Micro-nano target in-situ detection device and method

Abstract

The invention discloses a micro-nano target in-situ detection device and method, and relates to the field of detection equipment.The device comprises fluid conveying equipment, a micro-fluidic chip, image obtaining equipment and a detection device, the fluid conveying equipment is connected with the micro-fluidic chip, and the fluid conveying equipment can convey fluid at a target position into the micro-fluidic chip; the image acquisition equipment is in signal connection with the detection device, and the image acquisition equipment is used for acquiring an optical image of a micro-nano target in the fluid in the micro-fluidic chip; the detection device can perform image recognition on the optical image. The method comprises the following steps: conveying fluid at a target position into the micro-fluidic chip through fluid conveying equipment; treating the entered fluid through the micro-fluidic chip; acquiring an optical image of a micro-nano target in the fluid processed by the micro-fluidic chip through image acquisition equipment; and performing image recognition on the optical image through the detection device. In-situ treatment of an optical image direct detection technology is realized, and the method has environmental interference resistance and high instantaneity.

Description

Technical Field

[0001] The present invention relates to the technical field of detection equipment, and in particular to a device and method for in-situ detection of micro-nano targets. Background Art

[0002] The micro- and nanoscale (1nm-100μm) is the core domain of life activities, material evolution, and pollutant diffusion. To break through the boundaries of modern scientific cognition and reveal the mechanisms at this microscale, capturing the dynamic characteristics of micro- and nanoscale targets in real environments has become a key challenge. The core solution lies in developing high-precision in-situ detection technologies for micro- and nanoscale targets. Existing micro- and nanoscale target detection technologies can be mainly divided into two types: indirect detection technologies based on converted signals such as electrical and acoustic signals, and direct detection technologies based on optical images.

[0003] Indirect detection technology based on conversion signals usually has a fast response speed and has therefore been widely used in detection scenarios. However, its problem is that the conversion signal is easily interfered with by environmental factors such as temperature and humidity, resulting in poor detection accuracy and stability in complex environments. Direct detection technology based on optical images has the advantages of high detection accuracy and low sensitivity to environmental influences, but its problem is that the sample usually needs to be pre-processed and made into a sufficiently thin glass slide specimen so that the light emitted by the light source can fully penetrate the sample to ensure a clear optical image. This greatly limits its application in in-situ detection scenarios. Therefore, there is an urgent need for an in-situ detection device and method for micro-nano targets that is both resistant to environmental interference and highly immediate. Summary of the Invention

[0004] The purpose of the present invention is to provide a micro-nano target in-situ detection device and method to solve the problems existing in the above-mentioned prior art, realize the in-situ direct optical image detection technology, and have resistance to environmental interference and high immediacy.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides an in-situ detection device for micro-nano targets, comprising a fluid conveying device, a microfluidic chip, an image acquisition device, and a detection device. The fluid conveying device is connected to the microfluidic chip and can convey fluid at a target position into the microfluidic chip; the image acquisition device is used for signal connection with the detection device and is used to acquire an optical image of the micro-nano target in the fluid in the microfluidic chip; and the detection device can perform image recognition on the optical image.

[0007] Preferably, the detection device is capable of identifying at least one of the micro-nano targets, and the detection device is capable of obtaining at least two types of characteristic information of each of the micro-nano targets, and the characteristic information at least includes the category, content, contour shape, contour size, position coordinates and area of ​​each of the micro-nano targets.

[0008] Preferably, an observation platform is further included, which includes a platform body, a clamp and a light source. A mounting hole is provided on the working surface of the platform body, and the light source is embedded in the mounting hole; the clamp is connected to the platform body, and the clamp can clamp the microfluidic chip on the working surface; the light source is arranged on the side of the microfluidic chip away from the image acquisition device.

[0009] Preferably, the clamp includes two clips, two elastic components and two locking components, and the clips, the elastic components and the locking components correspond to each other one by one; each elastic component is arranged between the working surface and the corresponding clip; each clip can press the corresponding clip and the corresponding elastic component onto the working surface by being fixedly connected to the observation plane.

[0010] Preferably, each of the locking members includes a rod and a limiting member, one end of each of the rods is fixedly connected to the corresponding limiting member, and the other end of each of the rods is provided with a thread; a through hole is provided on each of the clips; each of the rods can pass through the through hole of the corresponding clip and the inner cavity of the corresponding elastic component in sequence and be fixedly connected to the platform body, and when the rod is fixedly connected to the platform body, the limiting member can press the corresponding clip and the corresponding elastic component tightly on the working surface.

