Sampling mechanism, cell enrichment device and method
The sampling mechanism controlled by negative and positive pressure simplifies the cell enrichment process, solves the problems of cumbersome procedures and low cell quantity sample enrichment in existing technologies, achieves efficient cell enrichment and uniform sedimentation, and improves the reliability of detection.
Patent Information
- Application Number
- CN202511617594.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-06
- Publication Date
- 2026-01-23
AI Technical Summary
In existing technologies, membrane filtration and centrifugation methods involve cumbersome steps in the cell enrichment process, and samples with low target cell content are difficult to enrich to a sufficient quantity for detection, which affects diagnostic efficiency.
A sampling mechanism is employed, comprising a membrane tube, a membrane tube coupler, and a filter membrane. The adsorption and detachment of target cells are achieved through negative and positive pressure control components, simplifying the operation process and increasing the sample volume.
It improves cell pretreatment efficiency, reduces the difficulty of enriching low-cell-volume samples, ensures the complete transfer and uniform sedimentation of target cells, and enhances the reliability and efficiency of detection.
Smart Images
Figure CN121384573A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of cell enrichment, and in particular to a sampling mechanism, a cell enrichment device and a method. BACKGROUND
[0002] Liquid-based cytology (LBC) is a diagnostic method for cytology, which involves the steps of preparing a cell sample, coating target cells on a glass slide for observation under a microscope. LBC places the sample in a cell preservative, suspends the target cells in the preservative, and then uses membrane filtration or centrifugation to remove background cells (such as blood cells) and enrich target cells. This process is called pre-preparation processing. The purified and enriched target cells are transferred or settled on a glass slide, which is called preparation. Enriching target cells can increase the number of target cells on the glass slide, thereby increasing the probability of detecting abnormal cells.
[0003] Currently, membrane filtration and centrifugation are commonly used to remove background cells and enrich target cells.
[0004] Membrane filtration is further divided into negative pressure filtration and positive pressure filtration. Negative pressure filtration is achieved by starting a negative pressure device, so that background cells are filtered out and individual larger target cells are retained on the surface of the filter membrane. However, it not only requires the step of turning the membrane cartridge, but also the process of transferring cells from the membrane to the glass slide can change the cell morphology (physical extrusion), thereby increasing the difficulty of diagnosis. Positive pressure filtration is achieved by inputting air into the filter membrane cartridge to filter out mucus, blood cells, etc., and then injecting buffer to resuspend the target cells on the surface of the filter membrane and transfer them to the glass slide. This method can only handle about 2 milliliters of sample liquid, which is not conducive to the enrichment of low cell volume samples. At the same time, during the resuspension process, part of the target cells remain on the surface of the filter membrane and cannot be completely transferred to the glass slide, which further increases the difficulty of enriching low cell volume samples.
[0005] Centrifugation requires two steps of centrifugation to remove background cells and collect target cells. The first step is performed in a preservative, which aims to remove part of the background cells and remove the preservative. The second step is performed in a gradient centrifugation liquid, which aims to further concentrate the target cells and remove blood cells. Centrifugation not only has a complex operation process, but also easily loses patient information during the entire sample transfer process. SUMMARY
[0006] The present application discloses a sampling mechanism, a cell enrichment device and a method to solve the technical problems of complicated steps and difficulty in enriching low target cell content samples to sufficient target cells for detection when using membrane filtration for cell preparation in the related art.
[0007] To solve the above problems, the technical scheme adopted by the present application is as follows: The first aspect of the present application provides a sampling mechanism.
[0008] The sampling mechanism of the application is used in the cell enrichment device, and comprises a membrane cylinder, a membrane cylinder coupler and a filter membrane.
[0009] The second aspect of the application provides a cell enrichment device.
[0010] The cell enrichment device of the application comprises a sampling mechanism, a sample mechanism and a sedimentation mechanism, wherein, The sampling mechanism is the sampling mechanism described in any of the technical solutions of the application. The sample mechanism is used for placing a sample cup. The sedimentation mechanism comprises a sedimentation bin and a glass slide.
[0011] The third aspect of the application provides a cell enrichment method.
[0012] The cell enrichment method of the application is implemented by using the cell enrichment device of any of the technical solutions of the application. The cup cover of the sample cup is unscrewed, the membrane cylinder coupler of the sampling mechanism is coupled with the membrane cylinder, and then the membrane cylinder is transferred to the sample cup with the cup cover unscrewed. The membrane cylinder is lowered to immerse the filter membrane in the sample liquid, a negative pressure pump is started to keep the membrane cylinder in negative pressure, and target cells are adsorbed on the filter membrane. After the suction is completed, the membrane cylinder is transferred to the sedimentation bin, the negative pressure pump is turned off, a positive pressure pump is started to keep the membrane cylinder in positive pressure, target cells are detached from the filter membrane and are settled on the glass slide.
