CN116338156A - Liquid path system for analyzing microparticles by using multi-methodology - Google Patents

Abstract

The invention relates to the field of detection and analysis of microparticles, in particular to a liquid path system for analyzing microparticles by using multiple methodologies, which comprises a sampling module, a cleaning sleeve module, a reaction cup module, a planar sheath flow module, a fluorescent sheath flow module and a cleaning module, wherein the sampling module samples microparticles by adopting a sampling needle to obtain a sample; the cleaning sleeve module cleans the sampled sampling needle; the reaction cup module dilutes and fluorescent dyes the sample; the planar sheath flow module forms a planar sheath flow from the fluorescent dyed sample; the fluorescent sheath flow module forms a particle sheath flow; the cleaning module cleans the liquid path, so that the planar sheath flow required by the image method detection technology and the particle sheath flow required by the fluorescent flow type cell detection technology can be completed only by one sample collection, the detection advantages of the two methodologies can be well combined, and the detection advantages of the two methodologies can be realized by less sampling amount, lower instrument cost and reagent cost _、 And more comprehensive information of various microparticles in the urine sample can be obtained in a faster time.

Description

Liquid path system for analyzing microparticles by using multi-methodology

Technical Field

The invention relates to the field of detection and analysis of microparticles, in particular to a liquid path system for analyzing microparticles by using multiple methodologies.

Background

The information of the number and the morphology of the micro-particles (including red blood cells, white blood cells, epithelial cells, crystals, tubes, bacteria and the like) in the urine sample has important significance for clinical diagnosis, and the detection and analysis of the micro-particles in the urine sample are one of the most common and necessary examination items in clinical work.

At present, the detection and analysis technologies of microparticles in urine samples are mainly divided into two main categories: image detection technology and fluorescence flow cytometry.

The image method detection technology is developed on the basis of microscopic examination, and can be specifically classified into precipitation detection and planar laminar flow detection according to a liquid flow control mode of a sample and a detection container. The precipitation detection principle is basically similar to the detection principle of an artificial microscope, and microparticles (formed components) in a urine sample are rapidly precipitated in a container with fixed thickness through control of a liquid path system, and then a microscopic amplification shooting system is matched for shooting a picture of the precipitated microparticles. The plane laminar flow detection principle is to control the liquid path by using a plane laminar flow technology, so that a certain thickness of laminar flow is formed in a specific container by a urine sample, particles in the urine rapidly pass through the laminar flow, and then a high-frequency flash lamp and a microscopic amplifying shooting system are matched to capture a picture of the particles. The two detection methods can obtain microparticle pictures which are similar to microscopic examination, then a series of processing is carried out on the shot microparticle pictures by a computer, and the computer carries out intelligent classification (comprising red blood cells, white blood cells, epithelial cells, crystals, tubes, bacteria and the like) on the microparticles according to the size, the shape and other characteristics of various microparticles, so that the detection result of the microparticles in urine is obtained.

The image method detection technology has the main advantages that the detection principle is similar to that of the traditional microscopic examination (the current gold standard for detecting the micro-particles in urine), the micro-particle picture similar to that of the microscopic examination can be obtained, and the detection result can be checked conveniently by a checking doctor. Along with the development of artificial intelligence technology, the image method detection technology is rapidly developed, and the number and accuracy of intelligent identification and classification of the microparticles are obviously improved. According to the nominal parameters of related instruments, the intelligent identification and classification of the microparticles in urine can reach up to 30 types at present, and great reference value is provided for clinical diagnosis. The main disadvantage of the image detection technology is that the detection speed is relatively slow, in order to ensure the detection speed of the instrument, the volume of the urine sample to be detected is limited, and the actual detection volume of the sample of the mainstream product is basically less than 2 mu L, so that the detection performance of rare particles (such as various tube types) in the urine sample cannot be ensured. In addition, since the angles, focal lengths, and the like of the microparticles in the imaging sample do not completely coincide with microscopic examination, the microparticles cannot be accurately and accurately classified from the obtained images of the microparticles.

Fluorescent flow cytometry has been developed. The detection technology is that after micro particles in urine are dyed by fluorescent pigment, single and longitudinal cell flows are formed under the action of sheath fluid; when the microparticles pass through the laser detection area, the instrument can detect the change of scattered light and fluorescence; after the instrument detects signals such as forward scattered light intensity, forward scattered light pulse width, side scattered light intensity, fluorescence pulse width and the like, the instrument comprehensively recognizes and calculates the size, length, intracellular complexity, cytoplasmic dyeability, cytoplasmic length and the like of the microparticles through a series of processes; and then the microparticles are intelligently classified (comprising red blood cells, white blood cells, epithelial cells, crystals, tubes, bacteria and the like), and corresponding scatter diagrams are made to obtain quantitative detection results of the microparticles.

