Micro-fluidic chip for detecting urinary sediment, intelligent dyeing detection device and intelligent dyeing method

Abstract

The invention relates to a micro-fluidic chip for detecting urinary sediment, an intelligent dyeing detection device and an intelligent dyeing method. The device has the advantages that the fluid mechanics principle of urine particles is utilized, the micro-column array is designed, high-adhesion treatment is combined, capillary channel movement is utilized, the fluid movement direction of urine sediment passing through the micro-column arrays with different sizes and densities is controlled on the microscale, urine sediment components with different particle sizes and different forms are separated, and the urine sediment separation effect is achieved. Non-invasive intelligent separation of particles in urine liquid and enrichment of visible components in different chip positions are realized; the enrichment and concentration of urine cells of an ultra-large urine sample volume (greater than 100ml) are automatically realized, the detection rate of microorganisms such as tumor cells and mycobacterium tuberculosis is greatly improved, and the situation that a small sample detection result cannot represent the real urinary sediment characteristics of a patient is avoided; the urine sediment components are automatically separated without centrifugation, so that the completeness of urine cells is ensured, and the damage of urine centrifugation to the cells and other visible components is prevented.

Description

Technical Field

[0001] The present invention relates to the technical field of urinary sediment detection, and in particular to a microfluidic chip for detecting urinary sediment, an intelligent staining detection device and an intelligent staining method. Background Art

[0002] Kidney resident cells are mainly composed of vascular endothelial cells, epithelial cells and renal interstitial cells. Among them, epithelial cells include glomerular epithelial cells, renal capsule epithelial cells and renal tubular epithelial cells.

[0003] The glomerular epithelium outside the glomerular basement membrane, namely podocytes, is an important part of the glomerular filtration barrier. Under physiological conditions, podocytes not only participate in the selective filtration of proteins by the filtration barrier, but also participate in maintaining the normal function and structure of glomerular capillary loops. Under pathological conditions, podocyte damage can not only directly lead to the occurrence of proteinuria, but also be closely related to the formation of glomerulosclerosis. Recent studies have confirmed that podocyte injury is the initiating link in the formation of glomerulosclerosis. Since podocytes are terminally differentiated cells and do not have the characteristics of proliferation and differentiation, their exfoliation and damage are irreversible. Therefore, the exfoliation and damage of glomerular podocytes mean glomerulosclerosis.

[0004] Renal tubular epithelial injury is often the main site of damage by various pathogenic factors, and exfoliated renal tubular epithelium can often be detected in urine. Under physiological conditions, renal tubular epithelium mainly reabsorbs water, electrolytes, glucose and amino acids, maintains acid-base electrolyte balance, produces erythropoietin and active vitamin D, etc. Pathological damage often causes its exfoliation, but renal tubular epithelial hyperplasia and repair are relatively fast.

[0005] Renal capsule epithelial cells are often involved in the formation of crescents. Therefore, detecting the excretion characteristics of cells from different sources in urine can not only determine the site of injury, but also be used to judge the degree of injury and prognosis.

[0006] Importance of urinary sediment detection: Urinary sediment is the basic content of urine analysis, and its diagnostic value is as follows:

[0007] (1) Examination of the number and atypia of red blood cells in urine is the key basis for determining nephritis;

[0008] (2) Leukocyturia is an important indicator for the diagnosis of urinary tract infections;

[0009] (3) An increase in urothelial cells helps to judge the site of injury;

[0010] (4) The presence of pathological casts in urine is the basis for diagnosing the existence of kidney diseases;

[0011] (5) Determine urinary tract tumors;

[0012] (6)An increase in eosinophils in urine suggests drug-induced interstitial nephritis, etc.;

[0013] (7)The detection of molds in fresh urine often indicates dysbacteriosis or contamination of the body;

[0014] (8)The detection of various crystals is helpful for the diagnosis of urolithiasis, drug metabolism disorders, metabolic nephropathy, etc.

[0015] Therefore, this timely, non-invasive, and simple method of urine cell excretion analysis is of great significance for understanding the laws of kidney diseases and kidney injuries, and urine sediment examination has important clinical diagnostic value.

[0016] Urine analysis includes two methods: urine flow cytometry automated analysis and microscopic examination. In 1983, IRIS Corporation in the United States developed and produced the world's first "Yollow IRIS" automatic urine sediment analyzer. In 1995, Sysmex Corporation in Japan combined flow cytometry, analog image, and impedance technology [see Figure 1 , and developed a new generation of UF-100 fully automatic urine sediment analyzer. In 1997, Dysis Corporation in the United States launched the flow cell type R / S2003 urine sediment analysis workstation, opening the precedent of digital image analysis of urine sediment; in 2002, IRIS Corporation in the United States launched the IQ-200 fully automatic urine sediment analyzer with a urine sediment digital imaging system. In 2007, Sysmex Corporation in Japan launched the UF-1000i fully automatic urine analyzer using a red semiconductor laser. Recently, there have emerged urine automated analyzers such as CombiScan500 from Germany, Elektronika 77 (Langmai Urised) from Hungary, and AX-4030 from ARKRAY in Japan. The research and development of urine automated analyzers in China started in the early 2000s. After 2002, imitation instruments from companies such as Chongqing Tianhai (US-2026) and Changsha Aivai (AVE-76) were successively launched, and only the IQ-200 from IRIS Corporation in the United States has obtained the certification of the US FDA.

[0017] However, at present, there are no dedicated automated analysis instruments suitable for the precise qualitative and quantitative analysis of urine cells at home and abroad. Manual microscopy is still the "gold standard" for urine sediment examination. Urine cell analysis at home and abroad has remained at the level of non-staining qualitative detection, with high false positives or false negatives, and it is impossible to perform precise qualitative and quantitative analysis.

[0018] Since abnormal results of urine cell analysis often lead to misdiagnosis by clinicians, in the future, urine sediment analysis will enter a new era that combines general staining techniques for urine cell morphology, urine cell molecular recognition techniques, and automated staining and quantitative techniques, thus breaking through the strange circle of relying solely on single traditional microscopic morphology staining or single fluorescence staining full-automatic analysis of flow cytometry for urine cell analysis, and greatly improving the clinical diagnostic accuracy and efficiency of urine sediment automated analysis.

[0019] The main disadvantages of existing urine autoanalyzers include:

[0020] 1) Defects in the recognition principle design: The UF-1000i stains urinary sediment particles with a nuclear fluorescent dye and performs computer simulation recognition based on the physical characteristics of the fluorescence intensity, impedance, and scattered light of urinary sediment particles; the IQ-200 takes a phase-contrast image of the local black-and-white urinary particles flowing under a phase-contrast microscope and magnifies them to 400 times for computer simulation recognition. Neither of them has a complete physical image of urinary sediment;

[0021] 2) Without color staining of urinary sediment, resulting in the loss of information such as hyaline casts in urinary sediment;

[0022] 3) The test results are easily interfered with by the interaction between the components of urinary sediment;

[0023] 4) Without the recognition of marker molecules of urinary sediment particles: Epithelial cells, tumor cells, etc. that are diagnostically significant cannot be effectively recognized;

[0024] 5) The urine sample is not concentrated, and 100 - 200 μl of urine is not evenly sampled, making it difficult to represent the complete information of the patient's urinary sediment;

[0025] 6) The urinary sediment autoanalyzer has a high recognition sensitivity (>90%) and a specificity lower than 20%;

[0026] 7) Urinary cells are affected by other components of urine. Only general staining is used for cell morphology recognition. Since there is no specific identification of urinary cell types, the origin of diabetic renal epithelial cells cannot be determined, so qualitative and quantitative diagnostic research of clinical significance cannot be carried out by general microscopic examination;

[0027] 8) Without the concept of stratified analysis of urinary cells: The components of urinary sediment are complex. After pretreatment of urine analysis and stratified extraction, the interaction of urinary cell analysis will be greatly reduced, which is a prerequisite for subsequent automated urinary sediment analysis.

[0028] 9) Without the technology of thin-layer preparation of exfoliated cells in highly viscous urine. Thin-layer cell preparation has been achieved in vaginal exfoliated cell analysis. However, due to the damage of the cell membrane of urinary cells, their adhesion is greatly reduced, and the shedding rate during spreading and staining is high, making automatic analysis impossible to complete;

[0029] 10) Without a precise quantitative method for urinary cell classification: Currently, the international urinary sediment analysis is mainly based on two types of urinary sediment quantitative analysis methods developed based on different urinary sediment counting and staining methods.

[0030] The main advantage of manual microscopic examination is accurate qualitative analysis. However, its disadvantages include the need for centrifugation, which can cause lysis and loss of urinary cells, time-consuming, low efficiency, aggregation of urinary sediment after centrifugation, inability to form a single layer after spreading, and mixing of formed elements, seriously affecting the calculation, staining, and quantification of urinary sediment. Therefore, this method still relies on subjective experience for identification and semi-quantitative determination based on the number of particles per high-power field. The number of urine samples that each laboratory technician can accurately complete per day is small (n < 23 person-times), which cannot meet the urine test needs of a large number of outpatients. Since it is completed by clinical laboratory technicians through manual microscopy, it is more accurate than automatic analyzers. Therefore, manual microscopic examination is still the "gold standard" for urinary sediment examination.

[0031] Currently, for the problems existing in the related technologies, such as centrifugation damaging the morphology of urinary cells, overlapping of urinary sediment, difficulty in clustering, shedding of urinary cells during the staining process, numerous manual staining steps, and low efficiency of manual staining, no effective solutions have been proposed. Summary of the Invention

[0032] The object of the present invention is to provide a microfluidic chip, an intelligent staining detection device, and an intelligent staining method for detecting urinary sediment, aiming at the deficiencies in the existing technologies, to solve the problems such as centrifugation damaging the morphology of urinary cells, overlapping of urinary sediment, difficulty in clustering, shedding of urinary cells during the staining process, numerous manual staining steps, and low efficiency of manual staining.

