An automatic separation and purification method and system for two insects based on precise refractive index positioning

Abstract

The present invention relates to the technical field of water quality testing, and discloses a method for automatic separation and purification of two types of worms based on precise refractive index positioning, including: preparing and starting an automatic separation and purification device for two types of worms, which includes a liquid handling workstation. The liquid handling workstation includes centrifuge tubes, a liquid suction unit, a liquid transfer unit, and a controller. The controller controls the liquid suction unit and the liquid transfer unit to automatically complete precise liquid suction operations and precise liquid transfer operations of the water sample in the centrifuge tube based on an automated liquid handling control program; the liquid suction unit includes a liquid level sensor, a first flow sensor, and a first precision valve unit; the liquid transfer unit includes a second flow sensor and a second precision valve unit; collecting and obtaining a water sample and inputting it into the centrifuge tube of the automatic separation and purification device for two types of worms; precisely positioning and identifying the middle layer of the water sample based on the refractive index, and performing automatic separation and purification of the two types of worms based on the middle layer; after completing the automatic separation and purification of the two types of worms, cleaning all pipelines of the automatic separation and purification device for two types of worms. A corresponding system is also disclosed.

Description

Technical Field

[0001] The present invention relates to the technical field of water quality testing, and particularly to an automatic separation and purification method and system for Giardia and Cryptosporidium based on precise refractive index positioning. Background Art

[0002] Giardia lamblia and Cryptosporidium, collectively referred to as "two parasites", are water-borne, chlorine-resistant zoonotic pathogenic microorganisms that can penetrate conventional water treatment processes and enter the drinking water system. Human infection can lead to diarrhea, dehydration and even death. At present, there is no specific medicine for treatment, and it can only rely on the body's own immunity. To prevent large-scale infection events, it is particularly important to detect the two-parasite index in water. Now the membrane filtration concentration / density gradient separation fluorescence antibody method has been incorporated into GB / T 5750-2023 "Standard Test Methods for Drinking Water". The overall experimental process includes four steps: sample concentration, separation and purification, staining, and microscopic examination. At present, the products of the existing technologies have been able to achieve the intelligence and automation of the two stages of sample concentration and microscopic examination.

[0003] However, in the separation and purification stage, based on the principle of density gradient separation, in the past, whether adding a centrifugation medium or extracting the middle layer rich in two parasites, the professional and technical requirements for the operation of experimental personnel were very high. A slight mistake would lead to large experimental errors. In the lightest case, the recovery rate could not meet the standard, and in the worst case, impurities would be distributed in the middle layer, resulting in the inability to carry out the subsequent overall experiment. The relatively high practical technical threshold of this step has limited the overall development of this method in the industry to a certain extent. Summary of the Invention

[0004] The purpose of the present invention is to provide an automatic separation and purification method and system for two parasites based on precise refractive index positioning, which is used for the precise extraction of the middle layer by the two-parasite automatic separation and purification equipment, eliminating the microscopic examination experimental errors caused by the two-parasite automatic separation and purification, ensuring that the recovery rate meets the standard, and preventing the phenomenon of impurities being distributed in the middle layer.

[0005] The first aspect of the present invention lies in providing an automatic separation and purification method for two parasites based on precise refractive index positioning, including:

[0006] S1, prepare and start the two-parasite automatic separation and purification equipment. The two-parasite automatic separation and purification equipment includes a liquid processing workstation. The liquid processing workstation includes a centrifuge tube, a liquid suction unit, a liquid transfer unit and a controller. The controller has an automatic liquid processing control program inside. The controller is respectively connected to the liquid suction unit and the liquid transfer unit, and is used to control the liquid suction unit and the liquid transfer unit to automatically complete the precise liquid suction operation and precise liquid transfer operation of the water sample in the centrifuge tube based on the automatic liquid processing control program. The liquid suction unit includes a liquid level sensor, a first flow sensor and a first precision valve unit. The liquid transfer unit includes a second flow sensor and a second precision valve unit.