[0013] The technical solution adopted by the application can achieve the following beneficial effects: The sampling mechanism of the application can make the target cells adsorbed on the distal end surface of the filter membrane while the sample liquid in the sample cup is pumped out in the pretreatment stage, and the membrane cylinder maintains negative pressure, compared with the prior art, not only the step of turning over the membrane cylinder to pump out the sample liquid can be saved, but also the structure of the cell enrichment device can be simplified; in addition, the sample liquid can be continuously pumped out by negative pressure, so that the amount of sample to be processed is greatly increased, for the sample with low content of target cells, by increasing the sample amount, enough target cells can be enriched for detection, the cell pretreatment efficiency and reliability are improved, and the diagnosis difficulty of low cell amount sample is solved.
[0014] The sampling mechanism of the application can make the target cells adsorbed on the distal end surface of the filter membrane while the sample liquid in the sample cup is pumped out in the pretreatment stage, and the membrane cylinder maintains negative pressure, compared with the prior art, not only the step of turning over the membrane cylinder to pump out the sample liquid can be saved, but also the structure of the cell enrichment device can be simplified; in addition, the sample liquid can be continuously pumped out by negative pressure, so that the amount of sample to be processed is greatly increased, for the sample with low content of target cells, by increasing the sample amount, enough target cells can be enriched for detection, the cell pretreatment efficiency and reliability are improved, and the diagnosis difficulty of low cell amount sample is solved. BRIEF DESCRIPTION OF DRAWINGS
[0015] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description are only some embodiments of the application, and other drawings can also be obtained by those skilled in the art without creative labor.
[0016] Figure 1 is the overall schematic diagram of the cell enrichment device of the embodiment of the application; Figure 2 is the first partial schematic diagram of the cell enrichment device of the embodiment of the application; Figure 3 is the second partial schematic diagram of the cell enrichment device of the embodiment of the application; Figure 4 is the partial schematic diagram of the sampling mechanism of the embodiment of the application; Figure 5 is Figure 4 is the enlarged view of part A in Figure 6 is the schematic diagram of the membrane cylinder of the embodiment of the application; Figure 7 is the schematic diagram of the membrane cylinder support of the embodiment of the application; Figure 8 is the schematic diagram of the sample conveying belt of the embodiment of the application; Figure 9 is the overall schematic diagram of the sedimentation mechanism of the embodiment of the application; Figure 10 is the partial schematic diagram of the sedimentation mechanism of the embodiment of the application; Figure 11 is Figure 10Enlarged view of part B; Figure 12 Figure is a schematic diagram of the sedimentation chamber of the embodiment of the present application.
[0017] In the figure: 100, sampling mechanism; 110, membrane cartridge; 111, convex edge; 112, first barrel; 113, second barrel; 114, connecting part; 120, membrane cartridge coupler; 121, stepped surface; 130, filter membrane; 141, first needle tube; 142, negative pressure pump; 143, second needle tube; 144, positive pressure pump; 151, third needle tube; 152, plunger pump; 160, pressure detection assembly; 170, first mechanical arm; 180, membrane cartridge support; 181, membrane cartridge scraper; 182, cleaning pool; 200, sample mechanism; 210, sample cup; 211, cup cover; 220, sample conveying belt; 230, cup taking mechanical gripper; 240, cap screwing assembly; 241, rotating mechanical gripper; 300, sedimentation mechanism; 310, sedimentation chamber; 320, glass slide; 330, sedimentation disc. DETAILED DESCRIPTION
[0018] In order to make the objectives, technical solutions and advantages of the present application clearer, the technical solutions of the present application will be described in detail below. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of protection of the present application.
[0019] The terms "first", "second", etc. in the specification and claims of the present application are used to distinguish similar objects, and are not used to describe a specific order or sequence. It should be understood that the data used in this way can be interchanged under appropriate circumstances, so that the embodiments of the present application can be implemented in an order other than those illustrated or described herein, and the objects distinguished by "first", "second", etc. are generally of a kind and are not limited to the number of objects, for example, the first object can be one or more. In addition, "and / or" in the specification and claims indicates at least one of the connected objects, and the character " / ", generally indicates that the front and rear associated objects are in an "or" relationship.
[0020] In the embodiments of the present application, the "proximal end face" and "distal end face" of the filter membrane refer to the relative position of the filter membrane to the membrane cartridge coupler in the use environment, wherein the end closer to the membrane cartridge coupler is designated as the "proximal end face", and the end farther from the membrane cartridge coupler is designated as the "distal end face".
[0021] The specific embodiments and application scenarios of the sampling mechanism, cell enrichment device and method provided by the present application will be described in detail below in conjunction with the accompanying Figures 1 to 12 , the specific embodiments and application scenarios of the sampling mechanism, cell enrichment device and method provided by the present application will be described in detail below in conjunction with the accompanying
[0022] The sampling mechanism of the embodiment is used in the cell enrichment device. Specifically, the cell enrichment device needs to go through at least the following processes to complete the enrichment of cells: first, the blood cells are removed and the target cells are enriched by membrane filtration method; and then the enriched cells are uniformly transferred on a slide. The sampling mechanism in the cell enrichment device is mainly involved in the steps of removing blood cells, enriching target cells, and uniformly transferring the enriched cells on a slide.