[0033] To achieve the above object, the technical solution adopted by the present invention is:

[0034] In the first aspect, a microfluidic chip for detecting urinary sediment is provided, including:

[0035] An injection chamber for injecting urine and staining solution;

[0036] A separation chamber, which is arranged downstream of the injection chamber and is communicated with the injection chamber;

[0037] A microcolumn array, which is arranged in the separation chamber and is used for separating and clustering urinary sediment in urine;

[0038] A waste liquid chamber, which is arranged downstream of the separation chamber and is communicated with the separation chamber.

[0039] In some of the embodiments, the microcolumn array includes:

[0040] A number of microcolumns, and the number of microcolumns are arranged at intervals along the length direction and / or width direction of the separation chamber.

[0041] In some of the embodiments, in the microcolumn array, the density of the microcolumns increases from the upstream to the downstream of the separation chamber.

[0042] In some of these embodiments, in the microcolumn array, for several microcolumns in the same column, the distance between two adjacent microcolumns decreases from the upstream of the separation chamber to the downstream of the separation chamber.

[0043] In some of these embodiments, in the microcolumn array, for several microcolumns in two adjacent columns, the several microcolumns are arranged in an interleaved manner.

[0044] In some of these embodiments, the cross-section of the microcolumn is any one or a combination of several of a circle, an oval, a rhombus, a spindle shape, and an axisymmetric hexagon.

[0045] In some of these embodiments, the microcolumn array includes:

[0046] n microcolumn sub-arrays, the n microcolumn sub-arrays are sequentially arranged in the separation chamber, each microcolumn sub-array includes at least one microcolumn column, and each microcolumn column includes at least one microcolumn, and is used to separately separate n preset-size urinary sediments;

[0047] Wherein, the first preset size > the second preset size >... > the nth preset size.

[0048] In some of these embodiments, the microcolumn array includes:

[0049] A first microcolumn sub-array, the first microcolumn sub-array is arranged in the separation chamber and is used to separate urinary sediments of a first preset size;

[0050] A second microcolumn sub-array, the second microcolumn sub-array is arranged in the separation chamber and is located downstream of the first microcolumn sub-array, and is used to separate urinary sediments of a second preset size;

[0051] A third microcolumn sub-array, the third microcolumn sub-array is arranged in the separation chamber and is located downstream of the second microcolumn sub-array, and is used to separate urinary sediments of a third preset size;

[0052] Wherein, the first preset size > the second preset size > the third preset size.

[0053] In some of these embodiments, it further includes:

[0054] An information carrier, the information carrier is arranged on the microfluidic chip and is used to record the staining information of the microfluidic chip.

[0055] In a second aspect, there is provided an intelligent staining detection device, including:

[0056] A turntable unit, the turntable unit is rotatably arranged and is used to removably place at least one microfluidic chip as described in the first aspect;

[0057] An identification unit is arranged at the side of the turntable unit and is used for identifying the microfluidic chip to obtain information of the microfluidic chip.

[0058] A transfer unit is arranged at the side of the turntable unit and is used for transferring urine to the sample injection cavity of the microfluidic chip and transferring corresponding staining solution to the sample injection cavity of the microfluidic chip according to the information of the microfluidic chip.

[0059] A monitoring unit is arranged at the side of the turntable unit and is used for monitoring the staining processes of a plurality of the microfluidic chips.

[0060] A light source unit is arranged at the side of the turntable unit and is used for emitting a plurality of lights of different colors.

[0061] A filter unit is arranged at the side of the turntable unit and is used for filtering the light emitted by the light source unit.

[0062] A scanning unit is arranged at the side of the turntable unit and is used for acquiring image information of the microfluidic chip under the illumination of the light source unit and the filter unit.

[0063] A control unit is respectively connected to the turntable unit, the first transfer unit, the identification unit, the second transfer unit, the monitoring unit, the light source unit and the scanning unit.

[0064] In some embodiments, the light source unit includes at least four light sources.

[0065] In some embodiments, the filter unit includes at least four filter structures.

[0066] In some embodiments, it further includes:

[0067] A housing unit, wherein the turntable unit, the first transfer unit, the identification unit, the second transfer unit, the monitoring unit, the light source unit, the filter unit and the scanning unit are arranged inside the housing unit, the control unit is arranged at the side of the housing unit, and an insertion port and a discharge port are arranged at the side of the housing unit for inserting and discharging the microfluidic chip.

[0068] In a third aspect, an intelligent staining method is provided, which is applied to the intelligent staining detection device as described in the second aspect, and includes:

[0069] Transfer a preset volume of urine to the injection chamber of the microfluidic chip as described in the first aspect, then sequentially inject a preset volume of 95% ethanol and distilled water for rinsing, and finally dry it;

[0070] Transfer a preset volume of staining solution to the injection chamber of the microfluidic chip for staining, then inject a preset volume of distilled water for rinsing, and finally dry it;

[0071] Transfer a preset volume of 0.25% dilute hydrochloric acid to the injection chamber of the microfluidic chip to remove the excess staining solution, then inject a preset volume of distilled water for rinsing, and finally dry it;

[0072] Transfer a preset volume of dilute lithium carbonate solution to the injection chamber of the microfluidic chip for alkalization, then inject a preset volume of distilled water for rinsing, and finally dry it;

[0073] Transfer a preset volume of 95% ethanol to the injection chamber of the microfluidic chip for dehydration, then inject a preset volume of distilled water for rinsing, and finally dry it;

[0074] Transfer a preset volume of EA solution to the injection chamber of the microfluidic chip for staining;

[0075] Transfer a preset volume of 95% ethanol to the injection chamber of the microfluidic chip to remove the excess EA solution;

[0076] Transfer a preset volume of absolute ethanol to the injection chamber of the microfluidic chip for dehydration;

[0077] Transfer a preset volume of xylene to the injection chamber of the microfluidic chip for washing until it becomes transparent.

[0078] In some of these embodiments, it further includes:

[0079] Seal the slide with neutral balsam.

[0080] The present invention adopts the above technical solutions. Compared with the prior art, it has the following technical effects:

[0081] 1) Utilizing the hydrodynamic principle of urine particles, a microcolumn array is designed. Combined with the treatment of high adhesion and the movement through capillary channels, the fluid movement direction of urine sediment is controlled at the microscale through microcolumn arrays of different sizes and densities, separating urine sediment components with different particle sizes and morphologies, and realizing non-invasive "intelligent" separation of particles and formed components in urine liquid and their enrichment at different chip positions;

[0082] 2) Automatically realizes the enrichment and concentration of urinary cells in an ultra-large urine sample volume (urine sample greater than 100 ml), greatly improving the detection rate of microorganisms such as tumor cells and mycobacterium tuberculosis, and avoiding the situation that the detection results of small samples cannot represent the true urinary sediment characteristics of patients;

[0083] 3) The high-adhesion treatment of the urine sediment non-centrifugal microfluidic chip not only prevents the separation and shedding of urinary cells during staining, but also enables the thin-layer preparation of urine sediment cells, greatly reducing the overlap of formed elements separated, and improving the clarity of staining and the accuracy of subsequent artificial intelligence qualitative and quantitative analysis;

[0084] 4) Automatically separates and concentrates urine sediment components without centrifugation, ensuring the integrity of urinary cells and preventing damage to cells and other formed elements caused by urine centrifugation;

[0085] 5) Realizes the non-invasive "intelligent" separation of particles and formed elements in urine liquid and their enrichment at different chip positions in the microfluidic chip, which plays an important supporting role in improving the accuracy of urine sediment identification and diagnosis. Brief Description of the Drawings

[0086] Figure 1 is an exploded view of the microfluidic chip according to an embodiment of the present invention;

[0087] Figure 2 is a schematic diagram (one) of the fluid layer of the microfluidic chip according to an embodiment of the present invention;

[0088] Figure 3 is a schematic diagram (two) of the fluid layer of the microfluidic chip according to an embodiment of the present invention;

[0089] Figure 4 is a schematic diagram of the intelligent staining detection device according to an embodiment of the present invention;

[0090] Figure 5 is a circuit connection framework diagram of the intelligent staining detection device according to an embodiment of the present invention;

[0091] Figure 6 is a schematic diagram of the urine sediment staining monitoring method according to an embodiment of the present invention;

[0092] Figure 7 is a schematic diagram of the urine sediment panoramic scanning method according to an embodiment of the present invention;

[0093] Figure 8 is a schematic diagram of the urine sediment image segmentation and recognition method according to an embodiment of the present invention;

[0094] Figure 9 is a schematic diagram of the urine sediment image qualitative and quantitative analysis method according to an embodiment of the present invention, where a is the training flow chart and b is the test flow chart;

[0095] Figure 10 is a urinary sediment image according to an embodiment of the present invention;

[0096] Figure 11 is a urinary sediment image according to an embodiment of the present invention, wherein the urinary sediment podocalyxin staining: the cell nucleus is stained with Hoechst solution, and the urinary exfoliated cell podocalyxin staining positive cells [A], negative control [B] [[×200].

[0097] The reference numerals therein are: 100, microfluidic chip; 110, base layer; 111, first waste liquid hole; 120, fluid layer; 121, liquid inlet cavity; 122, separation cavity; 123, microcolumn array; 124, waste liquid cavity; 125, second waste liquid hole; 130, cover layer; 131, liquid inlet hole; 132, third waste liquid hole;

[0098] 200, intelligent staining detection device; 201, turntable unit; 202, recognition unit; 203, transfer unit; 204, monitoring unit; 205, light source unit; 206, filter unit; 207, scanning unit; 208, control unit; 209, housing unit. Detailed implementation manners

[0099] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0100] It should be noted that, without conflict, the embodiments in the present invention and the features in the embodiments may be combined with each other.

[0101] Next, the present invention will be further described in conjunction with the accompanying drawings and specific embodiments, but it is not a limitation of the present invention.

[0102] Embodiment 1

[0103] This embodiment relates to the microfluidic chip of the present invention.

[0104] A schematic embodiment of the present invention, as Figures 1 to 3 shown, a microfluidic chip 100 for detecting urinary sediment includes a base layer 110, a fluid layer 120 and a cover layer 130. Among them, the fluid layer 120 is disposed on the top of the base layer 110; the cover layer 130 is disposed on the top of the fluid layer 120.