[0011] Preferably, a lifting device is further included, the image acquisition device is arranged above the microfluidic chip, the observation platform is connected to the lifting device, and the lifting device can drive the observation platform to move away from the image acquisition device or towards the image acquisition device.

[0012] Preferably, it also includes a slider, and the lifting device includes a driving device, a screw and a guide rod; the observation platform is fixedly connected to the slider; the slider is threadedly connected to the screw; the slider is slidingly connected to the guide rod; the driving device can drive the screw to rotate around the axis of the screw, and the driving device can move the slider along the axial direction of the screw by driving the screw to rotate around the axis of the screw.

[0013] Preferably, the image acquisition device is a camera, and the lens of the image acquisition device is a microscope lens; the fluid delivery device is a microfluidic pump.

[0014] Preferably, an electric field applying device is further included, which can apply an electric field at the flow channel of the microfluidic chip and directional arrange the micro-nano targets in the flow channel of the microfluidic chip.

[0015] The present invention also provides a micro-nano target in-situ detection method based on the micro-nano target in-situ detection device, comprising the following steps:

[0016] S1, delivering the fluid at the target location to the microfluidic chip through the fluid delivery device;

[0017] S2. Processing the fluid entering the microfluidic chip through the microfluidic chip;

[0018] S3, acquiring, by the image acquisition device, the optical image of the micro-nano target in the fluid processed by the microfluidic chip;

[0019] S4. Performing image recognition on the optical image by the detection device.

[0020] Compared with the prior art, the present invention has achieved the following technical effects:

[0021] The present invention provides an in-situ detection device and method for micro-nano targets, comprising a fluid delivery device, a microfluidic chip, an image acquisition device, and a detection device. The fluid delivery device is connected to the microfluidic chip and is capable of delivering fluid at a target location into the microfluidic chip; the image acquisition device is used for signal connection with the detection device and is used to acquire an optical image of the micro-nano target in the fluid within the microfluidic chip; and the detection device is capable of performing image recognition on the optical image. The present invention delivers the fluid at the target location into the microfluidic chip via the fluid delivery device, uses the microfluidic chip as a sample carrier, and performs online sample processing via the microfluidic chip, thereby achieving in-situ sample collection and on-chip processing. This directly integrates the sample processing and preparation stage in traditional detection technology into the detection stage, achieving in-situ detection. Furthermore, the microfluidic chip has good light transmittance, enabling the image acquisition device to acquire an optical image of the micro-nano target and perform detection via the detection device, thereby achieving in-situ detection of optical image direct detection technology, with resistance to environmental interference and high immediacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0023] Figure 1Schematic diagram of the structure of the micro-nano target in-situ detection device provided in Example 1;

[0024] Figure 2 is a schematic structural diagram of the observation platform provided in Example 1;

[0025] Figure 3 is a schematic structural diagram of the microfluidic chip provided in Example 1;

[0026] Figure 4 is a front view of the platform body provided in Example 1;

[0027] Figure 5 yes Figure 4 A top view of

[0028] Figure 6 yes Figure 4 Right view;

[0029] Figure 7 is a front view of the light source provided in Example 1;

[0030] Figure 8 yes Figure 7 A top view of

[0031] Figure 9 is a schematic structural diagram of the top plate provided in Example 1;

[0032] Figure 10 This is a schematic diagram of the structure of the mainboard provided in Example 1;

[0033] Figure 11 is a schematic structural diagram of the base plate provided in Example 1;

[0034] Figure 12 is an enlarged view of the sample inlet and sample outlet of the microfluidic chip provided in Example 1;

[0035] Figure 13 Schematic diagram of the process of in-situ detection of micro-nano targets provided in Example 2;

[0036] Figure 14 This is the micron-scale water droplet image and detection effect diagram of Example 3;

[0037] In the figure: 100, micro-nano target in-situ detection device; 1, fluid delivery device; 2, target position; 3, microfluidic chip; 3-1, top plate; 3-2, main board; 3-3, bottom plate; 3-4, sample inlet; 3-5, pre-treatment area; 3-6, detection area; 3-7, sample outlet; 4, image acquisition device; 5, detection device; 6, waste liquid collection tank; 7, lifting device; 7-1, driving device; 7-2, lead screw; 7-3, guide rod; 8, housing; 9, camera bracket; 10, operating table; 11, microscope lens; 12, observation platform; 12-1, locking part; 12-2, clip; 12-3, elastic part; 12-4, light source; 12-5, platform body; 12-6, mounting hole; 12-7, rod; 12-8, limit part; 12-9, through hole; 13, base. DETAILED DESCRIPTION

[0038] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. 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 making creative efforts are within the scope of protection of the present invention.