[0023] The sampling mechanism of the embodiment includes a membrane cylinder 110, as shown in Figures 4 to 6 The membrane cylinder 110 has an open structure at both ends. For example, the membrane cylinder 110 has a tubular structure with a height of 20-40 mm and an inner diameter of 8-15 mm. For example, the membrane cylinder 110 is integrally formed by injection molding, as shown in Figure 6 The membrane cylinder 110 is made of transparent plastic material.
[0024] The sampling mechanism of the embodiment further includes a membrane cylinder coupler 120, as shown in Figure 4 and Figure 5 The membrane cylinder coupler 120 is installed below the first mechanical arm 170 and can be moved by the movement of the first mechanical arm 170. For example, the first mechanical arm 170 can move in three directions XYZ in three-dimensional space. For example, the membrane cylinder coupler 120 is made of metal or other hard materials.
[0025] The sampling mechanism of the embodiment further includes a filter membrane 130, as shown in Figures 4 to 6 For example, the filter membrane 130 is composed of high molecular materials, and the filter membrane 130 is provided with filter holes with a pore size of 8-12 microns. The filter holes allow red blood cells, white blood cells, and mucus to pass through, while blocking the target cells from passing through. The target cells are epithelial cells or gland cells, which can also be referred to as cells to be detected.
[0026] In some embodiments, the membrane cylinder 110 has an open structure at both ends, and the filter membrane 130 is fixed to the opening at one end of the membrane cylinder 110. For example, the filter membrane 130 is fixed to the distal end face of the membrane cylinder 110, as shown in Figure 5 and Figure 6 For example, the filter membrane 130 is fixed to the distal end face of the membrane cylinder 110 by welding or bonding. The membrane cylinder coupler 120 can be coupled or decoupled with the other end of the membrane cylinder 110. For example, the membrane cylinder coupler 120 is inserted into the membrane cylinder 110 and tightly connected with the membrane cylinder 110, so as to realize the coupling of the membrane cylinder coupler 120 with the membrane cylinder 110, as shown in Figure 4 and Figure 5 The membrane cylinder coupler 120 is separated from the membrane cylinder 110, so as to realize the decoupling of the membrane cylinder coupler 120 with the membrane cylinder 110.
[0027] Exemplarily, the filter membrane 130 is a high-molecular membrane material, which has certain ductility. Thus, the filter membrane 130 can be deformed when subjected to external force, reducing the risk of damage to the filter membrane 130.
[0028] In the embodiment, the coupling refers to that the membrane cylinder coupler 120 is inserted into the membrane cylinder 110, and the membrane cylinder coupler 120 and the membrane cylinder 110 form a sealed state of close contact, and the membrane cylinder 110 can be driven to move synchronously by the movement of the membrane cylinder coupler 120.
[0029] In some embodiments, the membrane cylinder coupler 120 is integrated with a pressure control assembly. When the membrane cylinder coupler 120 is coupled with the membrane cylinder 110, the pressure control assembly is used to control the pressure in the membrane cylinder 110. Exemplarily, when the membrane cylinder coupler 120 is coupled with the membrane cylinder 110, the pressure control assembly is used to maintain negative pressure in the membrane cylinder 110 in the pretreatment stage, and is used to maintain positive pressure in the membrane cylinder 110 in the slice preparation stage. Specifically, in the pretreatment stage, maintaining negative pressure in the membrane cylinder 110 can make the target cells adsorbed to the distal end surface of the filter membrane 130, and at the same time, the sample liquid in the sample cup 210 can be pumped out. In the slice preparation stage, maintaining positive pressure in the membrane cylinder 110 can make the target cells adsorbed to the filter membrane 130 fall off, so as to facilitate the target cells to be settled on the slide.
[0030] In the pretreatment stage, the sampling mechanism of the embodiment can make the target cells adsorbed to the distal end surface of the filter membrane 130 by maintaining negative pressure in the membrane cylinder 110, and at the same time, the sample liquid in the sample cup 210 can be pumped out. Compared with the prior art, the sampling mechanism of the embodiment can not only omit the step of turning over the membrane cylinder to pump out the sample liquid, but also can simplify the structure of the cell enrichment device, and improve the cell pretreatment efficiency and reliability.
[0031] On the other hand, in the slice preparation stage, the sampling mechanism of the embodiment can make the target cells adsorbed to the filter membrane 130 fall off by maintaining positive pressure in the membrane cylinder 110, which can realize the full falling of the target cells on the filter membrane 130. For the slice preparation of a low-cell-quantity sample, the sampling mechanism of the embodiment can reduce the risk of loss of the target cells, thereby reducing the difficulty of enrichment of the low-cell-quantity sample.