[0105] The cross-section of the base layer 110 is rectangular.

[0106] The base layer 110 is made of a transparent material, including but not limited to one or more of glass, polydimethylsiloxane, polymethyl methacrylate, and polycarbonate, etc.

[0107] In some of the embodiments, the base layer 110 includes but is not limited to a substrate.

[0108] Furthermore, in one embodiment of the present invention, as Figure 1 shown, the base layer 110 includes a first waste liquid hole 111. Among them, the first waste liquid hole 111 is provided on the base layer 110 and is communicated with the fluid layer 120 for discharging waste liquid.

[0109] In the present invention, the first waste liquid hole 111 can be formed by laser etching, soft etching, photolithography, wet etching, or CNC precision machining.

[0110] The cross-section of the first waste liquid hole 111 is rectangular, waist-shaped, spindle-shaped, circular, trapezoidal, etc.

[0111] The size of the first waste liquid hole 111 matches the size of the base layer 110. Generally, the length of the first waste liquid hole 111 is less than the length of the base layer 110, the width of the first waste liquid hole 111 is less than the width of the base layer 110, and the depth of the first waste liquid hole 111 is equal to the thickness of the base layer 110.

[0112] The connection manner between the fluid layer 120 and the base layer 110 includes but is not limited to a fixed connection, such as integral molding, bonding, etc.

[0113] The cross-section of the fluid layer 120 is rectangular.

[0114] The size of the fluid layer 120 matches the size of the base layer 110. Generally, the length of the fluid layer 120 is equal to the length of the base layer 110, and the width of the fluid layer 120 is equal to the width of the base layer 110.

[0115] The fluid layer 120 is made of a transparent material, including but not limited to one or more of glass, polydimethylsiloxane, polymethyl methacrylate, and polycarbonate, etc.

[0116] In some of the embodiments, the fluid layer 120 includes but is not limited to a fluid plate.

[0117] Furthermore, as Figures 1 to 3As shown, the fluid layer 120 includes a liquid inlet chamber 121, a separation chamber 122, a microcolumn array 123, and a waste liquid chamber 124. Among them, the liquid inlet chamber 121 is arranged in the fluid layer 120 and is used for injecting urine and staining solution; the separation chamber 122 is arranged downstream of the sample injection chamber 121 and is communicated with the sample injection chamber 121; the microcolumn array 123 is arranged in the separation chamber 122 and is used for separating and clustering urinary sediment in urine; the waste liquid chamber 124 is arranged downstream of the separation chamber 122 and is communicated with the separation chamber 122..

[0118] In the present invention, there is no need to centrifuge the urine, avoiding the destruction of some cells in the urine during centrifugation. The microcolumn array 123 can separate and cluster the cells in the urine according to their sizes (similar to filtration by a sieve or a filter mesh), avoiding cell damage.

[0119] In the present invention, the flow rate of the urine is 0.28 - 1.5 m / s.

[0120] In the present invention, the liquid inlet chamber 121, the separation chamber 122, and the waste liquid chamber 124 can be formed by laser etching, soft etching, photolithography, wet etching, or CNC precision machining.

[0121] The cross-section of the liquid inlet chamber 121 is circular.

[0122] The size of the liquid inlet chamber 121 matches the size of the fluid layer 120. Generally, the radial size of the liquid inlet chamber 121 is smaller than the length and width of the fluid layer 120, and the depth of the liquid inlet chamber 121 is not greater than the thickness of the fluid layer 120.

[0123] In some embodiments, the liquid inlet chamber 121 penetrates through the fluid layer 120.

[0124] The cross-section of the separation chamber 122 is rectangular, waist-shaped, spindle-shaped, etc.

[0125] The size of the separation chamber 122 matches the size of the fluid layer 120. Generally, the length of the separation chamber 122 is smaller than the length of the fluid layer 120, the width of the separation chamber 122 is smaller than the width of the fluid layer 120, and the depth of the separation chamber 122 is not greater than the thickness of the fluid layer 120.

[0126] In some embodiments, the separation chamber 122 penetrates through the fluid layer 120.

[0127] In some embodiments, such as Figures 2 to 3As shown, the width of the separation chamber 122 shows an increasing, constant, and then decreasing trend from its upstream to its downstream. Specifically, the separation chamber 122 includes an inlet section, a separation section, and an outlet section. Among them, the inlet section is arranged downstream of the liquid inlet chamber 121 and is in communication with the liquid inlet chamber 121; the separation section is arranged downstream of the inlet section and is in communication with the inlet section, and a micro-column array 123 is arranged inside the separation section; the outlet section is arranged downstream of the separation section and upstream of the waste liquid chamber 124, and is in communication with the separation section and the waste liquid chamber 124 respectively.

[0128] The cross-section of the inlet section is an isosceles triangle, an isosceles trapezoid, a semi-circle, etc. Specifically, the width of the inlet section increases from its upstream to its downstream.

[0129] The cross-section of the separation section is a rectangle, an ellipse, a polygon, etc.

[0130] The size of the separation section matches the size of the inlet section. Generally, the width of the upstream end of the separation section is equal to the maximum width of the inlet section, and the length of the separation section is greater than the length of the inlet section.

[0131] The cross-section of the outlet section is an isosceles triangle, an isosceles trapezoid, a semi-circle, etc. Specifically, the width of the outlet section decreases from its upstream to its downstream.

[0132] The size of the outlet section matches the size of the separation section. Generally, the maximum width of the outlet section is equal to the width of the downstream end of the separation section, and the length of the outlet section is less than the length of the separation section.

[0133] The size of the outlet section matches the size of the inlet section. Generally, the maximum width of the outlet section is equal to the maximum width of the inlet section, the minimum width of the outlet section is equal to the minimum width of the inlet section, and the length of the outlet section is equal to the length of the inlet section.

[0134] Furthermore, the separation chamber 122 further includes an input channel and an output channel. Among them, the input channel is arranged downstream of the liquid inlet chamber 121 and upstream of the inlet section, and is in communication with the liquid inlet chamber 121 and the inlet section respectively; the output channel is arranged downstream of the outlet section and upstream of the waste liquid chamber 124, and is in communication with the outlet section and the waste liquid chamber 124 respectively.

[0135] The cross-section of the input channel is a rectangle.

[0136] The size of the input channel matches the size of the liquid inlet chamber 121. Generally, the width of the input channel is less than the radial dimension of the liquid inlet chamber 121, and the length of the input channel is greater than the radial dimension of the liquid inlet chamber 121.

[0137] The size of the input channel matches the size of the inlet section. Generally, the width of the input channel is not greater than the minimum width of the inlet section.

[0138] The cross-section of the output channel is a rectangle.

[0139] The size of the output channel matches the size of the outlet section. Generally, the width of the output channel is not greater than the minimum width of the outlet section.

[0140] For the microcolumn array 123 of the present invention, it is treated with a poly-D-lysine solution to achieve high adhesion.

[0141] In some embodiments thereof, the microcolumn array 123 is treated with a 0.05 - 0.20 mg / ml poly-D-lysine solution.

[0142] Furthermore, as Figures 1 to 2 shown, the microcolumn array 123 includes a plurality of microcolumns. Among them, the plurality of microcolumns are arranged at intervals along the length direction and / or the width direction of the separation chamber 122.

[0143] Generally, the plurality of microcolumns are arranged in an array inside the separation chamber 122. Specifically, the plurality of microcolumns form a plurality of microcolumn rows, and each microcolumn row includes at least one microcolumn.

[0144] For the plurality of microcolumn rows, the number of microcolumns in each microcolumn row may be the same or different. When the number of microcolumns in each microcolumn row is the same, the distance between two adjacent microcolumns in each microcolumn row may be the same or different. When the number of microcolumns in each microcolumn row is different, the distance between two adjacent microcolumns in each microcolumn row may be the same or different.

[0145] In some embodiments thereof, in the microcolumn array 123, the density of the microcolumns increases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. Specifically, the number of microcolumns in each microcolumn row increases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. For example, if there are n microcolumn rows, the first microcolumn row has a microcolumns, the second microcolumn row has a + 1 microcolumns, and so on, the nth microcolumn row has a + n - 1 microcolumns. Or, if there are n microcolumn rows, the first to the third microcolumn rows have a microcolumns, the fourth to the sixth microcolumn rows have a + 1 microcolumns, and so on, the (n - 2)th to the nth microcolumn rows have a + n / 3 - 1 microcolumns. That is, the number of microcolumns in each microcolumn row is different, or the number of microcolumns in some microcolumn rows is the same.

[0146] In some embodiments thereof, in the microcolumn array 123, for several microcolumns in the same row, the distance between two adjacent microcolumns decreases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. For example, if there are n microcolumn rows, the distance between two adjacent microcolumns in the first microcolumn row is d1, the distance between two adjacent microcolumns in the second microcolumn row is d2, and so on, the distance between two adjacent microcolumns in the nth microcolumn row is d n where, dn <... < d2 < d1. Alternatively, if there are n columns of micro-columns, the distance between adjacent micro-columns in the first to third columns of micro-columns is d1, the distance between adjacent micro-columns in the fourth to sixth columns of micro-columns is d2, and so on. The distance between adjacent micro-columns in the (n - 2)th to nth columns of micro-columns is d n / 3 , where d n / 3 <... < d2 < d1. That is, the distance between adjacent micro-columns in each micro-column array is different, or the distance between adjacent micro-columns in some micro-column arrays is the same.

[0147] For several micro-column arrays, the radial dimensions of the micro-columns in each micro-column array can be the same or different.

[0148] In some embodiments, in the micro-column array 123, the radial dimension of the micro-columns increases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. Specifically, the radial dimension of the micro-columns in each micro-column array increases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. For example, if there are n columns of micro-columns, the radial dimension of the micro-columns in the first column of micro-columns is R1, the radial dimension of the micro-columns in the second column of micro-columns is R2, and so on. The radial dimension of the micro-columns in the nth column of micro-columns is R n , where R1 < R2 <... < R n Or, if there are n columns of micro-columns, the radial dimension of the micro-columns in the first to third columns of micro-columns is R1, the radial dimension of the micro-columns in the fourth to sixth columns of micro-columns is R2, and so on. The radial dimension of the micro-columns in the (n - 2)th to nth columns of micro-columns is R n / 3 , where R1 < R2 <... < R n / 3 .