The fluorescent flow type cell detection technology has the main advantages of high detection speed, larger actual detection volume of urine samples, and the actual detection volume of samples of the main flow products is basically larger than 5 mu L, which is more than 2 times of the actual detection volume of the image detection technology, and can ensure the detection performance of small numbers of microparticles (such as various tube types) in the urine samples. In addition, the flow control mode of the flow cytometry is to form the microparticles in the urine into a single cell flow, so that all the microparticles passing through the detection area can be detected, and the flow control mode has great advantages for detecting the high-concentration urine sample. The main defects of the nucleic acid fluorescence staining flow cytometry are that the microparticle classification types are less, the nominal microparticle classification types of the related instruments are less than 15 items at present, and the method has a larger gap with the nominal 30 items of the related instruments of the imaging method detection technology; furthermore, the nucleic acid fluorescence staining flow cytometry cannot obtain a picture of the particles, and a checking doctor cannot directly check the detection result; in addition, the crystallization in urine has larger interference on laser signals, and the accuracy of the detection result of the urine sample with a large amount of crystallization is relatively low. The invention can complete the planar sheath flow required by the image method detection technology and the particle sheath flow required by the fluorescent flow type cell detection technology through one sample collection, thereby improving the detection simplicity.

Disclosure of Invention

The invention aims to provide a liquid path system for carrying out microparticle analysis by applying multiple methodologies, which aims to carry out microparticle analysis faster and improve detection quality.

In order to achieve the above object, in a first aspect, the present invention provides a liquid path system for performing microparticle analysis by applying multiple methodologies, including a sampling module, a cleaning sleeve module, a reaction cup module, a planar sheath flow module, a fluorescent sheath flow module and a cleaning module, wherein the sampling module is connected with the planar sheath flow module, the fluorescent sheath flow module is connected with the planar sheath flow module, the reaction cup module is connected with the fluorescent sheath flow module, the cleaning sleeve module is connected with the reaction cup module, and the cleaning module is connected with the planar sheath flow module;

the sampling module is used for sampling the microparticles by adopting a sampling needle to obtain a sample;

the cleaning sleeve module is used for cleaning the sampled sampling needle;

the reaction cup module is used for diluting and fluorescent staining a sample;

the plane sheath flow module is used for forming a plane sheath flow from the sample after fluorescent staining;

the fluorescent sheath flow module is used for forming a particle sheath flow;

the cleaning module is used for cleaning the liquid path.

The sampling module comprises a sampling needle, a mixing unit, a filtering unit, a liquid level detection unit and a conductivity detection unit, wherein the sampling needle is used for sampling, and the mixing unit repeatedly sucks and spits samples in a test tube through a syringe so as to achieve mixing; the liquid level detection unit is used for judging whether the sampling needle drops down to the liquid level of the sample; the conductivity detection unit detects conductivity of the sample.

Wherein, the sampling needle has detachable filter screen, the filter screen is used for preventing impurity entering system.

The cleaning sleeve module comprises a cleaning unit and an air blowing unit, wherein the cleaning unit is used for cleaning the inner wall and the outer wall of the sampling needle, and the air blowing unit is used for removing hanging liquid on the outer wall of the sampling needle.

The reaction cup module comprises a dilution unit and a dyeing unit, wherein the dilution unit is used for diluting a sample, and the dyeing unit is used for dyeing the sample.

The reaction cup module further comprises a mixing unit and a heating unit, wherein the mixing unit is used for uniformly mixing the sample by adopting a stirring rod, and the heating unit is used for heating and preserving heat of the sample.

The planar sheath flow module comprises a planar sheath flow device and a first injector, wherein the planar sheath flow device is controlled by pressure, so that a sample forms a stable laminar flow under the pressure of sheath flow, and the thickness and the position of the laminar flow of the sample can be controlled by adjusting the pressure of sheath liquid and the speed of pushing the sample by the injector.

The fluorescent sheath flow module comprises a particle sheath flow device and a second injector, the sample is stably injected into the sheath flow by the second injector, so that the sample forms stable particle flow under the pressure of the sheath flow, and the pressure of the sheath liquid and the speed of pushing the sample by the sample injector are regulated.