[0007] S2. Collect and obtain a water sample, and input the water sample into the centrifuge tube of the two-parasite automatic separation and purification device;

[0008] S3. Based on the refractive index, accurately locate and identify the middle layer of the water sample, and perform automatic separation and purification of the two parasites based on the middle layer;

[0009] S4. Based on the completion of the automatic separation and purification of the two parasites, clean all pipelines of the two-parasite automatic separation and purification device.

[0010] Preferably, the S3 includes:

[0011] S31. Measure the refractive indices of different layers in the centrifuge tube using a refractometer, and the different layers are determined according to a predetermined vertical spacing;

[0012] S32. Determine the refractive index corresponding to the middle layer based on the known density-refractive index relationship of different water samples, and accurately find the start and end positions of the middle layer based on the refractive index corresponding to the middle layer;

[0013] S33. Add a centrifugation medium, and perform automatic separation and purification of the two parasites based on the start and end positions of the middle layer. The automatic separation and purification of the two parasites includes precise liquid suction operation and precise pipetting operation on the water sample in the centrifuge tube.

[0014] Preferably, the range of the vertical spacing is 0.1 - 1 mm.

[0015] Preferably, the S31 includes:

[0016] (1) Align the light source of the refractometer with the centrifuge tube;

[0017] (2) Measure the refraction angles of the light rays formed by the light source when passing through different layers;

[0018] (3) Calculate the refractive indices of different layers in the centrifuge tube according to the refraction angles.

[0019] Preferably, the precise liquid suction operation includes:

[0020] (1) Precisely move the liquid suction unit to the position of the middle layer;

[0021] (2) Accurately suck the separation and purification liquid containing the two parasites in the middle layer;

[0022] Among them, the precise liquid suction operation is completed by a high-precision suction tool. The high-precision suction tool includes a suction head controlled by a stepping motor carried by itself. The precise movement of the liquid suction unit to the position of the middle layer is completed by the stepping motor carried by itself; the accurate suction of the separation and purification liquid containing the two parasites in the middle layer is completed by the suction head.

[0023] Preferably, the pipette tip is a horn-shaped pipette tip.

[0024] Preferably, the precise pipetting operation includes:

[0025] (1) Precisely move the pipetting unit to the outside of the centrifuge tube and above the container for receiving the separation and purification liquid containing the two worms;

[0026] (2) Release the separation and purification liquid containing the two worms into the container at a predetermined flow rate.

[0027] Preferably, the automatic liquid handling control program controls the liquid suction unit and the pipetting unit to automatically complete the precise liquid suction operation and the precise pipetting operation of the water sample in the centrifuge tube, including:

[0028] (1) Before liquid suction, the liquid level sensor monitors the liquid level height of the intermediate layer in real time and sends it to the controller. The controller controls the pipette tip to synchronously descend at a controlled speed based on the liquid level height of the intermediate layer, ensuring precise aspiration of a quantitative separation and purification liquid containing the two worms within a fixed time; wherein, controlling the pipette tip to synchronously descend at a controlled speed based on the liquid level height of the intermediate layer includes: obtaining the real-time measurement value of the liquid level sensor, and adjusting the descending speed of the pipette tip based on the PID control algorithm and the real-time measurement value of the liquid level sensor;

[0029] (2) During liquid suction, the controller automatically controls the pipette tip to aspirate the separation and purification liquid containing the two worms at a stable flow rate and speed based on the measurement value of the first flow sensor;

[0030] (3) During pipetting, the controller automatically controls the pipette tip to release the separation and purification liquid containing the two worms at a stable flow rate and speed based on the measurement value of the second flow sensor through the second precision valve unit.