[0149] In a preferred embodiment of the present invention, the number of micro-columns in each micro-column array increases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122, and the distance between adjacent micro-columns in each micro-column array decreases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. In this way, urinary sediments of different sizes (generally referring to particle sizes) in urine can be separated and clustered. Specifically, the size distribution of urinary sediments decreases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. That is, the larger the size of the urinary sediment, the closer it is to the inlet section of the separation chamber 122, and the smaller the size of the urinary sediment, the closer it is to the outlet section of the separation chamber 122.

[0150] For several micro-column arrays, the distance between adjacent micro-column arrays can be the same or different..

[0151] In some of these embodiments, in the microcolumn array 123, the distance between two adjacent microcolumn columns decreases from the upstream of the separation chamber 122 to the downstream of the separation chamber 122. For example, if there are n microcolumn columns, the distance between the first microcolumn column and the second microcolumn column is d1, the distance between the second microcolumn column and the third microcolumn column is d2, and so on, the distance between the (n - 1)th microcolumn column and the nth microcolumn column is d n-1 , where d n-1 <... < d2 < d1. Alternatively, if there are n microcolumn columns, the distance between two adjacent microcolumns of the first to the third microcolumn columns is d1, the distance between two adjacent microcolumns of the third to the fifth microcolumn columns is d2, and so on, the distance between two adjacent microcolumns of the (n - 2)th to the nth microcolumn columns is d (n - 1) / 2,, where d (n-1) / 2 <... < d2 < d1. That is, the distances between two adjacent microcolumn columns are all different, or the distances between some adjacent microcolumn columns are the same.

[0152] In some of these embodiments, in the microcolumn array 123, for several microcolumns of two adjacent columns, the several microcolumns are arranged staggeredly. By using this arrangement, the flow distance of urine can be extended, urine deposition at a single position can be avoided, and the separation and clustering effect can be improved.

[0153] In some of these embodiments, the cross-section of the microcolumn is any one or a combination of several of a circle, an oval, a rhombus, a spindle shape, and an axisymmetric hexagon.

[0154] In some of these embodiments, the width of the end of the microcolumn facing the urine increases from the front end of the microcolumn to the rear end of the microcolumn. That is, the end of the microcolumn facing the urine is arc-shaped or pointed. Using this shape can divert urine and avoid urine deposition at a single position.

[0155] In some of these embodiments, the microcolumn array 123 includes n microcolumn sub-arrays (n ≥ 2). Among them, the n microcolumn sub-arrays are arranged in sequence in the separation chamber. Each microcolumn sub-column includes at least one microcolumn column, and each microcolumn column includes at least one microcolumn, which are used to separately separate n preset-sized urinary sediments.

[0156] Among them, the first preset size > the second preset size >... > the nth preset size.

[0157] In some of these embodiments, the micro-column array 123 includes a first micro-column sub-array, a second micro-column sub-array, and a third micro-column sub-array. Among them, the first micro-column sub-array is disposed in the separation chamber 122 and is used to separate urinary sediment of a first preset size; the second micro-column sub-array is disposed in the separation chamber 122 and is located downstream of the first micro-column sub-array and is used to separate urinary sediment of a second preset size; the third micro-column sub-array is disposed in the separation chamber 122 and is located downstream of the second micro-column sub-array and is used to separate urinary sediment of a third preset size.

[0158] Among them, the first preset size > the second preset size > the third preset size.

[0159] Generally, the number of micro-column rows of the first micro-column sub-array ≤ the number of micro-column rows of the second micro-column sub-array ≤ the number of micro-column rows of the third micro-column sub-array.

[0160] Generally, the number of micro-columns in each micro-column row of the first micro-column sub-array < the number of micro-columns in each micro-column row of the second micro-column sub-array < the number of micro-columns in each micro-column row of the third micro-column sub-array.

[0161] Generally, the distance between two adjacent micro-columns in each micro-column row of the first micro-column sub-array > the distance between two adjacent micro-columns in each micro-column row of the second micro-column sub-array > the distance between two adjacent micro-columns in each micro-column row of the third micro-column sub-array.

[0162] Generally, the distance between two adjacent micro-column rows of the first micro-column sub-array ≥ the distance between two adjacent micro-column rows of the second micro-column sub-array ≥ the distance between two adjacent micro-column rows of the third micro-column sub-array.

[0163] The cross-section of the waste liquid chamber 124 is rectangular, waist-shaped, spindle-shaped, circular, trapezoidal, etc.

[0164] The size of the waste liquid chamber 124 matches the size of the fluid layer 120. Generally, the length of the waste liquid chamber 124 is less than the length of the fluid layer 120, the width of the waste liquid chamber 124 is less than the width of the fluid layer 120, and the depth of the waste liquid chamber 124 is not greater than the thickness of the fluid layer 120.

[0165] The size of the waste liquid chamber 124 matches the size of the separation chamber 122. Generally, the minimum width of the waste liquid chamber 124 is greater than the width of the output channel.

[0166] The size of the waste liquid chamber 124 matches the size of the first waste liquid hole 111. Generally, the length of the waste liquid chamber 124 is not less than the length of the first waste liquid hole 111, and the width of the waste liquid chamber 124 is not less than the width of the first waste liquid hole 111.

[0167] In some of these embodiments, the width of the waste liquid chamber 124 increases from the upstream to the downstream of the waste liquid chamber 124. With this structure, the flow rate of the liquid can be slowed down, and the impact force of the liquid can be reduced, so that the liquid stays in the waste liquid chamber 124.

[0168] In some of these embodiments, the waste liquid chamber 124 is arranged through the fluid layer 120.

[0169] Furthermore, in an embodiment of the present invention, the fluid layer 120 further includes a second waste liquid hole 125. Wherein, the second waste liquid hole 125 is arranged through the side of the fluid layer 120 and communicates with the waste liquid chamber 124 for discharging waste liquid.

[0170] In the present invention, the second waste liquid hole 125 can be formed by laser etching, soft etching, photolithography, wet etching or CNC precision machining.

[0171] The cross-section of the second waste liquid hole 125 is rectangular, waist-shaped, spindle-shaped, circular, trapezoidal, etc.

[0172] The size of the second waste liquid hole 125 matches the size of the fluid layer 120. Generally, the length of the second waste liquid hole 125 is less than the length of the fluid layer 120, and the width of the second waste liquid hole 125 is less than the width of the fluid layer 120.

[0173] The size of the second waste liquid hole 125 matches the size of the waste liquid chamber 124. Generally, the length of the second waste liquid hole 125 is not greater than the length of the waste liquid chamber 124, and the width of the second waste liquid hole 125 is not greater than the width of the waste liquid chamber 124.

[0174] The connection manner between the cover layer 130 and the fluid layer 120 includes but is not limited to fixed connection, such as integral molding, bonding, etc.

[0175] The cross-section of the cover layer 130 is rectangular.

[0176] The size of the cover layer 130 matches the size of the fluid layer 120. Generally, the length of the cover layer 130 is equal to the length of the fluid layer 120, and the width of the cover layer 130 is equal to the width of the fluid layer 120.

[0177] The cover layer 130 is made of a transparent material, including but not limited to one or more of glass, polydimethylsiloxane, polymethyl methacrylate and polycarbonate processed, etc.

[0178] In some of these embodiments, the cover layer 130 includes but is not limited to a cover plate.

[0179] Furthermore, as Figure 1As shown, the cover layer 130 includes a liquid inlet hole 131. Among them, the liquid inlet cavity 121 is provided in the cover layer 130 and communicates with the liquid inlet cavity 121 of the fluid layer 120, and is used for injecting urine and staining liquid.

[0180] In the present invention, the liquid inlet hole 131 can be formed by laser etching, soft etching, photolithography, wet etching or CNC precision machining.

[0181] The cross-section of the liquid inlet hole 131 is circular.

[0182] The size of the liquid inlet hole 131 matches the size of the cover layer 130. Generally, the radial dimension of the liquid inlet hole 131 is smaller than the length and width of the cover layer 130, and the depth of the liquid inlet hole 131 is equal to the thickness of the cover layer 130.

[0183] The size of the liquid inlet hole 131 matches the size of the liquid inlet cavity 121. Generally, the radial dimension of the liquid inlet hole 131 is not greater than the radial dimension of the liquid inlet cavity 121.

[0184] Further, in an embodiment of the present invention, the cover layer 130 further includes a third waste liquid hole 132. Among them, the third waste liquid hole 132 is provided in the cover layer 130 and communicates with the waste liquid cavity 124 of the fluid layer 120, and is used for discharging waste liquid.

[0185] In the present invention, the third waste liquid hole 132 can be formed by laser etching, soft etching, photolithography, wet etching or CNC precision machining.

[0186] The cross-section of the third waste liquid hole 132 is rectangular, waist-shaped, spindle-shaped, circular, trapezoidal, etc.

[0187] The size of the third waste liquid hole 132 matches the size of the cover layer 130. Generally, the length of the third waste liquid hole 132 is smaller than the length of the cover layer 130, the width of the third waste liquid hole 132 is smaller than the width of the cover layer 130, and the depth of the third waste liquid hole 132 is equal to the thickness of the cover layer 130.

[0188] The size of the third waste liquid hole 132 matches the size of the waste liquid cavity 124. Generally, the length of the third waste liquid hole 132 is not greater than the length of the waste liquid cavity 124, and the width of the third waste liquid hole 132 is not greater than the width of the waste liquid cavity 124.