[0032] In some embodiments, the pressure control assembly includes a first needle tube 141 and a negative pressure pump 142, as shown in FIG. 2. Figure 2 and Figure 5As shown. The first needle tube 141 is in communication with the negative pressure pump 142. Exemplarily, the first needle tube 141 is in communication with the negative pressure pump 142 through a hose or a hard tube. When the negative pressure pump 142 is activated, the membrane cartridge 110 can be kept in a negative pressure state. Preferably, when the membrane cartridge coupler 120 is coupled with the membrane cartridge 110, the inlet of the first needle tube 141 is located at the bottom of the membrane cartridge 110 and keeps a gap of 0-1 mm with the proximal end surface of the filter membrane 130, so that when the membrane cartridge 110 is placed in the sample liquid, the inlet of the first needle tube 141 can be in contact with the liquid surface of the sample liquid. Exemplarily, the inlet of the first needle tube 141 can be in contact with the proximal end surface of the filter membrane 130, as shown. Figure 5 Exemplarily, the inlet of the first needle tube 141 can also keep a gap with the proximal end surface of the filter membrane 130. The gap is, for example, within 1 mm, such as 0.1 mm, 0.2 mm, 0.5 mm, 0.8 mm, etc.
[0033] In the negative pressure filtration mode in the related art, the depth of the membrane cartridge 110 is large, and the extension length of the needle tube is short, so that when the membrane cartridge 110 is inserted into the sample cup 210, the inlet of the needle tube is far from the height of the sample liquid. After the negative pressure pump 142 is activated, only the gas in the membrane cartridge 110 can be extracted, and therefore a subsequent step of turning over the membrane cartridge 110 and extracting the sample liquid is required. In the sampling mechanism of the present embodiment, the first needle tube 141 is extended, and when the membrane cartridge coupler 120 is coupled with the membrane cartridge 110, the inlet of the first needle tube 141 is located at the bottom of the membrane cartridge 110 and keeps a gap of 0-1 mm with the proximal end surface of the filter membrane 130. In this way, when the membrane cartridge 110 is inserted into the sample cup 210, the air in the membrane cartridge 110 and the sample liquid immersed in the membrane cartridge 110 can be simultaneously extracted through the negative pressure pump 142 and the first needle tube 141. Since the target cells cannot pass through the filter membrane 130, they are adsorbed to the distal end surface of the filter membrane 130. Further, the inlet of the first needle tube 141 is located at the bottom of the membrane cartridge 110 and keeps a gap of 0-1 mm with the proximal end surface of the filter membrane 130, which can minimize the dead volume of the sample liquid and significantly reduce the residues of blood cells and mucus.
[0034] In some embodiments, the pressure control assembly further comprises a second needle tube 143 and a positive pressure pump 144, as shown. Figure 2 and Figure 5 The second needle tube 143 is in communication with the positive pressure pump 144. Exemplarily, the second needle tube 143 is in communication with the positive pressure pump 144 through a hose or a hard tube. When the positive pressure pump 144 is activated, the membrane cartridge 110 can be kept in a positive pressure state. Preferably, the second needle tube 143 and the first needle tube 141 are independent of each other. Since the first needle tube 141 also extracts the cell processing liquid, the independent arrangement of the second needle tube 143 and the first needle tube 141 can avoid the contamination of the second needle tube 143. When different types of cell samples are processed, only the first needle tube 141 needs to be cleaned or replaced, which can simplify the subsequent cleaning process.
[0035] The positive pressure filtration method in the related art can only process about 2 ml of sample liquid. For a sample with a low target cell content, it is difficult to enrich enough target cells for detection, which affects the reliability of the detection result. The sampling mechanism in the embodiment can continuously extract sample liquid through negative pressure, so that the amount of sample that can be processed is greatly increased. For a sample with a low target cell content, by increasing the sample amount, enough target cells can be enriched for detection, solving the diagnostic problem of a low cell amount sample.
[0036] On the other hand, in the related art, the positive pressure filtration method injects a buffer to resuspend the target cells on the surface of the filter membrane 130. In this process, part of the target cells remain on the surface of the filter membrane and cannot be completely transferred to the slide, which exacerbates the difficulty of enrichment of a low cell amount sample. The sampling mechanism in the embodiment can make the target cells on the lower surface of the filter membrane 130 fall off by starting the positive pressure pump 144 to introduce gas into the membrane cartridge 110, which is under positive pressure. This method can achieve the full amount of target cells on the filter membrane 130 to fall off, and for the preparation of a low cell amount sample, the sampling mechanism in the embodiment can reduce the risk of loss of target cells, thereby reducing the difficulty of enrichment of a low cell amount sample.
