any embodiment of the present application described above. Various contents such as initial external parameters, thresholds, etc. may also be stored in the computer-readable storage medium.

[0053] In one example, the electronic device may further include: an input device and an output device, and these components are interconnected through a bus system and / or other forms of connection mechanisms (not shown). The input device may include, for example, a keyboard, a mouse, etc. The output device may output various information to the outside, including early warning information, braking force, etc. The output device may include, for example, a display, a speaker, a printer, a communication network and a remote output device connected thereto, etc.

[0054] Of course, for the sake of simplicity, components such as buses, input / output interfaces, etc. are omitted. In addition, the electronic device may further include any other appropriate components according to specific application conditions.

[0055] In addition to the above-mentioned methods and devices, an embodiment of the present application may also be a computer program product, which includes computer program instructions, which, when executed by a processor, enable the processor to implement the functions of a high-throughput miniaturized dark-field biochemical sensing system provided by any embodiment of the present application.

[0056] The computer program product may be written in any combination of one or more programming languages ​​to write program codes for performing the operations of the embodiments of the present application, including object-oriented programming languages, such as Java, C++, etc., and conventional procedural programming languages, such as "C" language or similar programming languages. The program code may be executed entirely on the user computing device, partially on the user device, as an independent software package, partially on the user computing device and partially on a remote computing device, or entirely on a remote computing device or server.

[0057] In addition, an embodiment of the present application may also be a computer-readable storage medium on which computer program instructions are stored. When the computer program instructions are executed by a processor, the processor implements the functions of a high-throughput miniaturized dark-field biochemical sensing system provided by any embodiment of the present application.

[0058] The computer readable storage medium can adopt any combination of one or more readable media. The readable medium can be a readable signal medium or a readable storage medium. The readable storage medium can include, for example, but is not limited to, a system, device or device of electricity, magnetism, light, electromagnetic, infrared, or semiconductor, or any combination of the above. More specific examples (non-exhaustive list) of readable storage media include: an electrical connection with one or more wires, a portable disk, a hard disk, a random access memory (RAM), a read-only memory (ROM), an erasable programmable read-only memory (EPROM or flash memory), an optical fiber, a portable compact disk read-only memory (CD-ROM), an optical storage device, a magnetic storage device, or any suitable combination of the above.

[0059] It should be noted that the terms used in the present invention are only for describing specific embodiments, rather than limiting the scope of the present application. As shown in the present specification, unless the context clearly indicates an exception, the words "one", "a", "a kind of" and / or "the" do not specifically refer to the singular, but may also include the plural. The terms "include", "comprise" or any other variant thereof are intended to cover non-exclusive inclusion, so that the process, method or device including a series of elements includes not only those elements, but also includes other elements not explicitly listed, or also includes elements inherent to such process, method or device. In the absence of more restrictions, the elements defined by the sentence "include one..." do not exclude the presence of other identical elements in the process, method or device including the elements.

[0060] It should also be noted that the terms "center", "up", "down", "left", "right", "vertical", "horizontal", "inside", "outside", etc., indicating the orientation or positional relationship, are based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. Unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", etc. should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be an indirect connection through an intermediate medium, or it can be a connection between the two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0061] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the technical solutions of the embodiments of the present invention.

Claims

1. A high-throughput miniaturized dark-field biochemical sensing system, characterized in that: include: Main structure of miniaturized dark field system and multi-channel microfluidic chip; The main structure of the miniaturized dark field system includes two parts: a hardware system and an image data processing system; The image data processing system is used to process the dark field image to obtain the ion spectrum offset; The image data processing system comprises: Standard image acquisition module, used to collect dark-field spectra of samples without LSPR characteristics in the same environment to obtain standardized images; A dark field image preprocessing module, used for performing denoising on the dark field image; A dark field image particle brightness enhancement module is used to adjust and enhance the brightness of particles in the dark field image to improve the accuracy of particle recognition; The dark field image particle brightness enhancement module uses an image mask to cover the original dark field image to maximize the brightness intensity of the particles. The mask is set to a matrix of similar size according to the pixel area occupied by the center area of ​​the particle to improve the accuracy of particle recognition. The mask rule is: copying and tiling the similar size matrix into an image size matrix to form a new mask, and then overlaying the new mask on the original dark field image to effectively distinguish the center points of the particles; A spectrum fitting module, used for obtaining the peak value and half-height width of the particle by using Lorentz fitting; The heavy metal ion concentration determination module is used to calculate the heavy metal ion concentration according to the peak value and half-height width of the particles.

2. A high-throughput miniaturized dark-field biochemical sensing system according to claim 1, characterized in that: The hardware system includes a dark field microscope, a dark field condenser, an intelligent terminal, a single plasma nanosensor and a modulated monochromatic light source.

3. A high-throughput miniaturized dark-field biochemical sensing system according to claim 2, characterized in that: The modulated monochromatic light source is a 250W adjustable halogen lamp light source, and the white light source emitted by the halogen lamp light source is modulated to obtain a monochromatic incident light source with continuous wavelength modulation; the monochromatic incident light source is input into a dark field microscope, and after being condensed by the dark field condenser, it is focused on the multi-channel microfluidic chip, and the scattered light after the sample to be tested in the microfluidic chip is mixed with the nanoparticles of the single plasma nanosensor can be directly observed by the naked eye through the eyepiece of the dark field microscope, and then recorded in the form of an image through the intelligent terminal to form a dark field image.

4. A high-throughput miniaturized dark-field biochemical sensing system according to claim 2, characterized in that: The single plasma nanosensor is composed of gold nanoparticles with stable chemical properties and good biocompatibility. The single plasma nanosensor is obtained by modifying the surface of the gold nanoparticles with specific ligand probes capable of identifying heavy metal ions.

5. A high-throughput miniaturized dark-field biochemical sensing system according to claim 2, characterized in that: The multi-channel microfluidic chip is made of a glass substrate and a PDMS film. Four parallel observation areas are designed on the multi-channel microfluidic chip. A single plasma nanosensor is fixed in advance on the substrate of the observation area through four probe inlets. The sample to be tested enters four different areas in the reflection pool through the sample inlet via a microchannel. The four areas are modified with different ligand probes, thereby realizing the collection of dark field images.

6. A high-throughput miniaturized dark-field biochemical sensing system according to claim 1, characterized in that: A single plasmonic nanosensor in each observation area is modified with a different ligand probe.

Images