[0189] In the present invention, the first waste liquid hole 111, the second waste liquid hole 125, and the third waste liquid hole 132 can exist simultaneously or only one of them can exist. That is, there are the following several ways:

[0190] 1) Set the first waste liquid hole 111, and do not set the second waste liquid hole 125 and the third waste liquid hole 132;

[0191] 2) A second waste liquid hole 125 is provided, while the first waste liquid hole 111 and the third waste liquid hole 132 are not provided;

[0192] 3) A third waste liquid hole 132 is provided, while the first waste liquid hole 111 and the second waste liquid hole 125 are not provided;

[0193] 4) The first waste liquid hole 111 and the second waste liquid hole 125 are provided; the third waste liquid hole 132 is not provided;

[0194] 5) The first waste liquid hole 111 and the third waste liquid hole 132 are provided, while the second waste liquid hole 125 is not provided;

[0195] 6) The second waste liquid hole 125 and the third waste liquid hole 132 are provided, while the first waste liquid hole 111 is not provided;

[0196] 7) The first waste liquid hole 111, the second waste liquid hole 125 and the third waste liquid hole 132 are provided simultaneously.

[0197] Furthermore, the microfluidic chip 100 further includes an information carrier. The information carrier is disposed on the microfluidic chip 100 and is used to record the staining information of the microfluidic chip 100.

[0198] The staining information specifically refers to the staining method corresponding to the microfluidic chip 100, such as staining solution, etc.

[0199] Generally, the information carrier is disposed on the base layer 110 or the cover layer 130.

[0200] In some of these embodiments, the information carrier includes, but is not limited to, two-dimensional codes, barcodes, etc.

[0201] The usage method of the present invention is as follows:

[0202] Inject a preset volume of urine from the liquid inlet hole 131 into the liquid inlet cavity 121;

[0203] The urine flows from the liquid inlet cavity 121 to the separation cavity 122;

[0204] In the separation cavity 122, the urine is separated and clustered by the microcolumn array 123, and urine sediments of different sizes are distributed in the separation cavity 122. The waste urine flows into the waste liquid cavity 124 and is pumped out;

[0205] Inject a preset volume of staining solution from the liquid inlet hole 131 into the liquid inlet cavity 121 for staining, then inject a preset volume of distilled water from the liquid inlet hole 131 into the liquid inlet cavity 121 for rinsing. The waste liquid flows into the waste liquid cavity 124 and is pumped out, and finally dried (such as blown dry);

[0206] Inject 0.25% dilute hydrochloric acid of a preset volume from the liquid inlet hole 131 into the liquid inlet chamber 121 to remove the excess staining solution, then inject distilled water of a preset volume from the liquid inlet hole 131 into the liquid inlet chamber 121 for rinsing. The waste liquid flows into the waste liquid chamber 124 and is pumped out, and finally dried (such as by blowing dry);

[0207] Inject a preset volume of dilute lithium carbonate solution from the liquid inlet hole 131 into the liquid inlet chamber 121 for alkalization, then inject distilled water of a preset volume from the liquid inlet hole 131 into the liquid inlet chamber 121 for rinsing. The waste liquid flows into the waste liquid chamber 124 and is pumped out, and finally dried (such as by blowing dry);

[0208] Inject 95% ethanol of a preset volume from the liquid inlet hole 131 into the liquid inlet chamber 121 for dehydration, then inject distilled water of a preset volume from the liquid inlet hole 131 into the liquid inlet chamber 121 for rinsing. The waste liquid flows into the waste liquid chamber 124 and is pumped out, and finally dried (such as by blowing dry);

[0209] Inject a preset volume of EA solution from the liquid inlet hole 131 into the liquid inlet chamber 121 for staining, and the waste liquid flows into the waste liquid chamber 124 and is pumped out;

[0210] Inject 95% ethanol of a preset volume from the liquid inlet hole 131 into the liquid inlet chamber 121 to remove the excess EA solution, and the waste liquid flows into the waste liquid chamber 124 and is pumped out;

[0211] Inject absolute ethanol of a preset volume from the liquid inlet hole 131 into the liquid inlet chamber 121 for dehydration, and the waste liquid flows into the waste liquid chamber 124 and is pumped out;

[0212] Inject a preset volume of xylene from the liquid inlet hole 131 into the liquid inlet chamber 121 for washing until it becomes transparent, and the waste liquid flows into the waste liquid chamber 124 and is pumped out.

[0213] The technical effects of the present invention are as follows:

[0214] 1) Utilizing the hydrodynamic principle of urine particles, a microcolumn array is designed. Combined with the treatment of high adhesion and moving through capillary channels, the fluid movement direction of urine sediment is controlled at the microscale through microcolumn arrays of different sizes and densities, separating urine sediment components of different particle sizes and morphologies, and realizing non-invasive "intelligent" separation of particles and formed components in urine liquid and enriching them at different chip positions;

[0215] 2) Automatically realizing the enrichment and concentration of urine cells in an ultra-large urine sample volume (urine sample greater than 100 ml), greatly improving the detection rate of microorganisms such as tumor cells and mycobacterium tuberculosis, and avoiding the situation that the detection results of small samples cannot represent the true urine sediment characteristics of patients;

[0216] 3) The treatment of high adhesion of the urine sediment non-centrifugal microfluidic chip not only prevents the separation and shedding of urine cells during staining, but also enables the thin-layer preparation of urine sediment cells, greatly reducing the overlap of the separated formed components and improving the clarity of staining and the accuracy of subsequent artificial intelligence qualitative and quantitative analysis;

[0217] 4) Automatically separating and concentrating urine sediment components without centrifugation ensures the integrity of urine cells and prevents damage to cells and other formed components caused by urine centrifugation;

[0218] 5) Achieving non-invasive "intelligent" separation of particles and formed components in urine liquid and enriching them at different chip positions of the microfluidic chip plays an important supporting role in improving the accuracy of urine sediment identification and diagnosis.

[0219] Example 2

[0220] This example relates to the intelligent staining detection device of the present invention.

[0221] A schematic embodiment of the present invention, as Figures 4 to 5 shown, an intelligent staining detection device 200 includes a turntable unit 201, an identification unit 202, a transfer unit 203, a monitoring unit 204, a light source unit 205, a filter unit 206, a scanning unit 207 and a control unit 208. Among them, the turntable unit 201 is rotatably arranged for removably placing at least one microfluidic chip 100 as described in Example 1; the identification unit 202 is arranged on the side of the turntable unit 201 for identifying the microfluidic chip to obtain information of the microfluidic chip; the transfer unit 203 is arranged on the side of the turntable unit 201 for transferring urine to the sample injection cavity 121 of the microfluidic chip 100 and transferring the corresponding staining solution to the sample injection cavity 121 of the microfluidic chip 100 according to the information of the microfluidic chip 100; the monitoring unit 204 is arranged on the side of the turntable unit 201 for monitoring the staining process of several microfluidic chips 100; the light source unit 205 is arranged on the side of the turntable unit 201 for emitting several lights of different colors; the filter unit 206 is arranged on the side of the turntable unit 201 for filtering the light emitted by the light source unit 205; the scanning unit 207 is arranged on the side of the turntable unit 201 for obtaining the image information of the microfluidic chip 100 under the illumination of the light source unit 205 and the filter unit 206; the control unit 208 is respectively connected to the turntable unit 201, the first transfer unit 203, the identification unit 202, the second transfer unit 203, the monitoring unit 204, the light source unit 205 and the scanning unit 207.

[0222] Furthermore, the transfer unit 203 is also used for transferring auxiliary liquid to the sample injection cavity 121 of the microfluidic chip 100. Among them, the auxiliary liquid includes but is not limited to cleaning liquid, dehydrating liquid, etc.

[0223] In some of these embodiments, the turntable unit 201 includes a turntable carrier, a plurality of placement cavities, a rotating shaft, and a rotating motor. Among them, the turntable carrier is rotatably arranged; the plurality of placement cavities are distributed on the top of the turntable carrier and penetrate the turntable carrier respectively; the rotating shaft is arranged in the center of the turntable carrier for driving the turntable carrier to rotate; the rotating motor is connected to the rotating shaft and is connected to the control unit 208 for driving the rotating shaft to rotate.

[0224] The cross-section of the placement cavity is in an inverted convex shape. Specifically, the placement cavity is divided into an upper cavity and a lower cavity. Among them, the upper cavity penetrates the top of the turntable carrier and is used for placing the microfluidic chip 100; the lower cavity penetrates the bottom of the turntable carrier and is communicated with the upper cavity for allowing the light source to pass through.

[0225] The size of the upper cavity matches the size of the microfluidic chip 100. Generally, the length of the upper cavity is not less than the length of the microfluidic chip 100, and the width of the upper cavity is not less than the width of the microfluidic chip 100. Preferably, the length of the upper cavity is equal to the length of the microfluidic chip 100, and the width of the upper cavity is equal to the width of the microfluidic chip 100.

[0226] The size of the lower cavity matches the size of the upper cavity. Generally, the length of the lower cavity is less than the length of the upper cavity, the width of the lower cavity is less than the width of the upper cavity, and the thickness of the lower cavity is not greater than the thickness of the upper cavity.

[0227] In some of these embodiments, the identification unit 202 includes, but is not limited to, a barcode scanner for reading the information carrier located on the microfluidic chip 100.

[0228] In some of these embodiments, the transfer unit 203 includes a first multi-axis robotic arm and at least one pipetting structure. Among them, the first multi-axis robotic arm is arranged on the side of the turntable unit 201 and is connected to the control unit 208; the pipetting structure is arranged at the end of the first multi-axis robotic arm and is connected to the control unit 208 for sucking and discharging urine, staining solution, and other liquids.

[0229] In some of these embodiments, the first multi-axis robotic arm includes, but is not limited to, a Scara robot.

[0230] When there is one pipetting structure, each time the pipetting structure sucks a liquid, it needs to be cleaned with a cleaning solution before liquid transfer.

[0231] When there are multiple pipetting structures, a separate pipetting structure can be assigned to each type of liquid to avoid cross-contamination.

[0232] In some of these embodiments, the transfer unit 203 is arranged above the turntable unit 201.

[0233] In some of these embodiments, the monitoring unit 204 is disposed on the upper part of the turntable unit 201.

[0234] In some of these embodiments, the monitoring unit 204 includes, but is not limited to, an image collector, such as a video monitor.