[0037] In some embodiments, the membrane cartridge coupler 120 further integrates a liquid injection assembly. When the membrane cartridge coupler 120 is coupled with the membrane cartridge 110, the liquid injection assembly is used to inject liquid into the membrane cartridge 110. For example, the liquid injection assembly is used to inject a buffer into the membrane cartridge 110. Since the lower surface of the filter membrane 130 adsorbs target cells, the filter holes on the filter membrane 130 are blocked. After the buffer is injected into the membrane cartridge 110, the buffer can form a liquid layer on the upper surface of the filter membrane 130. Then, the positive pressure pump 144 is started, and the buffer can pass through the filter membrane 130 under the action of pressure and carry the fallen target cells into the sedimentation bin 310, and then be transferred to the slide 320.
[0038] In some embodiments, the liquid injection assembly includes a third needle tube 151 and a plunger pump 152, and the third needle tube 151 and the plunger pump 152 are in communication, as shown in Figure 2 and Figure 5 For example, the plunger pump 152 is also connected with a buffer reagent barrel, and the buffer in the buffer reagent barrel can be pumped into the third needle tube 151 by the plunger pump 152, and then into the membrane cartridge 110. Preferably, the third needle tube 151 is independent of the first needle tube 141 and the second needle tube 143, as shown in Figure 5 In this way, the risk of cross contamination of the first needle tube 141, the second needle tube 143 and the third needle tube 151 can be avoided.
[0039] The sampling mechanism of the embodiment, in the wafer preparation stage, combines the liquid layer and the positive pressure technology, and the target cells on the filter membrane 130 are transferred to the settling bin 310 by the positive pressure flushing of the buffer. In this process, on the one hand, the target cells and the gland cell groups can be prevented from deforming in the transfer process, so as to protect the cell integrity; on the other hand, the full transfer of the target cells on the filter membrane 130 can be realized; and thirdly, the uniformity of the target cell settling can be improved. Specifically, the existing wafer preparation process adopts the pipette fixed-point sampling method, which is easy to cause the phenomenon that the cells in the middle region of the glass slide 320 are overlapped, and the cells in the peripheral region are sparse. The embodiment realizes the uniform transfer of the target cells to the settling bin 310 in a surface form by combining the surface cell distribution of the filter membrane 130 with the positive pressure flushing technology, so that the target cell suspension in the settling bin 310 can present a uniform spatial distribution state, and finally a single-layer cell coating with uniform target cell density and no regional aggregation is formed on the glass slide 320, thereby providing a high-quality wafer sample for subsequent pathological diagnosis.
[0040] In some embodiments, the outlets of the second needle tube 143 and the third needle tube 151 are higher than the inlet of the first needle tube 141, as shown in FIG. 2. When the membrane cartridge 110 is immersed in the sample liquid, the outlets of the second needle tube 143 and the third needle tube 151 are located above the sample liquid, so that the second needle tube 143 and the third needle tube 151 are not in contact with the sample, thereby avoiding cross contamination by physical isolation. For example, as shown in FIG. 3, the outlet of the second needle tube 143 is close to the surface of the membrane cartridge coupler 120, and the third needle tube 151 protrudes from the membrane cartridge coupler 120. The length of the third needle tube 151 is between the first needle tube 141 and the second needle tube 143, which not only can be isolated from the sample, but also is beneficial to the formation of a uniform liquid layer on the surface of the filter membrane 130. Figure 5 Figure 5
[0041] In some embodiments, the membrane cartridge coupler 120 further integrates a pressure detection assembly 160, as shown in FIG. 4. The pressure detection assembly 160 includes a pressure sensor. The pressure sensor is arranged in the second needle tube 143 of the pressure control assembly. When the membrane cartridge coupler 120 is coupled with the membrane cartridge 110, the pressure detection assembly 160 is used to detect the pressure in the membrane cartridge 110. Figure 2
[0042] When the target cells are adsorbed on the filter membrane 130, the pressure detection assembly 160 can sense the pressure in the membrane cylinder 110, and by precisely controlling the duration of negative pressure, it can ensure that a sufficient amount of target cells is captured; for samples with a small amount of target cells, the negative pressure suction time can be continuously adjusted to improve the cell capture efficiency and basically achieve the capture of all target cells. For example, when the pressure in the membrane cylinder 110 is -5kPa~ -60kPa, it is maintained for 0.5~30s to complete the adsorption of target cells. The sampling mechanism of the present embodiment, through the setting of the pressure detection assembly 160, not only realizes the stable and controllable quantity of the settled target cells, but also optimizes the repeatability of the cell enrichment process through a closed-loop feedback mechanism.