[0031] Preferably, S4 includes:

[0032] S41, monitor the two-worm automatic separation and purification index in real time, and determine the completion of the two-worm automatic separation and purification based on the two-worm automatic separation and purification index reaching a predetermined threshold; the two-worm automatic separation and purification index is the two-worm recovery rate;

[0033] S42, based on the completion of the two-worm automatic separation and purification, start the automatic cleaning program and, according to the settings of the automatic cleaning program, automatically flush the pipeline, perform in-depth cleaning with water, buffer solution or cleaning solution, and dry.

[0034] The second aspect of the present invention is to provide a two-worm automatic separation and purification system based on precise refractive index positioning for implementing the method of the first aspect, including:

[0035] A two-worm automatic separation and purification equipment module (101), the two-worm automatic separation and purification equipment includes a liquid handling workstation, the liquid handling workstation includes centrifuge tubes, a liquid suction unit, a liquid transfer unit and a controller, the controller has an automated liquid handling control program inside, the controller is respectively connected to the liquid suction unit and the liquid transfer unit, and is used to control the liquid suction unit and the liquid transfer unit to automatically complete the precise liquid suction operation and precise liquid transfer operation of the water sample in the centrifuge tube based on the automated liquid handling control program; the liquid suction unit includes a liquid level sensor, a first flow sensor and a first precision valve unit; the liquid transfer unit includes a second flow sensor and a second precision valve unit;

[0036] A water sample collection and acquisition module (102), used to collect and acquire a water sample and input the water sample into the centrifuge tube of the two-worm automatic separation and purification equipment;

[0037] An intermediate layer positioning module (103), used to precisely locate and identify the intermediate layer of the water sample based on the refractive index, and perform two-worm automatic separation and purification based on the intermediate layer;

[0038] An automatic pipeline cleaning module (104), used to clean all pipelines of the two-worm automatic separation and purification equipment based on the completion of two-worm automatic separation and purification.

[0039] The third aspect of the present invention provides an electronic device, including a processor and a memory, the memory stores multiple instructions, and the processor is used to read the instructions and execute the method as described in the first aspect.

[0040] The fourth aspect of the present invention provides a computer-readable storage medium, the computer-readable storage medium stores multiple instructions, and the multiple instructions can be read and executed by the processor to execute the method as described in the first aspect.

[0041] The beneficial effects of the method, system, electronic device and computer-readable storage medium of the present invention:

[0042] (1) Greatly improve the detection efficiency, manifested in:

[0043] (1) Automated operation process: The equipment can automatically complete a series of operations for two-worm separation and purification, such as adding centrifugation medium and extracting the intermediate layer, etc., reducing the cumbersome process of manual operation, greatly saving operation time, and significantly improving the detection efficiency compared with traditional manual detection methods. For example, the work of adding separation medium that originally took twenty minutes or even longer can be completed within a few minutes using the automatic equipment.

[0044] (2) Batch processing capacity: Some two-worm automatic separation and purification equipment has the function of batch processing samples, and can process multiple samples simultaneously. For water companies, environmental monitoring agencies, etc. that need to conduct a large number of water sample detections, this can greatly improve work efficiency and meet the needs of large-scale detections.

[0045] (II) Enhanced detection accuracy:

[0046] (1) Precise control parameters: The automatic equipment adopts advanced sensors and control systems, and can precisely control various parameters in the separation and purification process, such as flow rate. These precise parameter controls can ensure that the separation and purification effect of the two worms is more stable and reliable, reduce errors caused by human factors, and improve the accuracy of detection results.

[0047] (2) Reduction of human contamination: During manual operation, samples may be contaminated due to non-standard operations of operators or environmental factors, etc., affecting the accuracy of detection results. The automatic separation and purification equipment operates in a closed system, which can effectively reduce the contamination of samples by the external environment and reduce the error of detection results.

[0048] (III) Reduction of labor intensity:

[0049] (1) Simplification of operation steps: Operators only need to perform simple operations, such as setting parameters, placing samples, etc., and the equipment can automatically complete the subsequent separation and purification work without complex manual operations, greatly reducing the labor intensity of operators. Especially for those who need to conduct two-worm detection work for a long time, using automatic equipment can effectively relieve work fatigue.