[0235] In some of these embodiments, the light source unit 205 is disposed at the bottom of the turntable unit 201.

[0236] In some of these embodiments, the light source unit 205 includes at least four light sources. Specifically, the light source unit 205 includes at least a white light source, a red light source, a green light source, and a blue light source.

[0237] In some of these embodiments, the light source unit 205 includes 4 to 6 light sources.

[0238] In some of these embodiments, the filter unit 206 includes a filter carrier, a plurality of filter mounting cavities, and a plurality of filter plates. The filter carrier is disposed at the bottom of the turntable unit 201 and is configured to rotate with the turntable unit 201 or rotate independently; the plurality of filter mounting cavities are distributed on the filter carrier; and the plurality of filter plates are respectively disposed inside the corresponding filter mounting cavities for filtering light respectively.

[0239] Among them, the specifications of each filter mounting cavity are the same, that is, the shapes and sizes are the same.

[0240] Among them, the colors of the light that each filter plate can filter are different.

[0241] In some of these embodiments, the filter unit 206 includes at least four filter structures. Specifically, the filter unit 206 includes at least a white filter structure, a red filter structure, a green filter structure, and a blue filter structure.

[0242] In some of these embodiments, the filter unit 206 includes 4 to 6 filter structures.

[0243] In some of these embodiments, the scanning unit 207 includes a second multi-axis robotic arm and a scanning structure. Among them, the second multi-axis robotic arm is disposed on the side of the turntable unit 201 and is connected to the control unit 208; the scanning structure is disposed at the end of the second multi-axis robotic arm and is connected to the control unit 208 for scanning the microfluidic chip 100 to obtain image information of the microfluidic chip 100.

[0244] In some of these embodiments, the second multi-axis robotic arm includes, but is not limited to, a Scara robot.

[0245] In some of these embodiments, the scanning structure includes, but is not limited to, a high-speed camera, such as a high-speed fixed-focus camera (imaging frame rate ≥ 165fps).

[0246] In some of these embodiments, the control unit 208 includes, but is not limited to, a processor, a single-chip microcomputer, etc.

[0247] Furthermore, the intelligent staining detection device 200 further includes a housing unit 209. Among them, a turntable unit 201, a first transfer unit 203, an identification unit 202, a second transfer unit 203, a monitoring unit 204, a light source unit 205, a filter unit 206, and a scanning unit 207 are arranged inside the housing unit 209. A control unit 208 is arranged on the side of the housing unit 209. An insertion port and a discharge port are arranged on the side of the housing unit 209 for inserting and discharging the microfluidic chip 100.

[0248] Furthermore, the intelligent staining detection device 200 further includes a waste liquid recovery unit. Among them, the waste liquid recovery unit is arranged at the bottom of the turntable unit 201 and is communicated with the turntable unit 201 for recovering waste liquid.

[0249] In some of these embodiments, the waste liquid recovery unit includes a recovery box and a plurality of recovery channels. Among them, the recovery box is arranged at the bottom of the turntable unit 201; the plurality of recovery channels are telescopically distributed on the top of the recovery box and are respectively detachably connected to the corresponding placement cavities of the turntable unit 201 for recovering waste liquid.

[0250] Specifically, the plurality of recovery channels are respectively communicated with the waste liquid cavities 124 of the corresponding microfluidic chips 100 for recovering waste liquid.

[0251] More specifically, the plurality of recovery channels are respectively communicated with any one or several of the corresponding first waste liquid holes 111, second waste liquid holes, and third waste liquid holes 132.

[0252] Generally, the waste liquid in the waste liquid cavity 124 can flow to the corresponding recovery channel under the action of its own gravity.

[0253] In some of these embodiments, the waste liquid recovery unit further includes a plurality of pumps. Among them, the plurality of pumps are respectively communicated with the corresponding recovery channels for making the waste liquid in the waste liquid cavity 124 flow to the recovery channels.

[0254] In the present invention, the purpose of setting the pumps is to improve the discharge speed of the waste liquid.

[0255] The usage method of the present invention is as follows:

[0256] Put the microfluidic chip 100 into the housing unit 209 through the insertion port, and the microfluidic chip 100 is located in the placement cavity of the turntable unit 201;

[0257] The identification unit 202 identifies the staining information of the microfluidic chip 100 and transmits it to the control unit 208;

[0258] According to the staining information, the control unit 208 controls the transfer unit 203 to sequentially perform urine transfer, staining solution transfer, and auxiliary liquid transfer;

[0259] During the staining process of the microfluidic chip 100, the monitoring unit 204 performs real-time monitoring on the microfluidic chip 100 to determine whether the staining is normal;

[0260] After the staining is completed, the turntable unit 201 rotates to transfer the microfluidic chip 100 to the scanning position;

[0261] The control unit 208 controls the light source unit 205 to emit different light sources in a preset order and controls the filter unit 206 to rotate in a preset order, so that light of a preset color irradiates the microfluidic chip 100;

[0262] Meanwhile, the control unit 208 controls the scanning unit 207 to scan the microfluidic chip 100 in a preset order, thereby obtaining several pictures of the microfluidic chip 100 under the irradiation of light of different colors (generally, for the same microfluidic chip 100, the number of pictures obtained by the scanning unit 207 is a×b, where a is the number of lights and b is the number of pictures when each light irradiates);

[0263] The control unit 208 processes the pictures of several microfluidic chips 100 to obtain at least one urinary sediment image (generally, the number of urinary sediment images is 1 + a, where a is the number of lights);

[0264] By analyzing the urinary sediment image, the health status of the patient can be known.

[0265] Generally, the urinary sediment image includes:

[0266] 1) Simultaneously display urinary sediment images of different colors;

[0267] 2) Only display urinary sediment images of a single color.

[0268] The technical effects of the present invention are as follows:

[0269] 1) Automatic staining of the high-adhesion microcolumn array on the chip: The robotic arm automatically loads the sample and supports various urinary sediment stainings. It solves the problem of unattended operation, improves the detection efficiency, and provides technical support for formulating the standardization of urinary sediment analysis;

[0270] 2) Precise identification of the high-adhesion microcolumn array staining on the chip: The application of the separation technology of the non-centrifugal microfluidic chip for urinary sediment results in a preliminary classification of urinary sediment particles with similar sizes and charged formed components. Combining with urinary sediment staining, it greatly improves the accuracy of identifying urinary sediment formed components;

[0271] 3) Transparent treatment of the microfluidic tracks of the anti-fluorescence quenching mounting medium and gelatin glycerol mounting medium to achieve high-definition picture scanning and photography;

[0272] 4) Artificial intelligence recognition and quantitative analysis of the full-scan image of the original sample stained with high-adhesion microcolumn arrays on the microfluidic chip to achieve intelligent diagnosis of automatic urine cell recognition;

[0273] 5) Photography is carried out throughout the staining process to ensure that the entire flow process can be verified and permanent images can be saved;

[0274] 6) The non-centrifugal microfluidic chip for urine sediment is for single use, eliminating cross-contamination of samples;

[0275] 7) Due to low cost, simple instrument, and intelligent diagnosis, it is suitable for primary hospitals and community applications, benefiting a large number of patients.

[0276] Example 3

[0277] This example relates to the related method of the intelligent staining detection device of Example 2.

[0278] For the intelligent staining detection device of Example 2, it includes the following methods:

[0279] 1) Urine sediment staining monitoring method;

[0280] 2) Urine sediment panoramic scanning method;

[0281] 3) Urine sediment image segmentation and recognition method;

[0282] 4) Urine sediment image qualitative and quantitative analysis method.

[0283] As Figures 6 to 7 shown, for 1) the urine sediment staining monitoring method, it is specifically as follows:

[0284] First, divide the input stained image into multiple non-overlapping sub-blocks, and use five groups of convolutional layers to extract the convolutional features of each sub-block. Each group of convolutional layers contains two convolutional layers and one pooling layer. After five groups of convolutional layers, a multi-dimensional feature vector is obtained as the local feature of each sub-block. Then, use a weighted method to aggregate the local features, connect the aggregated features to a fully connected layer, use the average subjective score value of the image as the label, and perform supervised training to obtain a regression prediction model, so as to perform regression prediction on the image quality.

[0285] 1) Feature extraction

[0286] Input a color image with three RGB channels of size M×N. First, divide it into multiple sub-blocks of size 32×32×3. Use five groups of convolutional layers to extract the convolutional features of the sub-blocks. The first hidden layer contains two convolutional layers and one pooling layer. Convolve using 32 convolutional templates of size 3×3 to obtain features of size M×N×32. Downsample through the max pooling layer to obtain features as the input of the second hidden layer. Similarly, extract features for the second to fifth hidden layers respectively. Finally, after processing through five hidden layers, each sub-block is obtained as a 512-dimensional feature vector, which serves as the local feature of each sub-block. The ReLU is selected as the activation function for the convolutional layer, and only the threshold is required to obtain the activation value. Its calculation is as follows:

[0287] f(x) = max(0, x)

[0288] 2) Feature aggregation

[0289] To avoid the limitation of the image size, the method of cross-dimensional weighting can be used to aggregate the features. Thus, for images of different sizes, after feature extraction, features of the same dimension can be obtained.

[0290] First, calculate the spatial weights of each feature map. Suppose there are k channels, and each feature map is denoted as c k , with size W×H. Then the aggregation response at each spatial position (i, j) of all channels is Calculate the spatial weights of the feature map as follows:

[0291]

[0292] In the formula, both the values of α and β are 2, indicating l2 normalization processing.

[0293] Then, count the sparsity Ω of each channel, where λ (ij) is the channel vector.

[0294]

[0295] Thus, calculate the weight of each channel: where ε is a very small constant to ensure stability.

[0296] Multiply the extracted convolutional features by the spatial weights and sum them to obtain the spatially weighted features of the channels. Then multiply by the channel weights to obtain the k-dimensional aggregated features.

[0297] 3) Quality evaluation value

[0298] Through image convolution feature extraction and feature aggregation, a 512-dimensional feature vector can be obtained for each image. The feature vectors are connected to the first fully connected layer, passed through the Relu activation function, and then transmitted to the second fully connected layer. The second fully connected layer is connected to the output layer. Using the average subjective score value of the image as the label, supervised training is carried out, and the predicted quality evaluation value is directly output without passing through the activation function for non-linear transformation.