[0043] On the other hand, the sampling mechanism of the present embodiment, through the setting of the pressure detection assembly 160, can also monitor whether the membrane cylinder 110 leaks, whether the membrane cylinder 110 is successfully coupled, and whether the filter membrane 130 is damaged. For example, when the surface of the filter membrane 130 is adsorbed and completely covered by target cells, the negative pressure will rise rapidly, and by the standard curve of the negative pressure value and the target cell value, the approximate number of adsorbed cells can be determined according to the negative pressure rising speed and the negative pressure value. For example, the pressure detection assembly 160 monitors the pressure change in the membrane cylinder 110 in real time, and when the pressure curve deviates from the preset threshold, the membrane cylinder coupler 120 automatically decouples and disposes the abnormal membrane cylinder 110, and then starts the coupling process of the new membrane cylinder 110. For example, the negative pressure comparison method before and after taking the membrane cylinder 110 is used to accurately verify whether the membrane cylinder coupler 120 and the membrane cylinder 110 are successfully coupled, if not, continue to automatically couple, if not successfully coupled for several times, intelligently trigger an alarm, and at the same time generate a traceable process parameter log (including fields such as time stamp, pressure value, coupling times, etc.), providing a data basis for intelligent manufacturing.
[0044] In some embodiments, the membrane cylinder 110 includes a first cylinder body 112 and a second cylinder body 113 connected to each other, as shown in Figure 6 For example, the first cylinder body 112 and the second cylinder body 113 are integrally formed by injection molding. The first cylinder body 112 is used to fix the filter membrane 130, and the second cylinder body 113 is used to couple with the membrane cylinder coupler 120. Preferably, the diameter of the first cylinder body 112 is smaller than the diameter of the second cylinder body 113, as shown in Figure 6 Because the diameter of the first cylinder body 112 is smaller, the spacing between adjacent two first cylinder bodies 112 is larger, so that not only it is easy to place multiple membrane cylinders 110 on the membrane cylinder support 180 at the same time, but also it is beneficial to keep the size of the membrane cylinder support 180 small.
[0045] In some embodiments, in addition to being used to install the membrane cylinder 110, the membrane cylinder support 180 is also provided with a membrane cylinder scraper 181, as shown in Figure 7As shown. The membrane tube scraper 181 has a structure with an opening on one side. When the membrane tube 110 is coupled with the membrane tube coupler 120, the first tube body 112 or the second tube body 113 of the membrane tube 110 is inserted into the membrane tube scraper 181, and the membrane tube 110 is fixed by the membrane tube scraper 181. Then the membrane tube coupler 120 is moved, thereby decoupling the membrane tube 110 from the membrane tube coupler 120.
[0046] In some embodiments, the membrane tube holder 180 also includes a cleaning tank 182. The cleaning tank 182 is used to clean the membrane tube coupler 120, and particularly to clean the first needle 141 on the membrane tube coupler 120. After the membrane tube 110 is decoupled from the membrane tube coupler 120, the membrane tube coupler 120 can be placed in the cleaning tank 182 for cleaning before being used for the next sample processing, thus avoiding cross-contamination between samples.
[0047] In some embodiments, the inner diameter of the second cylinder 113 matches the outer diameter of the membrane cylinder coupler 120, and when the membrane cylinder coupler 120 is inserted into the membrane cylinder 110, the membrane cylinder coupler 120 is coupled to the membrane cylinder 110, such as... Figure 5 As shown. For example, the outer surface of the membrane coupler 120 and / or the inner surface of the second cylinder 113 are provided with an anti-slip coating, which prevents the membrane cylinder 110 from detaching when the membrane coupler 120 is coupled to the membrane cylinder 110. In this embodiment, the membrane coupler 120 and the membrane cylinder 110 are coupled by an interference fit, eliminating the need for other coupling structures and simplifying the structure of the membrane coupler 120 and the membrane cylinder 110.
[0048] In some embodiments, the membrane tube 110 further includes a connecting portion 114 for connecting the first tube 112 and the second tube 113, and the connecting portion 114 has an inclined structure, such as... Figure 5 and Figure 6 As shown. For example, the angle between the connecting portion 114 and the wall surface of the first cylinder 112 or the second cylinder 113 is 90° to 175°. For example, the angle between the connecting portion 114 and the wall surface of the first cylinder 112 or the second cylinder 113 is 120° to 150°. When transferring the membrane cylinder 110 from the packaging box to the membrane cylinder support 180, the inclined connecting portion 114 can act as a guide, making it easy to place the membrane cylinder 110 smoothly onto the membrane cylinder support 180. A schematic diagram of the membrane cylinder 110 placed on the membrane cylinder support 180 is shown below. Figure 7 As shown.