[0039] It should be noted that in the description of the present invention, terms such as "upper", "lower", "left", "right", "inside", "outside", "front", "back", "center", "longitudinal", "lateral", "length", "width", "thickness", "vertical", "horizontal", "top", "bottom", "clockwise", and "counterclockwise" indicating directions or positional relationships are based on the directions or positional relationships shown in the accompanying drawings. This is merely for the convenience of description and does not indicate or imply that the device or element must have a specific orientation, be constructed and operate in a specific orientation. Therefore, it should not be understood as limiting the present invention. In addition, the terms "first", "second", "third", and "fourth" are used for descriptive purposes only and should not be understood as indicating or implying relative importance.

[0040] Furthermore, it should be noted that, in the description of the present invention, unless otherwise expressly specified or limited, the terms "disposed," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integral connections; mechanical connections or electrical connections; direct connections or indirect connections through an intermediate medium; and internal connections between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0041] The purpose of the present invention is to provide a micro-nano target in-situ detection device and method to solve the problems existing in the above-mentioned prior art, realize the in-situ direct optical image detection technology, and have resistance to environmental interference and high immediacy.

[0042] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0043] Example 1

[0044] like Figures 1 to 14 As shown, this embodiment provides a micro-nano target in-situ detection device 100, including a fluid delivery device 1, a microfluidic chip 3, an image acquisition device 4 and a detection device 5. The fluid delivery device 1 is connected to the microfluidic chip 3, and the fluid delivery device 1 can deliver the fluid at the target position 2 to the microfluidic chip 3; the image acquisition device 4 is used to connect the signal with the detection device 5, and the image acquisition device 4 is used to acquire the optical image of the micro-nano target in the fluid in the microfluidic chip 3; the detection device 5 can perform image recognition on the optical image. The present invention delivers the fluid at the target position 2 to the microfluidic chip 3 through the fluid delivery device 1, uses the microfluidic chip 3 as a sample carrier, and performs online sample processing through the microfluidic chip 3, thereby realizing in-situ sample collection and on-chip processing, and directly integrating the sample processing and preparation stage in traditional detection technology into the detection stage, thereby realizing its in-situ. At the same time, the microfluidic chip 3 has good light transmittance, so that the optical image of the micro-nano target can be obtained through the image acquisition device 4 and detected by the detection device 5, realizing the in-situ direct optical image detection technology, and having resistance to environmental interference and high immediacy.

[0045] In some specific embodiments, the detection device 5 is capable of identifying at least one micro-nano target, and the detection device 5 is capable of obtaining at least two types of feature information for each micro-nano target, the feature information including at least the category, content, contour shape, contour size, position coordinates, and area of ​​each micro-nano target. Faced with the need for simultaneous detection of multiple types of micro-nano targets in complex samples, such as the coexistence of white blood cells, red blood cells, and pathogens in blood, traditional detection technologies typically rely on manual labeling to distinguish different types of targets, which is inefficient. The detection device 5 of this embodiment is capable of identifying micro-nano targets in images, analyzing them, and outputting at least two types of feature information, such as the category, content, contour shape, contour size, position coordinates, and area of ​​the micro-nano targets, thereby achieving multi-target classification and recognition and multi-dimensional feature information output, completing in-situ detection, and enabling universal detection for multiple fields, improving detection efficiency, and providing effective support for research in multiple fields such as biomedical diagnosis and environmental pollutant monitoring.

[0046] In some specific embodiments, an observation platform 12 is also included, which includes a platform body 12-5, a clamp and a light source 12-4. A mounting hole 12-6 is provided on the working surface of the platform body 12-5, and the light source 12-4 is embedded in the mounting hole 12-6; the clamp is connected to the platform body 12-5, and the clamp can clamp the microfluidic chip 3 on the working surface; the light source 12-4 is arranged on the side of the microfluidic chip 3 away from the image acquisition device 4.