[0299] This algorithm is used to deeply evaluate the dyeing quality of the dyeing process and give feedback evaluation information. The evaluation information is transmitted to the sample motion control module through the video monitoring module, and the module directly drives the dyeing control motor to complete the intelligent control of the dyed sample and high-precision dyeing images.

[0300] Such as Figure 8 shown, for the 2) urinary sediment panoramic scanning method, it is as follows:

[0301] A multi-wavelength reading system is used to collect urinary sediment pictures, illuminated with white light and the three primary colors RGB, a total of 4 colors, and the image acquisition of 4-color pictures is completed. In the image acquisition, a high-speed camera combined with the movement of a two-dimensional plane scanning displacement platform is used for surface scanning shooting, and a method of completing the stitching of the full-frame image through local position features is adopted to construct a scanning and image analysis system for the entire panoramic dyeing image. Using a high-speed fixed-focus camera for imaging (imaging frame rate ≥ 165fps) solves the imaging quality problems such as trailing in high-speed moving imaging and improves the imaging speed of the system; using the high-precision Z-axis positioning ability of the Scara robot and combining the use of a fixed-focus microscope lens ensures the clarity of imaging.

[0302] Such as Figures 9a to 9b shown, for the 3) urinary sediment image segmentation and recognition method, it is as follows:

[0303] For the characteristics of urinary cells, when segmenting, not only the accurate position information of the cells but also the high-precision detailed information of the cells need to be obtained. Therefore, a deep network structure based on Unet is adopted, and the underlying information of the network is used to supplement the high-level information of the network, thus ensuring the accuracy of cell segmentation. Training of the U-net network: Based on the Caffe framework, the stochastic gradient descent method with momentum is adopted, and both the input image and its corresponding segmentation map are used for training. Without convolution, the output image is smaller than the input image. To reduce overhead and maximize the use of the GPU, larger input blocks with Overlap-tile are adopted. At the same time, a high momentum (0.99) is used, so that a large number of previously seen training samples can be used to determine the update in the current optimal step.

[0304] The energy function is calculated by combining the Soft-max value on the final feature map of the cross-entropy loss function. The calculation method of Soft-max is as follows:

[0305]

[0306] where α k (x) represents the activation value of the pixel at position X in the k-th layer of the feature map, where x ∈ Ω, k is the total number of classes of pixel points, and p k (x) is the approximate maximum function. The cross-entropy loss function is defined as follows:

[0307]

[0308] By pre-computing the weight map, the weight value of each pixel in the loss function is obtained, compensating for the different frequencies of each class of pixels in the training data, and making the network pay more attention to learning the edges between adjacent cells.

[0309]

[0310] where ω c (x) is the weight map for balancing class frequencies, d1(x) is the distance from this pixel point to the nearest cell boundary, and d2(x) is the distance from this pixel point to the second-nearest cell boundary.

[0311] For this training initialization, it is selected to follow a Gaussian distribution with a standard deviation of , where N represents the number of incoming nodes of a neuron. Since the training samples of urinary cells are relatively few and have large deformations, therefore, in the early stage of training, the samples are replicated with translational and rotational invariance and the gray value differences are adjusted to increase the sample data. At the same time, the samples are randomly deformed, and smooth deformations are generated on a rough 3×3 grid using random displacement vectors. The displacement vectors are sampled from a Gaussian distribution with a standard deviation of 10, and then bicubic interpolation is used to calculate the displacement of each pixel. In addition, Drop-out is added at the end of the contraction path to implicitly strengthen the sample data.

[0312] After implementing urinary cell image segmentation, the deep learning-based urinary cell image recognition method uses an improved generative adversarial network structure and a Soft-max classifier to achieve the recognition of urinary cell images. Utilizing the generative adversarial idea, within the framework of GAN, combining the Conditional Generative Adversarial Network (Conditional GAN, CGAN) and the Deep Convolutional Generative Adversarial Network (Deepconvolutional GAN, DCGAN), a Conditional Deep Convolutional Generative Adversarial Network model (Conditional-DCGAN, C-DCGAN) is established. The condition is added to the generator in the deep convolutional generative adversarial network. The generator G takes the random noise z and the label data c as inputs, connects them, and after linearization (Linear), three transposed convolutions, and activation by the hyperbolic tangent function (Tanh), outputs a one-dimensional tensor, which is a generated image sample. The discriminator D has a structure roughly opposite to that of the generator. It takes the image samples generated by the generator and the real urinary cell image samples as inputs, and after four convolutions, ReLU activation function, Meanpool average pooling, and linearization, obtains an evaluation value. Finally, in the fusion of the generator and the discriminator, the stochastic gradient descent algorithm is used to continuously iterate back and forth between the two to generate stable CDCGAN parameters. By leveraging the feature extraction ability of the convolutional network and supplemented by conditions for training, then the discriminator part in the trained C-DCGAN is extracted and Softmax is added to form the image recognition for urinary cells.

[0313] For the method of qualitative and quantitative analysis of 4) urinary sediment images, it is as follows:

[0314] The multi-wavelength image panoramic automatic scanning and reading control software completes the scanning of the slide sample by the microscopic CCD reading device to generate pictures, and transmits them to the PC together with the QR code information. The urinary sediment artificial intelligence recognition and quantitative analysis management software based on the standard urinary sediment color picture library for urinary system diseases and the urinary cell molecular recognition staining technology performs precise qualitative and quantitative analysis. The stained images are displayed on the computer screen under the video monitoring software. The urinary sediment analysis results are in the urine test item graphic report printing software, the patient medical record graphic database management software, and the image lossless compression remote transmission software, and finally a test report is generated.

[0315] Staining results: Epithelial cells: nuclei dark blue to purple blue, nucleoli red, cytoplasm pink to blue green; Red blood cells: bright red; White blood cells: cytoplasm light blue to green and nuclei dark blue black; Mucus: light blue to pink.

[0316] Example 4

[0317] This example relates to the intelligent staining method of the present invention.

[0318] A schematic embodiment of the present invention, an intelligent staining method, is applied to the intelligent staining detection device 200 described in Embodiment 2 or Embodiment 3, and includes:

[0319] Step S601: Transfer a preset volume of urine to the sample injection chamber 121 of the microfluidic chip 100 described in Embodiment 1, then sequentially inject a preset volume of 95% ethanol and distilled water for rinsing, and finally dry;

[0320] Step S602: Transfer a preset volume of staining solution to the sample injection chamber 121 of the microfluidic chip 100 for staining, then inject a preset volume of distilled water for rinsing, and finally dry;

[0321] Step S603: Transfer a preset volume of 0.25% dilute hydrochloric acid to the sample injection chamber 121 of the microfluidic chip 100 to remove the excess staining solution, then inject a preset volume of distilled water for rinsing, and finally dry;

[0322] Step S604: Transfer a preset volume of dilute lithium carbonate solution to the sample injection chamber 121 of the microfluidic chip 100 for alkalization, then inject a preset volume of distilled water for rinsing, and finally dry;

[0323] Step S605: Transfer a preset volume of 95% ethanol to the sample injection chamber 121 of the microfluidic chip 100 for dehydration, then inject a preset volume of distilled water for rinsing, and finally dry;

[0324] Step S606: Transfer a preset volume of EA solution to the sample injection chamber 121 of the microfluidic chip 100 for staining;

[0325] Step S607: Transfer a preset volume of 95% ethanol to the sample injection chamber 121 of the microfluidic chip 100 to remove the excess EA solution;

[0326] Step S608: Transfer a preset volume of absolute ethanol to the sample injection chamber 121 of the microfluidic chip 100 for dehydration;

[0327] Step S609: Transfer a preset volume of xylene to the sample injection chamber 121 of the microfluidic chip 100 for washing until it becomes transparent.

[0328] In step S601, the volume of urine is 0.1 ml to 200 ml.

[0329] In steps S601 to S605, drying includes but is not limited to blowing dry.

[0330] Further, after step S609, it further includes:

[0331] Step S610: Seal the slide with neutral balsam.

[0332] The technical effects of the present invention are basically the same as those of Embodiments 1 to 3, and will not be elaborated here.

[0333] Embodiment 5

[0334] This embodiment is a specific embodiment of Embodiment 4.

[0335] The patient in this embodiment is a patient with IgA nephritis, and the staining method is the modified Papanicolaou staining.

[0336] An intelligent staining method, comprising:

[0337] (1) Inject 1.0 ml of fresh urine into the sample pipeline of the high-adhesion urinary sediment microfluidic chip, then inject 0.3 ml of 95% ethanol (2 minutes), then inject 1.0 ml of distilled water, and dry the pipeline (1 minute);

[0338] (2) Inject hematoxylin staining solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip (12 minutes), take it out and inject distilled water for rinsing (1 minute), and dry the pipeline (1 minute);

[0339] (3) Put 0.25% dilute hydrochloric acid into the sample pipeline of the high-adhesion urinary sediment microfluidic chip to remove the excess hematoxylin staining solution (5 seconds), take it out and inject 2.0 ml of distilled water for rinsing (2 minutes), and dry the pipeline (2 minutes);

[0340] (4) Put dilute lithium carbonate solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for alkalization (0.5 minute), rinse with running water (2 minutes), and blow air on the pipeline (1 minute);

[0341] (5) Put 95% alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for dehydration (1 minute), rinse with running water (2 minutes), and dry the pipeline (1 minute);

[0342] (6) Put EA solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for staining (3 minutes);

[0343] (7) Put 95% alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip (5 seconds) to remove the excessive staining solution;

[0344] (8) Put 100% anhydrous alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for dehydration (5 seconds);

[0345] (9) Put xylene into the sample pipeline of the high-adhesion urinary sediment microfluidic chip and wash twice (10 seconds) for transparency;

[0346] (10) Put neutral balsam into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for mounting.