[0049] In some embodiments, the end of the membrane cartridge 110 coupled with the membrane cartridge coupler 120 is provided with a protrusion 111 protruding from the outer surface of the membrane cartridge 110 and increasing the width of the end surface of the membrane cartridge 110 to be greater than the thickness of the membrane cartridge 110. The membrane cartridge coupler 120 is formed with a stepped surface 121, and when the membrane cartridge 110 is coupled with the membrane cartridge coupler 120, the stepped surface 121 is in contact with the end surface of the membrane cartridge 110, as shown in Figure 5 and Figure 6 Preferably, when the membrane cartridge coupler 120 enters the membrane cartridge 110, the stepped surface 121 is in linear contact with the end surface of the protrusion 111. When the membrane cartridge coupler 120 continues to enter the membrane cartridge 110, the membrane cartridge coupler 120 presses the membrane cartridge 110 and causes plastic deformation of the membrane cartridge 110, so that the inner wall of the membrane cartridge 110 is tightly attached to the outer surface of the membrane cartridge coupler 120 to maintain the seal between the membrane cartridge coupler 120 and the membrane cartridge 110. At the same time, the stepped surface 121 is in contact with the surface of the protrusion 111, thereby facilitating further enhancing the seal between the membrane cartridge coupler 120 and the membrane cartridge 110 to facilitate controlling the negative pressure or positive pressure state in the membrane cartridge 110.
[0050] On the other hand, when the membrane cartridge 110 is coupled with the membrane cartridge coupler 120, the stepped surface 121 is in contact with the end surface of the membrane cartridge 110, which can limit the membrane cartridge coupler 120 from being inserted into the membrane cartridge 110 too much, thereby preventing the first needle tube 141 from puncturing the filter membrane 130.
[0051] The cell enrichment device of the present embodiment is used to complete the enrichment of cells and to deposit the enriched cells on a glass slide. As shown in Figure 1 , the cell enrichment device of the present embodiment includes a sampling mechanism 100, a sample mechanism 200, and a deposition mechanism 300, and through the cooperation of the three mechanisms, the enrichment of cells can be automatically completed and the enriched cells can be deposited on a glass slide.
[0052] In some embodiments, the sampling mechanism 100 is the sampling mechanism of any one of the technical solutions of the present embodiment.
[0053] Without limitation, the sampling mechanism 100 of the present embodiment further includes a first mechanical arm 170, as shown in Figure 1 and Figure 4 The membrane cartridge coupler 120 of the sampling mechanism 100 is installed below the first mechanical arm 170, and the movement of the first mechanical arm 170 can drive the membrane cartridge coupler 120 to move. For example, the first mechanical arm 170 can move in three directions XYZ in a three-dimensional space. The mechanism of the first mechanical arm 170 can be the same as the prior art, which will not be described here. The sampling mechanism 100 of the present embodiment further includes a membrane cartridge support 180, which is provided with a plurality of hole positions, and the membrane cartridge 110 is installed in the hole positions of the membrane cartridge support 180, as shown in Figure 1 and Figure 7 The membrane cartridge support 180 is provided with a plurality of hole positions, and the membrane cartridge 110 is installed in the hole positions of the membrane cartridge support 180.
[0054] In some embodiments, the sample mechanism 200 is used to hold the sample cup 210, such as Figure 1 and Figure 8 As shown. In the pretreatment stage, the sampling mechanism 100 moves to the sample cup 210, the membrane tube 110 is immersed in the sample cup 210, the sample liquid in the sample cup 210 is aspirated to maintain negative pressure in the membrane tube 110, and the target cells are adsorbed onto the filter membrane 130.
[0055] For example, the sample mechanism 200 includes a sample conveyor belt 220, such as Figure 1 and Figure 8 As shown. The sample cup 210 is placed on the sample conveyor belt 220, and the sample cup 210 can be transported to a predetermined station by the movement of the sample conveyor belt 220. The sample mechanism 200 also includes a cup-grabbing mechanical gripper 230 and a capping assembly 240, as shown. Figure 1 and Figure 3 As shown. The sample cup mechanical gripper 230 is used to remove the sample cup 210 from the sample conveyor belt 220 and place it in the capping station. The capping assembly 240 is used to unscrew the cap 211 of the sample cup 210 to expose the solution in the sample cup 210, so that the membrane cartridge 110 can be immersed in the sample cup 210 for sample pretreatment. Exemplarily, the capping assembly 240 includes a rotating mechanical gripper 241 for fixing the sample cup 210, such as... Figure 3 As shown. The structures of the sample cup 210, sample conveyor belt 220, cup-picking mechanical claw 230, and capping assembly 240 are the same as those in the prior art, and will not be described in detail here.
[0056] In some embodiments, the settling mechanism 300 includes a settling chamber 310 and a glass slide 320, such as Figure 1 and Figure 9 As shown. The structure of the settling chamber 310 and the glass slide 320 can be the same as in the prior art, and will not be described in detail here. The glass slide 320 is located at the bottom of the settling chamber 310, as shown. Figures 9 to 11 As shown. During the slide preparation stage, the sampling mechanism 100 moves to the settling chamber 310, and positive pressure is maintained inside the membrane cartridge 110 by introducing gas. The target cells detach from the filter membrane 130 and settle onto the glass slide 320. The settling chamber 310 has a hollow internal structure to allow the membrane cartridge 110 to be placed inside for settling, as shown. Figures 10 to 12 As shown.