The invention discloses a liquid path system for analyzing microparticles by applying multiple methodologies, which comprises a sampling module, a cleaning sleeve module, a reaction cup module, a planar sheath flow module, a fluorescent sheath flow module and a cleaning module, wherein the sampling module is connected with the planar sheath flow module, the fluorescent sheath flow module is connected with the planar sheath flow module, the reaction cup module is connected with the fluorescent sheath flow module, the cleaning sleeve module is connected with the reaction cup module, and the cleaning module is connected with the planar sheath flow module; the sampling module is used for sampling the microparticles by adopting a sampling needle to obtain a sample; the cleaning sleeve module is used for cleaning the sampled sampling needle; the reaction cup module is used for diluting and fluorescent staining a sample; the plane sheath flow module is used for forming a plane sheath flow from the sample after fluorescent staining; the fluorescent sheath flow module is used for forming a particle sheath flow; the cleaning module is used for cleaning the liquid path. The liquid path system of the invention analyzes the micro-particles, only needs one sample collection to finish the planar sheath flow required by the image method detection technology and the particle sheath flow required by the fluorescent flow type cell detection technology, can well integrate the detection advantages of two methodologies, obtains more comprehensive information of various micro-particles in the urine sample with less sampling amount, lower instrument cost and reagent cost and faster time, and provides enough reference information for clinical diagnosis.

Drawings

In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

Fig. 1 is a block diagram of a fluid path system for performing microparticle analysis using a multi-methodology according to a first embodiment of the present invention.

Fig. 2 is a block diagram of a liquid path system for analyzing microparticles using a multi-methodology according to a second embodiment of the present invention.

A 1-sampling module, a 2-cleaning sleeve module, a 3-reaction cup module, a 4-plane sheath flow module, a 5-fluorescence sheath flow module and a 6-cleaning module.

Detailed Description

Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative and intended to explain the present invention and should not be construed as limiting the invention.

First embodiment

Referring to fig. 1, fig. 1 is a schematic diagram of a liquid path system for performing microparticle analysis using multi-methodology according to a first embodiment of the present invention. The invention provides a liquid path system for analyzing microparticles by applying multiple methodologies, which comprises:

the device comprises a sampling module 1, a cleaning sleeve module 2, a reaction cup module 3, a planar sheath flow module 4, a fluorescent sheath flow module 5 and a cleaning module 6, wherein the sampling module 1 is connected with the planar sheath flow module, the fluorescent sheath flow module 5 is connected with the planar sheath flow module 4, the reaction cup module 3 is connected with the fluorescent sheath flow module 5, the cleaning sleeve module 2 is connected with the reaction cup module 3, and the cleaning module 6 is connected with the planar sheath flow module 4;

the sampling module 1 is used for sampling microparticles by adopting a sampling needle to obtain a sample;

the cleaning sleeve module 2 is used for cleaning the sampled sampling needle;

the reaction cup module 3 is used for diluting and fluorescent staining a sample;

the plane sheath flow module 4 is used for forming a plane sheath flow from the sample after fluorescent staining;

the fluorescent sheath flow module 5 is used for forming a particle sheath flow;

the cleaning module 6 is used for cleaning the liquid path.

In this embodiment, the liquid path system mainly comprises a sampling module 1, a cleaning sleeve module 2, a reaction cup module 3, a planar sheath flow module 4, a fluorescent sheath flow module 5 and a cleaning module 6, and is mainly characterized by comprising a planar sheath flow device (an imaging detection channel can be realized by using a sedimentation detection technology or a planar laminar flow detection technology, wherein the detection speed of the sedimentation detection technology is relatively low, the component cost is relatively low, the detection speed of the planar laminar flow detection technology is relatively high, the detection speed can be matched with the detection speed of a nucleic acid fluorescent staining flow type cell detection channel, the component cost is relatively high, and an instrument can select one of the detection technologies as an imaging detection channel according to different design requirements.

The sampling module 1 comprises a sampling needle, a mixing unit, a filtering unit, a liquid level detection unit and a conductivity detection unit, wherein the sampling needle is used for sampling, and the mixing unit repeatedly sucks and spits samples in a test tube through a syringe so as to achieve mixing; the liquid level detection unit is used for judging whether the sampling needle drops down to the liquid level of the sample; the conductivity detection unit detects conductivity of the sample.