[0047] In some specific embodiments, the clamp includes two clips 12-2, two elastic components 12-3 and two locking components 12-1, and the clips 12-2, elastic components 12-3 and locking components 12-1 correspond to each other one by one; each elastic component 12-3 is arranged between the working surface and the corresponding clip 12-2; each clip 12-2 is fixedly connected to the observation plane to press the corresponding clip 12-2 and the corresponding elastic component 12-3 onto the working surface.

[0048] In some specific embodiments, each locking member 12-1 includes a rod 12-7 and a limit member 12-8, one end of each rod 12-7 is fixedly connected to the corresponding limit member 12-8, and the other end of each rod 12-7 is provided with a thread; a through hole 12-9 is provided on each clip 12-2; each rod 12-7 can pass through the through hole 12-9 of the corresponding clip 12-2 and the inner cavity of the corresponding elastic component 12-3 in sequence and be fixedly connected to the platform body 12-5. When the rod 12-7 is fixedly connected to the platform body 12-5, the limit member 12-8 can press the corresponding clip 12-2 and the corresponding elastic component 12-3 on the working surface, and the elastic component 12-3 is in a compressed state. Applying upward pressure to the end of the clip 12-2 away from the elastic member 12-3 can disengage the clip 12-2 from the work surface, allowing the microfluidic chip 3 to be removed or inserted. After the microfluidic chip 3 is placed on the work surface, the force applied to the clip 12-2 is released, allowing the clip 12-2 to contact the upper surface of the microfluidic chip 3 and clamp the microfluidic chip 3 to the work surface. This simple structure facilitates replacement of the microfluidic chip 3 and improves work efficiency.

[0049] In some embodiments, a lifting device 7 is further included. The image acquisition device 4 is disposed above the microfluidic chip 3. The observation platform 12 is connected to the lifting device 7. The lifting device 7 can move the observation platform 12 away from the image acquisition device 4 or toward the image acquisition device 4. By raising and lowering the observation platform 12 as needed, the distance between the observation platform 12 and the image acquisition device 4 can be adjusted to ensure image quality.

[0050] In some specific embodiments, a slider is further included. The lifting device 7 includes a drive device 7-1, a lead screw 7-2, and a guide rod 7-3. The observation platform 12 is fixedly connected to the slider; the slider is threadedly connected to the lead screw 7-2; and the slider is slidably connected to the guide rod 7-3. The drive device 7-1 can drive the lead screw 7-2 to rotate about the axis of the lead screw 7-2. By driving the lead screw 7-2 to rotate about the axis of the lead screw 7-2, the drive device 7-1 can cause the slider to move along the axial direction of the lead screw 7-2. The axes of the lead screw 7-2 and the guide rod 7-3 are parallel to each other.

[0051] In some specific embodiments, the image acquisition device 4 is a camera, and the lens of the image acquisition device 4 is a microscope lens 11, which has an image magnification function to capture optical images of micro-nano targets; the fluid delivery device 1 is a microfluidic pump. It should be noted that the image acquisition device 4 is not limited to achieving magnification through the microscope lens 11. A microscope can also be positioned between the image acquisition device 4 and the microfluidic chip 3 to achieve magnified photography of micro-nano targets. The magnification of the microscope lens 11 or microscope ranges from 1x to 100x.

[0052] In some specific embodiments, an electric field applying device is further included. The electric field applying device can apply an electric field to the flow channel of the microfluidic chip 3 to directionally align the micro-nano targets within the flow channel of the microfluidic chip 3. As a preferred embodiment, the electric field applying device includes two electrodes, one located on either side of the flow channel. A certain DC voltage is applied to the electrodes, causing the micro-nano targets to deflect along the direction of the electric field lines under the action of the electric field, thereby achieving directional alignment of the micro-nano targets.