[0050] (2) No need for professional skills: Compared with traditional two-worm separation and purification methods that require operators to have high professional skills and rich experience, the operation of automatic separation and purification equipment is relatively simple and can be mastered after simple training, reducing the professional requirements for operators.

[0051] (IV) Having stable operating performance, practical, safe and reliable: The automatic separation and purification equipment has undergone strict testing and optimization, has stable operating performance, and can maintain a good working state during long-term operation to ensure the smooth progress of detection work.

[0052] In summary, due to many technical advantages such as high automation level, high detection accuracy, and strong processing ability, the two-worm automatic separation and purification equipment has important application value in application fields such as disease control and prevention centers, water supply systems, third-party testing, environmental monitoring, and water quality testing institutions. Moreover, the popularization and application of this product provide a basic tool for a large number of drinking water detections; it provides support for the prevention and control of "two worms" in the water supply safety guarantee from source to tap, and provides guarantee for curbing the spread route of "two worms" in tap water, with high social value. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the related art, the following will briefly introduce the drawings required to be used in the description of the specific embodiments or the related art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0054] Figure 1 It is a flowchart of the two-worm automatic separation and purification method based on precise positioning by refractive index according to an embodiment of the present invention;

[0055] Figure 2 It is a flowchart of step S3 of the two-worm automatic separation and purification method based on precise positioning by refractive index according to an embodiment of the present invention;

[0056] Figure 3 It is a flowchart of step S4 of the two-worm automatic separation and purification method based on precise positioning by refractive index according to an embodiment of the present invention;

[0057] Figure 4 It is a schematic diagram of the principle architecture of the two-worm automatic separation and purification system based on precise positioning by refractive index according to an embodiment of the present invention;

[0058] Figure 5 It is a structural diagram of an electronic device according to an embodiment of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0059] The following will clearly and completely describe the technical solutions of the present invention with reference to the drawings. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art without creative efforts based on the embodiments of the present invention belong to the scope of protection of the present invention.

[0060] In the description of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings. It is 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 should not be construed as a limitation to the present invention. In addition, the terms "first", "second", "third" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance.

[0061] In the description of the present invention, it should be noted that unless otherwise clearly specified and limited, the terms "installed", "connected", "connected" 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 directly connected or indirectly connected through an intermediate medium, and it can be the communication inside 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 situations. Embodiment 1

[0062] See Figure 1 , this embodiment provides a two-worm automatic separation and purification method based on precise refractive index positioning, including:

[0063] S1. Prepare and start the two-worm automatic separation and purification equipment. The two-worm automatic separation and purification equipment includes a liquid processing workstation. The liquid processing workstation includes a centrifuge tube, a liquid suction unit, a liquid transfer unit, and a controller. The controller has an automated liquid processing control program inside. The controller is respectively connected to the liquid suction unit and the liquid transfer unit, and is used to control the liquid suction unit and the liquid transfer unit to automatically complete the precise liquid suction operation and precise liquid transfer operation of the water sample in the centrifuge tube based on the automated liquid processing control program; the liquid suction unit includes a liquid level sensor, a first flow sensor, and a first precision valve unit; the liquid transfer unit includes a second flow sensor and a second precision valve unit;

[0064] S2. Collect and obtain a water sample, and input the water sample into the centrifuge tube of the two-worm automatic separation and purification equipment;

[0065] S3. Based on precise refractive index positioning, identify the middle layer of the water sample, and perform two-worm automatic separation and purification based on the middle layer;

[0066] As Figure 2 shown, as a preferred embodiment, the S3 includes:

[0067] S31. Measure the refractive indices of different layers in the centrifuge tube using a refractometer. The different layers are determined according to a predetermined vertical spacing. In this embodiment, the range of the vertical spacing is 0.1 - 1 mm;

[0068] As a preferred embodiment, S31 includes:

[0069] (1) Align the light source of the refractometer with the centrifuge tube;

[0070] (2) Measure the refraction angles of the light rays formed by the light source when passing through different layers;

[0071] (3) Calculate the refractive indices of different layers in the centrifuge tube based on the refraction angles.