The sampling needle has a removable filter screen for preventing contaminants from entering the system.

In this embodiment, the sampling needle module mainly includes a mixing system, a filtering structure, a liquid level detection sensing module, a sample presence detection module, and a conductivity detection module. The mixing system repeatedly sucks and spits samples in the test tube through the injector so as to achieve the mixing effect; the filter screen which can be replaced is designed in the sampling needle structure, so that larger particles and other impurities in a sample are prevented from entering a liquid path system, the phenomenon that an instrument is failed due to blockage of a sheath flow device and other liquid path components is avoided, a function of backflushing and cleaning the filter screen is designed in a liquid path, the probability of filter screen blockage is reduced, the filter screen is designed into a replaceable structure, and the filter screen can be taken out for replacement or cleaning under the condition of blockage; the liquid level detection sensing function can judge whether the sampling needle drops down to the liquid level of the sample; the liquid level detection function can be combined with the needle setting distance of the sampling needle to judge the sample height in the test tube, so as to calculate whether the sample exists in the test tube or not and whether the sample quantity meets the minimum sample quantity requirement of the current sampling; the conductivity detection structure can detect the conductivity of the sample, and can further judge whether the sampling amount is enough for the test, so that the information of whether the sample exists or not is prompted.

Further, the cleaning sleeve module 2 comprises a cleaning unit and an air blowing unit, wherein the cleaning unit is used for cleaning the inner wall and the outer wall of the sampling needle, and the air blowing unit is used for cleaning hanging liquid on the outer wall of the sampling needle.

In this embodiment, the cleaning sleeve module 2 is mainly used for cleaning the inner and outer walls of the sampling needle, and meanwhile, an air blowing function is designed in the cleaning sleeve structure, so that hanging liquid on the outer wall of the sampling needle can be cleaned, the accuracy of sample sucking and spitting of the sampling needle is ensured, and the carrying pollution rate of the test sample is ensured to meet the use requirement.

The cuvette module 3 comprises a dilution unit for diluting the sample and a staining unit for staining the sample.

The reaction cup module 3 further comprises a mixing unit and a heating unit, wherein the mixing unit is used for uniformly mixing the sample by adopting a stirring rod, and the heating unit is used for heating and preserving heat of the sample.

In this embodiment, the reaction cup module 3 is mainly used for diluting and fluorescent staining a sample, and is designed with a stirring rod for uniformly mixing and heating and heat-preserving functions, so that the sample can repeatedly react with fluorescent dye, and the accuracy and repeatability of the test are ensured. After the sample is injected into the reaction cup module 3 through the sampling needle, the sample is diluted by injecting diluent, and meanwhile, the stirring and mixing function is started, so that the sample is diluted and some impurities (such as crystallization, amorphous phosphate, mucus wires and the like) possibly affecting the test in the sample are dissolved; injecting dye liquor after diluting the sample, and dyeing a specific example in the sample; the reaction cup module 3 is designed with heating and heat preservation functions, and can heat the diluent and the whole reaction cup so as to ensure sample dilution, uniform mixing and stability of dye liquor, and further ensure stability of test results.

Further, the planar sheath flow module 4 comprises a planar sheath flow device and a first injector, the planar sheath flow device is controlled by pressure, so that a stable laminar flow is formed by the sample under the pressure of the sheath flow, and the thickness and the position of the laminar flow of the sample can be controlled by adjusting the pressure of the sheath liquid and the speed of pushing the sample by the injector.

In this embodiment, the planar sheath flow module 4 mainly includes a planar sheath flow device, a syringe, and other auxiliary components to form a stable planar sheath flow. The sheath fluid used for forming the sheath flow is controlled by pressure (can also be realized by other structures such as an injector, a peristaltic pump and the like), the sample is stably injected into the sheath flow by the injector, so that the sample forms a stable laminar flow under the pressure of the sheath flow, and the thickness and the position of the laminar flow of the sample can be controlled by adjusting the pressure of the sheath fluid and the speed of pushing the sample by the sample injector. Microparticles in urine pass through the laminar flow quickly, then a high-frequency flash lamp and a microscopic amplifying shooting system (generally formed by combining an optical microscope and a digital camera) are matched to capture microparticle pictures, a series of treatments are carried out on the shot microparticle pictures through a computer to obtain the sizes, the forms and other characteristics of various microparticles, finally the microparticles are intelligently classified by using an artificial intelligence technology, and meanwhile, microparticle pictures similar to microscopic examination can be provided, so that a checking doctor can check the detection result conveniently.