[0053] In some specific embodiments, the microfluidic chip 3 has a pre-processing area 3-5 and a detection area 3-6. The pre-processing area 3-5 is used for on-chip processing of micro-nano targets, such as dispersing or directional arrangement of micro-nano targets to facilitate subsequent detection and analysis; the image acquisition device 4 is used to acquire optical images of the micro-nano targets in the detection area 3-6. The structural design of the pre-processing area 3-5 can be designed according to the control requirements of the micro-nano targets, or a physical field can be applied to the pre-processing area 3-5 according to the control requirements of the micro-nano targets. The microfluidic chip 3 has unique advantages such as a small amount of sample required, flexible flow field control, and smooth flow in the channel, which enables the micro-nano target in-situ detection device 100 to achieve in-situ sampling and on-chip sample processing, providing a basis for in-situ detection.

[0054] In some specific embodiments, the structural design of the pre-treatment areas 3-5 includes but is not limited to the use of serpentine channels to fully mix the micro-nano targets, or the use of spiral channels to orient the micro-nano targets; the external physical field includes but is not limited to the application of electric fields to enrich the micro-nano targets, or the application of ultrasonic fields to fully disperse the micro-nano targets.

[0055] In some embodiments, the microfluidic chip 3 has a three-layer structure, comprising a top plate 3-1 for packaging, a main plate 3-2 for providing a detection space, and a bottom plate 3-3 for packaging. The microfluidic chip 3 is made of a highly translucent polymer material such as PMMA, PVDF, PDMS, or PCB. The top plate 3-1 and bottom plate 3-3 are fixedly attached to the main plate 3-2 via thermocompression bonding, achieving a hermetic seal.

[0056] In some specific embodiments, the top plate 3-1, the main plate 3-2, and the bottom plate 3-3 are manufactured by laser cutting, mechanical engraving, etc., and the basic material is a square plate. For example, the basic material shape of the top plate 3-1, the main plate 3-2, and the bottom plate 3-3 is a 200mm*200mm square plate of different thicknesses; wherein through holes 12-9 are opened on both sides of the top plate 3-1 to form a sample buckle and a sample inlet 3-4, the main plate 3-2 is engraved by contour cutting to form a microchannel, and the bottom plate 3-3 is only cut for the outer shape without other processing, so as to maintain a high flatness and light transmittance of the bottom surface of the channel to improve image clarity.

[0057] In some specific embodiments, the top plate 3-1 is rectangular, and its dimensions are L2*W2*H3, H3=0.2mm–2mm; the main plate 3-2 is rectangular, and its dimensions are L2*W2*H4, H4=0.2mm–1mm; the microchannel width is W3, W3=0.05mm-1mm; the bottom plate 3-3 is rectangular, and its dimensions are L2*W2*H5, H5=1mm–6mm.

[0058] In some specific embodiments, the temperature of the hot pressing bonding is T, where T=80° C.-120° C.; the time of the hot pressing bonding is t, where t=30s-120s; and the number of hot pressing cycles is N, where N=2-10.

[0059] In some specific embodiments, the material of the shell 8 is a polymer material such as PMMA, PVDF, PCB, or a metal material such as aluminum, stainless steel, etc., the material of the base 13 is a metal material such as aluminum, stainless steel, etc., the thickness of the shell 8 is δ, δ = 3mm-10mm, and its internal space size is L*W*H.

[0060] In some embodiments, light source 12-4 has a convex structure for secure placement on platform body 12-5. The larger end of light source 12-4 measures D1*H1, while the smaller end measures D2*(H2-H1). Mounting hole 12-6 includes a recessed groove and a through hole 12-9 defined in platform body 12-5. The recessed groove connects to through hole 12-9, and the recessed groove has a larger diameter than through hole 12-9. The recessed groove is used to accommodate light source 12-4. The recessed groove measures D3*H1, where D3>D1. The through hole 12-9 has a diameter D4, where D1>D4>D2.

[0061] In some specific embodiments, a waste liquid collection tank 6 is further included. The waste liquid collection tank 6 is connected to the microfluidic chip 3 and is used to collect waste liquid flowing out of the microfluidic chip 3 .

[0062] In some embodiments, the detection device 5 includes a computer and an intelligent software system in the computer, which is used to detect micro-nano targets in situ. The intelligent software system detects micro-nano targets in the sample based on artificial intelligence technologies such as convolutional neural networks or Transformer neural networks.

[0063] In some specific embodiments, a camera bracket 9 is further included, which is arranged directly above the platform body 12 - 5 , and the image acquisition device 4 is fixedly connected to the camera bracket 9 .