[0072] In this embodiment, in density gradient centrifugation, layers with different densities may have different refractive indices. Therefore, the layer interfaces can be identified by the changes in refractive indices, and then the position of the intermediate layer can be determined. A refractometer is used to measure the refractive indices of different layers in the centrifuge tube. Align the light source of the refractometer with the centrifuge tube and measure the refraction angles of the light rays when passing through different layers. The refractometer calculates the refractive indices based on the refraction angles.

[0073] Principle of operation: Refractive index is the ratio of the speed of light in a vacuum to the speed of light in the medium. The higher the refractive index of a material, the stronger the ability to refract incident light. The higher the refractive index, the thinner the lens, that is, for lenses with the same center thickness, the same degree, and the same material, the lens with a higher refractive index is thinner at the edge than the one with a lower refractive index. The refractive index is closely related to the electromagnetic properties of the medium. When light travels from a relatively optically denser medium to a relatively optically less dense medium and the incident angle is greater than the critical angle, total internal reflection can occur. In density gradient centrifugation, the essential reason why layers with different densities may have different refractive indices is that the refractive index is also closely related to the arrangement of ions (including the physicochemical manifestations of the solution containing the two organisms). For isotropic optical materials, such as amorphous (non-crystalline) and cubic crystals, there is only one refractive index; when light enters an anisotropic medium, it generally divides into two waves with mutually perpendicular vibration directions and unequal propagation speeds, and they have two refracted light rays respectively, constituting so-called double refraction. For these two refracted light rays, the refractive index of the light ray parallel to the incident plane is called the ordinary refractive index; regardless of how the incident angle of the incident light changes, it is always a constant and obeys the law of refraction. The refractive index formed by the other light ray perpendicular to the incident plane changes with the direction of the incident light and is called the extraordinary refractive index; it does not obey the law of refraction. When light is incident along the crystal optic axis direction, only the ordinary refractive index exists. When incident perpendicular to the optic axis direction, the extraordinary refractive index reaches its maximum value, which is a characteristic of the material. Thus, the extraordinary refractive index is larger along the direction with a greater degree of crystal close packing.

[0074] S32. Determine the refractive index corresponding to the intermediate layer based on the known density-refractive index relationship of different water samples, and accurately find the start and end positions of the intermediate layer based on the refractive index corresponding to the intermediate layer;

[0075] S33. Add a centrifugation medium and perform automatic separation and purification of the two worms based on the start and end positions of the intermediate layer. The automatic separation and purification of the two worms includes precise liquid suction operations and precise pipetting operations performed on the water sample in the centrifuge tube;

[0076] As a preferred embodiment, the precise liquid suction operation includes:

[0077] (1) Precisely move the liquid suction unit to the position of the intermediate layer;

[0078] (2) Accurately suck the separation and purification liquid containing the two worms in the intermediate layer.

[0079] As a preferred embodiment, the precise liquid suction operation is completed by a high-precision suction tool. The high-precision suction tool includes a suction head controlled by a stepping motor carried by itself. The precise movement of the liquid suction unit to the position of the intermediate layer is completed by the stepping motor carried by itself, and the movement accuracy can reach the millimeter level or even higher; the accurate sucking of the separation and purification liquid containing the two worms in the intermediate layer is completed by the suction head; wherein, the shape of the suction head is designed to be helpful for accurate extraction. For example, in this embodiment, a horn-shaped suction head is used, which fits better with the intermediate layer during the suction process and reduces the interference with the substances in the upper and lower layers.

[0080] As a preferred embodiment, the precise pipetting operation includes:

[0081] (1) Precisely move the pipetting unit to the outside of the centrifuge tube and above the container for receiving the separation and purification liquid containing the two worms;

[0082] (2) Release the separation and purification liquid containing the two worms into the container at a predetermined flow rate.