Further, the fluorescence sheath flow module 5 comprises a particle sheath flow device and a second injector, the sample is smoothly injected into the sheath flow by the second injector, so that the sample forms stable particle flow under the pressure of the sheath flow, and the pressure of the sheath liquid and the speed of pushing the sample by the sample injector are regulated.

In this embodiment, the fluorescent sheath flow module 5 mainly comprises a particle sheath flow device, a syringe and other auxiliary components to form a stable particle sheath flow. The pressure of the sheath fluid used for forming the sheath flow is controlled (the sheath fluid can also be realized by other structures such as a syringe, a peristaltic pump and the like), the sample is stably injected into the sheath flow by the syringe, so that the sample forms stable particle flow under the pressure of the sheath flow, and the speed of the particle flow in the sheath flow can be regulated by regulating the pressure of the sheath fluid and the speed of the sample syringe pushing the sample. Microparticles in the sample form single and column cell streams under the action of sheath fluid; when the microparticles pass through the laser detection area, the instrument can detect the change of scattered light and fluorescence; after the instrument detects signals such as forward scattered light intensity, forward scattered light pulse width, side scattered light intensity, fluorescence pulse width and the like, the instrument comprehensively recognizes and calculates to obtain the information such as the size, the length, the complexity of the inside of cells, the dyeability of cytoplasm and the length of cytoplasm of the microparticles through a series of processing, and makes a corresponding scatter diagram. According to different requirements of sample analysis, cost control requirements of instruments and the like, different colorants and different lasers can be selected to form a detection channel, and scattered light detection devices at other angles and fluorescence detection devices at different wavelengths can be added to obtain more information of the microparticles.

The cleaning module 6 mainly comprises a waste liquid pump, a cleaning pressure system and the like, and is used for cleaning liquid path components and pipelines of a sampling needle, a reaction cup, a planar sheath flow device and a particle sheath flow device through which a sample passes, so that cross contamination is avoided.

The liquid path system of the invention is used for analyzing the microparticles, the planar sheath flow required by the image method detection technology and the particle sheath flow required by the fluorescent flow type cell detection technology can be completed only by one sample acquisition, the detection advantages of the two methodologies can be well integrated, the more comprehensive information of various microparticles in the urine sample can be obtained with less sampling amount, lower instrument cost and reagent cost and faster time, and enough reference information can be provided for clinical diagnosis.

The liquid path system of the invention is mainly used for analyzing the microparticles, so that the actual detection volume of a sample can be increased under the condition of not reducing the detection speed, the advantages of an image detection technology and a fluorescence flow cytometry detection technology can be integrated, the more comprehensive information of various microparticles in the sample can be obtained, and enough reference information can be provided for clinical diagnosis.

As described above, the image method detection technology can obtain the microparticle image which is detected by the similar microscope, so that the detection result can be checked by the checking doctor conveniently; the artificial intelligence technology can be utilized to intelligently identify and classify the micro particles in the sample to 30 kinds or more; however, in order to ensure the test speed of the instrument, the actual test volume of the sample is limited, and the actual test volume of the sample of the mainstream product is basically smaller than 2 mu L, so that the detection performance of rare particles (such as various tube types) in the urine sample cannot be ensured; in addition, since the angles, focal lengths, etc. of the microparticles in the sample photographed by the imaging method are not completely consistent with the microscopic examination, the microparticles cannot be accurately and completely classified according to the obtained picture of the microparticles; most importantly, some microparticles are difficult to distinguish from pictures and morphology, such as erythrocytes and single shed yeasts.

As described above, the fluorescence flow cytometry is to form the microparticles in the sample into a single cell flow, so that all the microparticles passing through the detection area can be detected, and the fluorescence flow cytometry has great advantages for detecting high-concentration urine samples; the detection speed is faster, the volume of the detection sample is more, and the actual detection volume of the sample of the mainstream product is basically more than 5 mu L at present; more importantly, the fluorescent staining flow cell detection channel can stain cell membranes and cell nuclei to obtain more comprehensive information of the microparticles, so that the problem of intelligent classification errors caused by incomplete consistency of angles, focal lengths and the like of the microparticles with microscopic examination in a sample picture shot by an imaging method is solved; the detection channel sensitivity can be adjusted to detect the microparticle information with different sizes, so that microparticles such as epithelial cells and tubular cells with the size of more than 10 microns can be detected, and bacteria with the size of about 2 microns can be detected, however, the interference of crystals in urine on laser signals is large, and the accuracy of the detection result of a urine sample with a large number of crystals is relatively low.