[0064] In some specific embodiments, the device further includes a housing 8, a base 13, and an operating platform 10 for auxiliary operations. The lower end of the housing 8 is fixedly connected to the base 13, and the housing 8 and the base 13 form an installation space. The fluid delivery device 1, microfluidic chip 3, image acquisition device 4, detection device 5, lifting device 7, platform body 12-5, clamp, light source 12-4, camera bracket 9, operating platform 10, etc. are all arranged in the installation space. The lifting device 7 is arranged on one side of the housing 8, and the camera bracket 9 and platform body 12-5 are respectively arranged at the upper and lower ends of the same side of the lifting device 7. The operating platform 10 and the fluid delivery device 1 are arranged at the upper and lower ends of the other side of the housing 8. The operating platform 10 is fixedly connected to the side wall of the housing 8, and the fluid delivery device 1 is fixedly connected to the base 13 of the housing 8. The lifting device 7 and the camera bracket 9 are tightly connected to the housing 8. The operating platform 10 can be used to add additional power supplies and controllers, as well as adjust the internal hardware of the device.

[0065] In some specific embodiments, the elastic component 12 - 3 is a spring and the locking member 12 - 1 is a bolt.

[0066] Example 2

[0067] This embodiment provides a micro-nano target in-situ detection method based on the micro-nano target in-situ detection device 100 in Example 1, comprising the following steps:

[0068] S1. The fluid at the target position 2 is delivered to the microfluidic chip 3 by the fluid delivery device 1. The fluid delivery device 1 is a microfluidic pump. Driven by the microfluidic pump, a negative pressure is formed inside the microfluidic chip 3, which draws the sample at the target position 2, thereby achieving in-situ sampling of the micro-nano target and ensuring that the flow field inside the microfluidic chip 3 is relatively stable and uniform.

[0069] S2, processing the fluid entering the microfluidic chip 3 through the microfluidic chip 3;

[0070] S3, acquiring an optical image of the micro-nano target in the fluid processed by the microfluidic chip 3 through the image acquisition device 4;

[0071] S4. Perform image recognition on the optical image through the detection device 5.

[0072] In some embodiments, S1 further includes controlling the flow rate of the fluid delivery device 1 to be between 0.001 mL / min and 10 mL / min. Due to the small cross-sectional area of ​​the channels within the microfluidic chip 3, the microfluidic pump should be controlled to operate at a relatively low flow rate, typically between 0.001 mL / min and 10 mL / min, to avoid excessive flow rates within the channels that would prevent the capture of clear microscopic images.

[0073] In some specific embodiments, the sample input into the microfluidic chip 3 passes through the pre-processing area 3-5 to complete the on-chip processing of the micro-nano target; and then enters the detection area 3-6.

[0074] In some specific embodiments, S3 also includes: acquiring an optical image of the sample in the detection area 3-6 through the image acquisition device 4; the frequency of capturing the optical image by the image acquisition device 4 is controlled to be 1fps-1000fps to match the image processing speed of the intelligent software system.

[0075] In some specific embodiments, in order to obtain a sufficiently clear sample image while maintaining a moderate working focal length of the microscope lens 11 , the magnification of the microscope lens 11 should be controlled within a range of 1-100 times.

[0076] Example 3

[0077] This embodiment provides an in-situ detection method for micron-sized water droplets in a water-in-oil emulsion based on the micro-nano target in-situ detection device 100 of Example 1 and the micro-nano target in-situ detection method of Example 2. The main steps include:

[0078] The microfluidic chip 3 for inputting micro-nano targets is fixed on the platform body 12-5 embedded with the light source 12-4 by a clamp; the sample inlet and sample outlet 3-7 of the microfluidic chip 3 are connected to the target position 2 and the microfluidic pump respectively; the platform body 12-5 is fastened to the slider, and the image acquisition device 4 is controlled to move up and down by the driving device 7-1 (preferably a motor); the microscope lens 11 is installed on the camera; the oil-in-water emulsion in the sample bottle is sampled in situ using the microfluidic pump, and the low flow rate is controlled to be input into the microfluidic chip 3; the sample input into the microfluidic chip 3 passes through the pre-treatment area 3-5 to complete the on-chip processing of the sample; the fluid enters the chip detection area 3-6; the camera for capturing the optical image of the sample, which is fixed on the camera bracket 9 directly above the platform body 12-5, captures the microscopic image of the sample in the detection area 3-6 at a certain frequency, and transmits the image to the intelligent software system on the computer; the intelligent software system identifies the micro-nano targets in the sample according to the pre-trained model and the specified analysis mode, and analyzes and outputs the shape, position and content information.