[0083] As a preferred embodiment, the automatic liquid handling control program controls the liquid suction unit and the pipetting unit to automatically complete the precise liquid suction operation and the precise pipetting operation of the water sample in the centrifuge tube, including:

[0084] (1)Before liquid suction, the liquid level height of the intermediate layer is monitored in real time by a liquid level sensor and sent to the controller, and the controller controls the suction head to synchronously descend at a controlled speed based on the liquid level height of the intermediate layer, ensuring that a quantitative separation and purification liquid containing two insects is accurately aspirated within a fixed time; wherein, controlling the suction head to synchronously descend at a controlled speed based on the liquid level height of the intermediate layer includes: obtaining the real-time measurement value of the liquid level sensor, and adjusting the descending speed of the suction head based on the PID control algorithm and the real-time measurement value of the liquid level sensor, avoiding inhaling substances from other layers, and realizing efficient and accurate liquid transfer;

[0085] (2)During liquid suction, the controller automatically controls the suction head to aspirate the separation and purification liquid containing two insects at a stable flow rate and speed based on the measurement value of the first flow sensor through the first precision valve unit, thereby avoiding mixing substances from other layers due to too fast suction speed;

[0086] (3)During liquid transfer, the controller automatically controls the suction head to release the separation and purification liquid containing two insects at a stable flow rate and speed based on the measurement value of the second flow sensor through the second precision valve unit.

[0087] S4. Based on the completion of the automatic separation and purification of two insects, all pipelines of the two-insect automatic separation and purification equipment are cleaned.

[0088] As Figure 3 shown, as a preferred embodiment, the S4 includes:

[0089] S41. Monitor the two-insect automatic separation and purification index in real time, and determine the completion of the automatic separation and purification of two insects based on the two-insect automatic separation and purification index reaching a predetermined threshold;

[0090] In this embodiment, the two-insect automatic separation and purification index is the two-insect recovery rate.

[0091] S42. Based on the completion of the automatic separation and purification of two insects, start the automatic cleaning program and, according to the settings of the automatic cleaning program, automatically flush the pipelines, perform in-depth cleaning with water, buffer solution or cleaning solution, and dry.

[0092] In this embodiment, the method and system have an automatic cleaning system, which can thoroughly clean and dry the pipelines according to the program settings, effectively preventing cross-contamination. The system supports a comprehensive cleaning process, including automatic flushing, drying, in-depth cleaning with water or a special buffer solution (cleaning solution). In addition, the system also provides functions such as delayed start, pause, and resume, ensuring the flexibility and convenience of operation. The complete collection function enables the samples after cleaning to be safely and accurately collected for subsequent use. Embodiment 2

[0093] As Figure 4 shown, this embodiment provides a two-worm automatic separation and purification system based on precise refractive index positioning for implementing the method of Embodiment 1, including:

[0094] A two-worm automatic separation and purification equipment module 101. The two-worm automatic separation and purification equipment includes a liquid processing workstation. The liquid processing workstation includes a centrifuge tube, a liquid suction unit, a liquid transfer unit, and a controller. The controller has an automated liquid processing control program inside. The controller is respectively connected to the liquid suction unit and the liquid transfer unit for controlling the liquid suction unit and the liquid transfer unit to automatically complete the precise liquid suction operation and precise liquid transfer operation of the water sample in the centrifuge tube based on the automated liquid processing control program. The liquid suction unit includes a liquid level sensor, a first flow sensor, and a first precision valve unit. The liquid transfer unit includes a second flow sensor and a second precision valve unit;

[0095] A water sample collection and acquisition module 102 for collecting and acquiring a water sample and inputting the water sample into the centrifuge tube of the two-worm automatic separation and purification equipment;

[0096] An intermediate layer positioning module 103 for precisely positioning and identifying the intermediate layer of the water sample based on the refractive index and performing two-worm automatic separation and purification based on the intermediate layer;

[0097] An automatic pipeline cleaning module 104 for cleaning all pipelines of the two-worm automatic separation and purification equipment based on the completion of two-worm automatic separation and purification.