Specifically, some micro-particles are difficult to distinguish from images and forms by using an image detection technology, for example, erythrocytes and single exfoliated yeasts, but the yeasts with nucleic acid can be dyed by using a fluorescent dye cell detection technology, so that the yeasts with nucleic acid can be well identified and classified.

Specifically, in the fluorescent staining cell detection technology, the interference of crystals in the urine sample on the laser signal is large, and the accuracy of the detection result of the urine sample with a large number of crystals is relatively low; however, by using the image method detection technology, not only the effect of crystallization can be avoided, but also the picture of crystallization can be shot, and the crystallization can be intelligently identified and classified, and more than 10 kinds of crystallization can be automatically identified by the existing artificial intelligence technology.

Specifically, the concentration of bacteria in urine samples with urinary tract infection is high, and the bacteria are difficult to segment and identify by using an image detection technology; however, by using the fluorescent staining cell detection technology, nucleic acid in bacteria can be stained and detected independently, so that more information of the bacteria can be obtained, and the clinical examination and judgment can be facilitated.

Specifically, aiming at the detection of the white blood cells, the existing image detection technology cannot sub-classify the white blood cells, but the white blood cells can be classified into lymphocytes, monocytes, neutrophils, basophils, eosinophils and the like by the fluorescent staining cell detection technology, so that more comprehensive information is provided for clinical diagnosis.

Specifically, aiming at the detection of red blood cells, the current fluorescent staining cell detection technology cannot sub-classify the red blood cells, but can divide the red blood cells into normal red blood cells, shadow red blood cells, echinocyte, small red blood cells and the like by the image detection technology, thereby providing more comprehensive information for clinical diagnosis.

In summary, the liquid path system of the invention is applied to analyze the micro particles, so that the actual detection volume of the sample can be improved under the condition of not reducing the detection speed, and according to the current technical level, under the condition of 100 detection speeds per hour, the actual detection volume of the sample of the liquid path system of the invention is about 7 mu L, which is obviously improved compared with 2 mu L (image detection technology) and 5 mu L (fluorescence flow detection technology) of the existing unilateral law, the detection performance of micro particles (such as various tube types) with fewer numbers in urine samples can be ensured, and the accuracy and repeatability of the test result can be well ensured; more mainly, the liquid path system is used for analyzing the microparticles, the image detection technology of the liquid path system can be used for obtaining microparticle pictures similar to microscopic examination, the artificial intelligent technology is used for ensuring the automatic identification and classification quantity of the microparticles, a doctor can conveniently check the detection result, meanwhile, the size, the length, the volume, the chromatin length and other information of the microparticles can be obtained through the fluorescent staining cell detection technology, a corresponding scatter diagram is made, finally, the detection results of the two detection technologies are integrated to obtain more comprehensive information of the microparticles, and the microparticles are intelligently classified (including red blood cells, white blood cells, epithelial cells, crystals, tubular bacteria and the like), so that more comprehensive and valuable information is provided for clinical diagnosis.

When the liquid path system is used for analyzing the microparticles, the sampling amount is less than that of the method for independently detecting the microparticles, and the detection result of the double methods can be obtained only by sample acquisition of the single methodology (the single sampling amount of the current main stream product is about 1 mL); the liquid path system can detect the two forensics, but the whole machine frame and components can be reused, such as a sampling mechanism, a mixing mechanism, a cleaning system and the like, the instrument cost is not greatly increased compared with that of the instrument of the single forensics, but the testing of the two forensics can be completed, and the cost performance of the instrument is high; and compared with the single fluorescence flow cytometry technology, the reagent required by the instrument is not increased, and the use amount of the reagent is not obviously increased because of multiplexing and optimization of a liquid path, and the use cost of a user is not obviously increased.

Second embodiment

Referring to fig. 2, the following fig. 2 is a specific embodiment of the present invention, which is merely illustrative of the principles, and some functions may be added or reduced according to actual needs, and some functions may achieve equivalent or similar effects by other means.

Firstly, judging whether the sample is enough to finish the test or not through a liquid level detection function. Specifically, the sampling needle moves to the position right above the test tube port, then the needle is put down, the liquid level detection function is started at this moment, whether the sampling needle touches the sample liquid level is detected, when the sampling needle touches the sample liquid level, the control system judges whether the sample in the test tube is enough to finish the test through the needle put down height of the sampling needle, if not, the test is fed back to the software end, the user is prompted that the sample amount is less, and the sampling work cannot be finished.