[0079] The process conditions are:

[0080] The flow rate of the microfluidic pump is controlled at 0.4 mL / min; the magnification of the microscope lens 11 is 20 times; the image capture frequency of the camera is 2 fps; the size of the top plate 3-1 of the microfluidic chip 3 is 75 mm * 35 mm * 0.5 mm; the size of the main board 3-2 is 75 mm * 35 mm * 1 mm; the size of the bottom plate 3-3 is 75 mm * 35 mm * 2 mm, and the width of the microchannel is 1 mm; the size of the detection area 3-6 is 4 mm * 4 mm * 1 mm; the pre-treatment area 3-5 adopts a serpentine channel + spiral channel, so that the micro-nano targets are fully dispersed under the action of turbulence and then arranged in a directional manner, which is convenient for detection.

[0081] The experimental results are:

[0082] On the real-time monitoring interface of the intelligent software system, micron-sized water droplets are clearly visible; the average single image processing time of the intelligent software system is 43.2ms, the water droplet target recognition rate is 100%, and the sample water content detection accuracy is 92.7%.

[0083] Example 4

[0084] This embodiment provides an in-situ detection method for micron-sized water droplets in an oil-in-water emulsion based on the micro-nano target in-situ detection device 100 of Example 1 and the micro-nano target in-situ detection method of Example 2. The difference from Example 3 is that the flow rate of the microfluidic pump is replaced with 0.2 mL / min.

[0085] The experimental results are:

[0086] Compared with Example 3, the number of micron-sized water droplets appearing in the image increases, the average single image processing time is 103.6 ms, the water drop target recognition rate is 92.1%, and the sample water content detection accuracy is 88.7%.

[0087] Example 5

[0088] This embodiment provides an in-situ detection method for micron-sized water droplets in an oil-water emulsion based on the micro-nano target in-situ detection device 100 of embodiment 1 and the micro-nano target in-situ detection method of embodiment 2. The difference from embodiment 3 is that the flow rate of the microfluidic pump is replaced with 0.6 mL / min.

[0089] Experimental results:

[0090] Compared with Example 3, the number of micron-sized water droplets appearing in the image is reduced, the average single image processing time is 31.6 ms, the water drop target recognition rate is 100%, and the sample water content detection accuracy is 95.2%.

[0091] Example 6

[0092] This embodiment provides a method for detecting the behavior of micron-sized particles in particulate aerosols based on the micro-nano target in-situ detection device 100 of Example 1 and the micro-nano target in-situ detection method of Example 2. The difference from Example 3 is that: the sample bottle in the method is replaced with an aerosol sample enrichment pump, and the output information is changed to the particle chain length; an electric field application device (electrode) is added on both sides of the detection area 3-6 of the microfluidic chip 3 in the device, and an electric field with a voltage of 1500V is applied.

[0093] The experimental results are:

[0094] On the real-time monitoring interface of the intelligent software system, micron-sized particles are clearly visible, and their dynamic behaviors such as deposition, agglomeration and fracture under the action of the electric field can be fully observed; the average single image processing time of the intelligent software system is 16.3ms, the particle chain target recognition rate is 100%, and the particle chain length detection results are compared with the manually labeled calculation results, with a difference of 1.8%.

[0095] Example 7

[0096] This embodiment provides a method for detecting the behavior of micron-sized particles in particulate aerosols based on the micro-nano target in-situ detection device 100 of Example 1 and the micro-nano target in-situ detection method of Example 2. The difference from Example 6 is that the applied voltage is changed to 2000V.

[0097] The experimental results show that compared with Example 6, the particle chain length increases, and the intensity of behaviors such as deposition, agglomeration and breakage intensifies.

[0098] Example 8

[0099] This embodiment provides a method for detecting Escherichia coli based on the micro-nano target in-situ detection device 100 of Example 1 and the micro-nano target in-situ detection method of Example 2. The difference from Example 3 is that the oil-in-water emulsion is replaced by a fluid sample containing Escherichia coli; a fluorescent agent release section is added to the pretreatment area 3-5 of the microfluidic chip 3 in the device to fluorescently stain the Escherichia coli, and the light source 12-4 is replaced by a fluorescent light source 12-4.