[0347] The stained image is as follows Figure 6 shown, and the analysis results are as follows:

[0348] 1) Epithelial cells: The nucleus is dark blue to violet blue, the nucleolus is red, and the cytoplasm is pink to blue-green;

[0349] 2) Red blood cells: Bright red;

[0350] 3) White blood cells: The cytoplasm is light blue to green and the nucleus is dark blue-black;

[0351] 4) Mucus: Light blue to pink;

[0352] 5) The casts are blue.

[0353] Example 6

[0354] This example is a specific example of Example 4.

[0355] The patient in this example is a bladder cancer patient, and the staining method is modified Papanicolaou staining.

[0356] An intelligent staining method includes:

[0357] (1) Inject 50.0 ml of fresh urine into the sample pipeline of the high-adhesion urinary sediment microfluidic chip, then inject 0.3 ml of 95% ethanol (2 minutes), then inject 1.0 ml of distilled water, and dry the pipeline (1 minute);

[0358] (2) Inject hematoxylin staining solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip (12 minutes), take it out and inject distilled water for rinsing (1 minute), and dry the pipeline (1 minute);

[0359] (3) Put 0.25% dilute hydrochloric acid into the sample pipeline of the high-adhesion urinary sediment microfluidic chip to remove the excess hematoxylin staining solution (5 seconds), take it out and inject 2.0 ml of distilled water for rinsing (2 minutes), and dry the pipeline (2 minutes);

[0360] (4) Put dilute lithium carbonate solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for alkalization (0.5 minute), rinse with running water (2 minutes), and blow dry the pipeline (1 minute);

[0361] (5) Put 95% alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for dehydration (1 minute), rinse with running water (2 minutes), and dry the pipeline (1 minute);

[0362] (6) Put EA solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for staining (3 minutes);

[0363] (7) Put 95% alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip (for 5 seconds) to remove excessive staining solution;

[0364] (8) Put 100% absolute alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for dehydration (for 5 seconds);

[0365] (9) Put xylene into the sample pipeline of the high-adhesion urinary sediment microfluidic chip and wash twice (for 10 seconds) to make it transparent;

[0366] (10) Seal the sample pipeline of the high-adhesion urinary sediment microfluidic chip with neutral gum.

[0367] The analysis results of the stained image are as follows:

[0368] The tumor cell nuclei are deeply stained, the cell volume is large, the morphology is uneven, and the cytoplasm is pink.

[0369] Example 7

[0370] This example is a specific example of Example 4.

[0371] The patient in this example is a diabetic patient, and the staining method is modified Papanicolaou staining.

[0372] An intelligent staining method includes:

[0373] (1) Inject 25.0 ml of fresh urine into the sample pipeline of the high-adhesion urinary sediment microfluidic chip, then inject 0.3 ml of 95% ethanol (for 2 minutes), then inject 1.0 ml of distilled water, and dry the pipeline (for 1 minute);

[0374] (2) Inject hematoxylin staining solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip (for 12 minutes), take it out and inject distilled water for rinsing (for 1 minute), and dry the pipeline (for 1 minute);

[0375] (3) Put 0.25% dilute hydrochloric acid into the sample pipeline of the high-adhesion urinary sediment microfluidic chip to remove the excess hematoxylin staining solution (for 5 seconds), take it out and inject 2.0 ml of distilled water for rinsing (for 2 minutes), and dry the pipeline (for 2 minutes);

[0376] (4) Put dilute lithium carbonate solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for alkalization (for 0.5 minute), rinse with running water (for 2 minutes), and blow dry the pipeline (for 1 minute);

[0377] (5) Put 95% alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for dehydration (for 1 minute), rinse with running water (for 2 minutes), and dry the pipeline (for 1 minute);

[0378] (6) Place the EA solution into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for staining (3 minutes);

[0379] (7) Place 95% alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip (5 seconds) to remove excessive staining solution;

[0380] (8) Place 100% absolute alcohol into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for dehydration (5 seconds);

[0381] (9) Place xylene into the sample pipeline of the high-adhesion urinary sediment microfluidic chip and wash twice (10 seconds) for clearing;

[0382] (10) Place neutral balsam into the sample pipeline of the high-adhesion urinary sediment microfluidic chip for mounting.

[0383] The stained image is as Figure 7 shown, and the analysis results are as follows:

[0384] The cell membrane of glomerular epithelial cells (podocytes) is stained yellow-brown, and the nucleus is blue.

[0385] The above are only the preferred embodiments of the present invention, and do not limit the implementation manners and protection scope of the present invention. For those skilled in the art, it should be realized that all equivalent substitutions and obvious changes made by using the description and illustrations of the present invention should be included in the protection scope of the present invention.

Claims

1. A microfluidic chip for detecting urine sediment, characterized in that: include: The sample injection chamber is used to inject urine and dye solution; A separation chamber, the separation chamber is arranged downstream of the injection chamber and communicated with the injection chamber; A micro-column array, which is arranged in the separation chamber and is used for separating and clustering urine sediment; A waste liquid chamber, wherein the waste liquid chamber is arranged downstream of the separation chamber and communicated with the separation chamber.

2. The microfluidic chip according to claim 1, characterized in that: The micro-pillar array comprises: A plurality of micro-pillars are arranged at intervals along the length direction and / or width direction of the separation chamber.

3. The microfluidic chip according to claim 2, characterized in that: In the micro-pillar array, the density of the micro-pillars increases from the upstream of the separation chamber to the downstream of the separation chamber; and / or In the micro-pillar array, for a plurality of the micro-pillars in the same column, the distance between two adjacent micro-pillars decreases from the upstream of the separation chamber to the downstream of the separation chamber; and / or In the micro-column array, for a plurality of the micro-columns in two adjacent columns, the plurality of the micro-columns are arranged in an interlaced manner; and / or The cross section of the microcolumn is any one of a circle, an oval, a rhombus, a spindle, and an axisymmetric hexagon, or a combination of the above.

4. The microfluidic chip according to claim 2 or 3, characterized in that: The micro-pillar array comprises: n micro-pillar arrays, the n micro-pillar arrays are sequentially arranged in the separation chamber, each of the micro-pillar arrays includes at least one micro-pillar column, and each of the micro-pillar columns includes at least one micro-pillar, for separating urine sediments of n preset sizes respectively; Among them, the first preset size>the second preset size>...>the nth preset size.

5. The microfluidic chip according to claim 4, characterized in that: The micro-pillar array comprises: A first micro-pillar array, which is disposed in the separation chamber and is used to separate urine sediment of a first preset size; A second micro-pillar array, which is disposed in the separation chamber and located downstream of the first micro-pillar array, and is used to separate urine sediment of a second preset size; a third micro-column array, the third micro-column array being disposed in the separation chamber and located downstream of the second micro-column array, and being used for separating urine sediment of a third preset size; Among them, the first preset size>the second preset size>the third preset size.

6. The microfluidic chip according to claim 1, characterized in that: Also includes: An information carrier is arranged on the microfluidic chip and is used to record the staining information of the microfluidic chip.

7. An intelligent dyeing detection device, characterized in that: include: A turntable unit, the turntable unit being rotatably arranged and used for removably placing at least one microfluidic chip according to any one of claims 1 to 6; an identification unit, the identification unit being disposed on a side of the turntable unit and being used for identifying the microfluidic chip to obtain information of the microfluidic chip; A transfer unit, which is disposed on the side of the turntable unit and is used to transfer urine to the injection cavity of the microfluidic chip and transfer corresponding dyeing liquid to the injection cavity of the microfluidic chip according to information of the microfluidic chip; A monitoring unit, which is disposed on the side of the turntable unit and is used to monitor the dyeing process of the microfluidic chips; A light source unit, the light source unit being disposed on a side of the turntable unit and configured to emit light of a plurality of different colors; A filter unit, the filter unit is arranged at a side of the turntable unit and is used to filter the light emitted by the light source unit; A scanning unit, which is disposed on a side of the turntable unit and is used to obtain image information of the microfluidic chip under the illumination of the light source unit and the filter unit; A control unit, wherein the control unit is respectively connected to the turntable unit, the first transfer unit, the identification unit, the second transfer unit, the monitoring unit, the light source unit, and the scanning unit.

8. The intelligent dyeing detection device according to claim 7, characterized in that: Also includes: A shell unit, wherein the turntable unit, the first transfer unit, the identification unit, the second transfer unit, the monitoring unit, the light source unit, the filter unit, and the scanning unit are arranged inside the shell unit, the control unit is arranged on the side of the shell unit, and the side of the shell unit is provided with an insertion port and a discharge port for inserting and discharging the microfluidic chip.

9. An intelligent dyeing method, applied to the intelligent dyeing detection device according to any one of claims 6 to 8, characterized in that: include: Transferring a preset volume of urine to the injection chamber of the microfluidic chip according to any one of claims 1 to 5, then injecting preset volumes of 95% ethanol and distilled water in sequence for rinsing, and finally drying; Transferring a preset volume of dye solution to the injection chamber of the microfluidic chip for dyeing, then injecting a preset volume of distilled water for rinsing, and finally drying; Transferring a preset volume of 0.25% dilute hydrochloric acid to the injection chamber of the microfluidic chip to remove excess staining solution, then injecting a preset volume of distilled water for rinsing, and finally drying; Transferring a preset volume of dilute lithium carbonate solution to the injection chamber of the microfluidic chip for alkalization, then injecting a preset volume of distilled water for rinsing, and finally drying; Transferring a preset volume of 95% ethanol to the injection chamber of the microfluidic chip for dehydration, then injecting a preset volume of distilled water for rinsing, and finally drying; Transferring a preset volume of EA solution to the injection chamber of the microfluidic chip for staining; Transferring a preset volume of 95% ethanol to the injection chamber of the microfluidic chip to remove excess EA liquid; Transferring a preset volume of anhydrous ethanol to the injection chamber of the microfluidic chip for dehydration; A preset volume of xylene is transferred to the injection chamber of the microfluidic chip for washing until it becomes transparent.

10. The intelligent dyeing method according to claim 9, characterized in that: Also includes: Use neutral gum to mount the slides.

Images