And when the sample amount in the test tube is judged to be enough to finish the test, uniformly mixing the samples in the test tube. Specifically, when the sample amount in the test tube is judged to be enough to finish the test, the sampling needle moves downwards to the bottom of the test tube, and the injector M1 is communicated with the sampling needle to repeatedly suck and spit the sample for a plurality of times, so that the particles precipitated in the sample are fully and uniformly mixed.

After the samples were thoroughly mixed, sampling was started. Specifically, the sample enters the pipeline through the filter and the V1 valve under the action of the injector M1 until the sample quickly fills the pipeline between the V4 valve and the V5 valve; the sample between the V4 valve and the V5 valve is used for detecting a planar sheath flow channel; the sample between the V4 valve and the V3 valve is used for detecting the conductivity; the sample between the V4 valve and the V3 valve is separated into samples to the reaction cup through the V2 valve and the sampling needle under the action of the injector M2, and is used for detecting a fluorescence flow type channel; all samples to be detected pass through the filter, so that the blocking probability of the liquid path system is reduced.

After the sampling is completed, the detection of the planar sheath flow channel can be started. Specifically, a fixed amount of sheath fluid is poured into the planar sheath fluid tank before the test, and a stable positive pressure is established, and the pressure value can be detected and controlled by the pressure sensor PS 2; under the action of positive pressure PS2, sheath liquid in the planar sheath flow tank enters the planar sheath flow device through the V24 valve, and stable sheath flow is formed in the planar sheath flow device; meanwhile, the samples in the V5-V4 sections also enter the planar sheath flow device under the action of the injector M1, and stable laminar flow is formed under the wrapping of sheath fluid. Microparticles in a sample pass through the laminar flow rapidly, then a high-frequency flash lamp and a microscopic amplifying shooting system (generally formed by combining an optical microscope and a digital camera) are matched to capture microparticle pictures, a series of processing is carried out on the shot microparticle pictures through a computer to obtain the sizes, the forms and other characteristics of various microparticles, finally an artificial intelligence technology is used for intelligently classifying the microparticles, and meanwhile, microparticle pictures similar to microscopic examination can be provided, so that a checking doctor can check the detection result conveniently.

After sampling is finished, the sampling needle moves into the reaction cup while the planar sheath flow channel is detected, samples in the V1-V3 sections are separated from V2 into the reaction cup through the sampling needle by the injector M2, and a certain amount of diluent is required to be discharged into the reaction cup in advance; after sample separation, adding diluent, mixing uniformly, fully mixing and reacting the sample with the diluent, and dissolving particles which do not need to be tested or are possibly infected with the test in the sample, such as amorphous phosphate, crystallization or mucus wire; after fully mixing, standing for about 5 seconds, adding a fixed amount (15 mu L) of dye liquor by a quantitative pump M5, and continuously mixing uniformly to fully mix and react the sample in the reaction cup with the dye liquor. In the whole diluting and dyeing process, the reaction cup device is heated and kept at a fixed temperature (40 degrees) in practice, so that the test result is not influenced by the external environment change, and the accuracy and the repeatability of the test result are ensured.

After the sample is diluted and stained in the cuvette, detection of the fluorescence flow channel can be started. Specifically, a certain amount of sheath liquid is poured into the sheath liquid heating device in advance, the sheath liquid is heated to a fixed temperature (35 ℃), stable positive pressure is established, and the pressure value can be detected and controlled by the pressure sensor PS 1; the positive pressure in the positive pressure tank enables sheath liquid pre-stored in the sheath liquid heating device to enter the particle sheath flow device through the V19 valve, meanwhile, a sample in the reaction cup is pushed to a sample inlet of the particle sheath flow device through the injector M2, and at the moment, the sample is pushed to the particle sheath flow device through the injector M3, so that single and longitudinal cell flows are formed under the action of the sheath liquid; when the microparticles pass through the laser detection area, the instrument can detect the change of scattered light and fluorescence; after the instrument detects signals such as forward scattered light intensity, forward scattered light pulse width, side scattered light intensity, fluorescence pulse width and the like, the instrument comprehensively recognizes and calculates to obtain the information such as the size, the length, the complexity of the inside of cells, the dyeability of cytoplasm and the length of cytoplasm of the microparticles through a series of processing, and makes a corresponding scatter diagram. According to different requirements of sample analysis, cost control requirements of instruments and the like, different colorants and different lasers can be selected to form a detection channel, and scattered light detection devices at other angles and fluorescence detection devices at different wavelengths can be added to obtain more information of the microparticles.