[0100] The experimental results are: on the real-time monitoring interface of the intelligent software system, E. coli is clearly visible, and its dynamic behaviors such as peristalsis and migration can be fully observed.

[0101] The present invention uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only intended to help understand the method and core concept of the present invention. At the same time, those skilled in the art will find that the specific implementation methods and application scopes may vary based on the concept of the present invention. In summary, the contents of this specification should not be construed as limiting the present invention.

Claims

1. A micro-nano target in-situ detection device, characterized by: The system comprises a fluid delivery device, a microfluidic chip, an image acquisition device and a detection device. The fluid delivery device is connected to the microfluidic chip and can deliver fluid at a target position into the microfluidic chip. The image acquisition device is used for signal connection with the detection device and is used to acquire an optical image of a micro-nano target in the fluid in the microfluidic chip. The detection device can perform image recognition on the optical image.

2. The micro-nano target in-situ detection device according to claim 1, characterized in that: The detection device is capable of identifying at least one of the micro-nano targets, and the detection device is capable of obtaining at least two types of characteristic information of each of the micro-nano targets, wherein the characteristic information includes at least the category, content, outline shape, outline size, position coordinates and area of ​​each of the micro-nano targets.

3. The micro-nano target in-situ detection device according to claim 1, characterized in that: It also includes an observation platform, which includes a platform body, a clamp and a light source. A mounting hole is opened on the working surface of the platform body, and the light source is embedded in the mounting hole; the clamp is connected to the platform body, and the clamp can clamp the microfluidic chip on the working surface; the light source is arranged on the side of the microfluidic chip away from the image acquisition device.

4. The micro-nano target in-situ detection device according to claim 3, characterized in that: The clamp includes two clips, two elastic components and two locking components, and the clips, the elastic components and the locking components correspond to each other one by one; each elastic component is arranged between the working surface and the corresponding clip; each clip can press the corresponding clip and the corresponding elastic component onto the working surface by being fixedly connected to the observation plane.

5. The micro-nano target in-situ detection device according to claim 4, characterized in that: Each of the locking members includes a rod and a limiting member, one end of each of the rods is fixedly connected to the corresponding limiting member, and the other end of each of the rods is provided with a thread; a through hole is provided on each of the clips; each of the rods can pass through the through hole of the corresponding clip and the inner cavity of the corresponding elastic component in sequence and be fixedly connected to the platform body, and when the rod is fixedly connected to the platform body, the limiting member can press the corresponding clip and the corresponding elastic component tightly on the working surface.

6. The micro-nano target in-situ detection device according to claim 3, characterized in that: It also includes a lifting device, the image acquisition device is arranged above the microfluidic chip, the observation platform is connected to the lifting device, and the lifting device can drive the observation platform to move away from the image acquisition device or towards the image acquisition device.

7. The micro-nano target in-situ detection device according to claim 6, characterized in that: It also includes a slider, and the lifting device includes a driving device, a screw and a guide rod; the observation platform is fixedly connected to the slider; the slider is threadedly connected to the screw; the slider is slidingly connected to the guide rod; the driving device can drive the screw to rotate around the axis of the screw, and the driving device can make the slider move along the axial direction of the screw by driving the screw to rotate around the axis of the screw.

8. The micro-nano target in-situ detection device according to claim 1, characterized in that: The image acquisition device is a camera, and the lens of the image acquisition device is a microscope lens; the fluid delivery device is a microfluidic pump.

9. The micro-nano target in-situ detection device according to claim 1, characterized in that: The microfluidic chip further comprises an electric field applying device, which can apply an electric field to the flow channel of the microfluidic chip and directional arrange the micro-nano targets in the flow channel of the microfluidic chip.

10. A method for in-situ detection of micro-nano targets based on the in-situ detection device for micro-nano targets according to any one of claims 1 to 9, characterized in that: The steps include: S1, delivering the fluid at the target location to the microfluidic chip through the fluid delivery device; S2. Processing the fluid entering the microfluidic chip through the microfluidic chip; S3, acquiring, by the image acquisition device, the optical image of the micro-nano target in the fluid processed by the microfluidic chip; S4. Performing image recognition on the optical image by the detection device.

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