[0098] The present invention also provides a memory storing multiple instructions for implementing the method as in Embodiment 1.

[0099] As Figure 5 shown, the present invention also provides an electronic device including a processor 301 and a memory 302 connected to the processor 301. The memory 302 stores multiple instructions that can be loaded and executed by the processor so that the processor can execute the method as in Embodiment 1.

[0100] Through the description of the above embodiments, those skilled in the art can clearly understand that the above embodiments can be implemented by software or by means of software plus a necessary general hardware platform. Based on such an understanding, the technical solutions of the above embodiments can be embodied in the form of a software product. The software product can be stored in a non-volatile storage medium (which can be a CD-ROM, a USB flash drive, a mobile hard disk, etc.) and includes several instructions for causing a computer device (which can be a personal computer, a server, or a network device, etc.) to execute the methods described in various embodiments of the present invention.

[0101] 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 them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for automatic separation and purification of two insects based on precise positioning of refractive index, characterized in that: include: S1, prepare and start the automatic separation and purification equipment of two insects, the automatic separation and purification equipment of two insects includes a liquid processing workstation, the liquid processing workstation includes a centrifuge tube, a liquid suction unit, a liquid transfer unit and a controller, the controller has an automated liquid processing control program inside, the controller is connected with the liquid suction unit and the liquid transfer unit respectively, and is used to control the liquid suction unit and the liquid transfer unit to automatically complete the precise liquid suction operation and precise liquid transfer operation of the water sample in the centrifuge tube based on the automated liquid processing control program; the liquid suction unit includes a liquid level sensor, a first flow sensor and a first precision valve unit; the liquid transfer unit includes a second flow sensor and a second precision valve unit; S2, collecting and obtaining a water sample, and inputting the water sample into the centrifuge tube of the two-worm automatic separation and purification device; S3, accurately locating and identifying the middle layer of the water sample based on the refractive index, and automatically separating and purifying the two insects based on the middle layer; S4, based on the completion of the automatic separation and purification of the two insects, cleaning all pipelines of the automatic separation and purification equipment of the two insects; The S3 includes: S31, measuring the refractive index of different layers in the centrifuge tube using a refractometer, wherein the different layers are determined according to a predetermined vertical spacing; S32, determining the refractive index corresponding to the middle layer based on the known density-refractive index relationship of different water samples, and accurately finding the starting and ending positions of the middle layer based on the refractive index corresponding to the middle layer; S33, adding centrifugal medium, and automatically separating and purifying the two insects based on the starting and ending positions of the intermediate layer, wherein the automatic separation and purification of the two insects includes precise aspiration and precise pipetting operations on the water sample in the centrifuge tube.

2. The method for automatic separation and purification of two insects based on precise refractive index positioning according to claim 1 is characterized in that: The vertical spacing ranges from 0.1 to 1 mm.

3. The method for automatic separation and purification of two insects based on precise refractive index positioning according to claim 2 is characterized in that: The S31 includes: (1) Aligning the light source of the refractometer toward the centrifuge tube; (2) measuring the refraction angle of the light generated by the light source when passing through different layers; (3) Calculating the refractive index of different layers in the centrifuge tube according to the refraction angle.

4. The method for automatic separation and purification of two insects based on precise refractive index positioning according to claim 3 is characterized in that: The precise liquid aspiration operation includes: (1) Move the liquid suction unit precisely to the position of the middle layer; (2) accurately aspirating the separation and purification solution containing the two insects in the middle layer; Among them, the precise liquid suction operation is completed by a high-precision suction tool, and the high-precision suction tool includes a suction head controlled by a stepper motor carried by itself, and the stepper motor carried by itself is used to accurately move the liquid suction unit to the position of the middle layer; the suction head is used to accurately suck the separation and purification liquid containing the two insects in the middle layer.