After the test is completed, the cleaning system starts to clean the pipelines and the liquid path components of each section. In this case, the sheath liquid is directly used for cleaning, and a cleaning liquid can be additionally added. The sheath liquid for cleaning is poured into the sheath liquid tank through the sheath liquid pump, and then the sheath liquid in the sheath liquid tank is pushed to each section of pipeline under the action of the pressure of the sheath liquid tank, so that the effect of cleaning the pipeline is achieved. The filter and the reaction cup with higher blockage probability are designed with the function of backflushing and cleaning, so that the blockage probability of a liquid path system is reduced.

The above disclosure is only a preferred embodiment of the present invention, and it should be understood that the scope of the invention is not limited thereto, and those skilled in the art will appreciate that all or part of the procedures described above can be performed according to the equivalent changes of the claims, and still fall within the scope of the present invention.

Claims (8)

1. A liquid path system for analyzing microparticles by using multiple methodologies is characterized in that,

the device comprises a sampling module, a cleaning sleeve module, a reaction cup module, a planar sheath flow module, a fluorescent sheath flow module and a cleaning module, wherein the sampling module is connected with the planar sheath flow module, the fluorescent sheath flow module is connected with the planar sheath flow module, the reaction cup module is connected with the fluorescent sheath flow module, the cleaning sleeve module is connected with the reaction cup module, and the cleaning module is connected with the planar sheath flow module;

the sampling module is used for sampling the microparticles by adopting a sampling needle to obtain a sample;

the cleaning sleeve module is used for cleaning the sampled sampling needle;

the reaction cup module is used for diluting and fluorescent staining a sample;

the plane sheath flow module is used for forming a plane sheath flow from the sample after fluorescent staining;

the fluorescent sheath flow module is used for forming a particle sheath flow;

the cleaning module is used for cleaning the liquid path.

2. A fluid path system for microparticle analysis using a multi-methodology according to claim 1,

the sampling module comprises a sampling needle, a mixing unit, a filtering unit, a liquid level detection unit and a conductivity detection unit, wherein the sampling needle is used for sampling, and the mixing unit repeatedly sucks and spits samples in a test tube through an injector so as to achieve mixing; the liquid level detection unit is used for judging whether the sampling needle drops down to the liquid level of the sample; the conductivity detection unit detects conductivity of the sample.

3. A fluid path system for microparticle analysis using a multi-methodology according to claim 2,

the sampling needle has a removable filter screen for preventing contaminants from entering the system.

4. A fluid path system for microparticle analysis using a multi-methodology according to claim 3,

the cleaning sleeve module comprises a cleaning unit and an air blowing unit, wherein the cleaning unit is used for cleaning the inner wall and the outer wall of the sampling needle, and the air blowing unit is used for cleaning hanging liquid on the outer wall of the sampling needle.

5. A fluid path system for performing microparticle analysis using a multi-methodology according to claim 4,

the reaction cup module comprises a dilution unit and a dyeing unit, wherein the dilution unit is used for diluting a sample, and the dyeing unit is used for dyeing the sample.

6. A fluid path system for performing microparticle analysis using a multi-methodology according to claim 5,

the reaction cup module further comprises a mixing unit and a heating unit, wherein the mixing unit is used for uniformly mixing the sample by adopting a stirring rod, and the heating unit is used for heating and preserving heat of the sample.

7. A fluid path system for performing microparticle analysis using a multi-methodology according to claim 6,

the planar sheath flow module comprises a planar sheath flow device and a first injector, wherein the planar sheath flow device is controlled by pressure, so that a sample forms a stable laminar flow under the pressure of sheath flow, and the thickness and the position of the laminar flow of the sample can be controlled by adjusting the pressure of sheath liquid and the speed of pushing the sample by the injector.

8. A fluid path system for microparticle analysis using a multi-methodology according to claim 7,

the fluorescent sheath flow module comprises a particle sheath flow device and a second injector, the sample is stably injected into the sheath flow by the second injector, so that the sample forms stable particle flow under the pressure of the sheath flow, and the pressure of the sheath liquid and the speed of pushing the sample by the sample injector are regulated.

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