5. The method for automatic separation and purification of two insects based on precise refractive index positioning according to claim 4 is characterized in that: The suction head is a trumpet-shaped suction head.

6. The method for automatic separation and purification of two insects based on precise refractive index positioning according to claim 5 is characterized in that: The precise pipetting operation includes: (1) accurately moving the pipetting unit to the outside of the centrifuge tube and above the container receiving the separation and purification solution containing the two insects; (2) releasing the separated and purified liquid containing the two insects into the container at a predetermined flow rate.

7. The method for automatic separation and purification of two insects based on precise refractive index positioning according to claim 6 is characterized in that: The method of controlling the aspiration unit and the pipetting unit based on the automated liquid handling control program to automatically complete the precise aspiration operation and precise pipetting operation of the water sample in the centrifuge tube includes: (1) Before aspirating the liquid, the liquid level of the intermediate layer is monitored in real time by a liquid level sensor and sent to a controller, and the controller controls the suction head to descend synchronously at a controlled speed based on the liquid level of the intermediate layer, so as to ensure accurate aspiration of a quantitative amount of the separated and purified liquid containing the two insects within a fixed time; wherein the controlling the suction head to descend synchronously at a controlled speed based on the liquid level of the intermediate layer comprises: obtaining a real-time measurement value of the liquid level sensor, and adjusting the descending speed of the suction head based on a PID control algorithm and the real-time measurement value of the liquid level sensor; (2) During the liquid aspiration process, the controller automatically controls the suction head to aspirate the separation and purification liquid containing the two insects at a stable flow rate and speed through the first precision valve unit based on the measurement value of the first flow sensor; (3) During the pipetting process, the controller automatically controls the pipette tip to release the separation and purification solution containing the two worms at a stable flow rate and speed through the second precision valve unit based on the measurement value of the second flow sensor.

8. The method for automatic separation and purification of two insects based on precise refractive index positioning according to claim 7 is characterized in that: The S4 includes: S41, real-time monitoring of the automatic separation and purification index of the two insects, and determining that the automatic separation and purification of the two insects is completed based on the automatic separation and purification index of the two insects reaching a predetermined threshold; the automatic separation and purification index of the two insects is the recovery rate of the two insects; S42, based on the completion of the automatic separation and purification of the two insects, start the automatic cleaning program and according to the settings of the automatic cleaning program, automatically flush the pipeline, use water, buffer solution or cleaning solution for deep cleaning and drying.

9. An automatic separation and purification system for two insects based on precise positioning of refractive index, used to implement the method described in any one of claims 1 to 8, characterized in that: include: An automatic separation and purification equipment module (101) for two insects, the automatic separation and purification equipment for two insects comprising a liquid processing workstation, the liquid processing workstation comprising a centrifuge tube, a liquid aspiration unit, a liquid transfer unit and a controller, the controller having an automated liquid processing control program inside, the controller being connected to the liquid aspiration unit and the liquid transfer unit respectively, and being used to control the liquid aspiration unit and the liquid transfer unit to automatically complete precise liquid aspiration and precise liquid transfer operations of the water sample in the centrifuge tube based on the automated liquid processing control program; the liquid aspiration unit comprising a liquid level sensor, a first flow sensor and a first precision valve unit; the liquid transfer unit comprising a second flow sensor and a second precision valve unit; A water sample collection and acquisition module (102) is used to collect and acquire water samples and input the water samples into the centrifuge tube of the two-worm automatic separation and purification device; An intermediate layer positioning module (103) is used to accurately locate and identify the intermediate layer of the water sample based on the refractive index, and automatically separate and purify the two insects based on the intermediate layer; The automatic pipeline cleaning module (104) is used to clean all pipelines of the automatic separation and purification device for the two insects based on the completion of the automatic separation and purification of the two insects.

Images