[0057] Not limited to this, the settlement mechanism 300 also includes a settlement plate 330, such as Figure 9 and Figure 10The structure of the sedimentation disc 330 can be the same as the prior art, and will not be described here. The sedimentation disc 330 is distributed with a plurality of sedimentation chambers 310 in the circumferential direction, and the bottom of each sedimentation chamber 310 has a glass slide 320, so that the sedimentation mechanism 300 of the present application can be used for the sedimentation of multiple samples at the same time, as shown in Figures 9 to 11
[0058] The cell enrichment device of the present embodiment has the sampling mechanism of any one of the technical solutions in the present embodiment. In the pretreatment stage, the membrane cylinder 110 maintains negative pressure, which can adsorb the target cells to the distal end surface of the filter membrane 130, and at the same time, the sample liquid in the sample cup 210 can be pumped out. Compared with the prior art, not only the step of turning over the membrane cylinder 110 to pump out the sample liquid can be omitted, but also the structure of the cell enrichment device can be simplified. In addition, the sample liquid can be continuously pumped out by negative pressure, so that the amount of samples that can be processed is greatly increased. For samples with low content of target cells, by increasing the sample amount, enough target cells can be enriched for detection, improving the efficiency and reliability of cell pretreatment, and solving the diagnosis problem of low cell amount samples.
[0059] On the other hand, in the tablet preparation stage, the membrane cylinder 110 maintains positive pressure, which can make the target cells adsorbed on the filter membrane 130 fall off. This method can realize the full amount of target cells on the filter membrane 130 to fall off, and can reduce the loss of target cells for low cell amount sample preparation, thereby further reducing the difficulty of enrichment of low cell amount samples.
[0060] The cell enrichment method of the present embodiment is realized by using the cell enrichment device of any one of the technical solutions in the present embodiment. The cell enrichment method of the present embodiment includes the following steps: Step 100: The cup cover 211 of the sample cup 210 is unscrewed, the membrane cylinder coupler 120 of the sampling mechanism 100 is coupled with the membrane cylinder 110, and then the membrane cylinder 110 is transferred to the sample cup 210 with the cup cover 211 opened.
[0061] Step 200: The membrane cylinder 110 is lowered and the filter membrane 130 is immersed in the sample liquid. The negative pressure pump 142 is started to maintain negative pressure in the membrane cylinder 110, and the target cells are adsorbed on the filter membrane 130. For example, the pressure in the membrane cylinder 110 is-5kPa~-60kPa. Specifically, taking the standard atmospheric pressure 101kPa as zero point, the pressure in the membrane cylinder 110 is 5kPa~60kPa lower than the standard atmospheric pressure. For example, the pressure in the membrane cylinder 110 is-10kPa~-30kPa.
[0062] Step 300: After aspiration is complete, the membrane cartridge 110 is transferred to the settling chamber 310. After turning off the negative pressure pump 142, the positive pressure pump 144 is activated to maintain positive pressure inside the membrane cartridge 110. The target cells detach from the filter membrane 130 and settle onto the glass slide 320. For example, the pressure inside the membrane cartridge 110 is 5 kPa to 60 kPa. Specifically, with standard atmospheric pressure of 101 kPa as zero, the pressure inside the membrane cartridge 110 is 5 kPa to 60 kPa higher than standard atmospheric pressure. For example, the pressure inside the membrane cartridge 110 is 10 kPa to 30 kPa.
[0063] The cell enrichment method in this embodiment not only eliminates the step of flipping the membrane tube to aspirate the sample solution, but also simplifies the structure of the cell enrichment device; it also reduces the difficulty of enriching samples with low cell counts.
[0064] In some embodiments, after immersing the filter membrane 130 in the sample solution in step 200 and before starting the negative pressure pump 142, the following steps are also included: starting the positive pressure pump 144, injecting gas into the sample solution in the sample cup 210 through the second needle tube 143, and maintaining positive pressure inside the membrane cylinder 110. Before starting the negative pressure pump 142, introducing gas into the sample cup 210 can agitate the sample solution in the sample cup 210, causing cells settled at the bottom of the sample cup 210 to float, thereby increasing the cell collection volume on the filter membrane 130.
[0065] For example, the pressure inside the membrane cartridge 110 is 5 kPa to 60 kPa, and the positive pressure is maintained for 0.5 to 30 seconds. This not only allows the cells that have settled at the bottom of the sample cup 210 to float up, but also avoids the potential risk of sample liquid overflowing from the sample cup 210 due to excessive pressure.
[0066] In some embodiments, after aspiration, the following steps are further included: turning off the negative pressure pump 142, starting the plunger pump 152, and injecting buffer solution into the membrane cartridge 110. For example, 0.2~2 mL of buffer solution is injected into the membrane cartridge 110. The negative pressure pump 142 is then started again, and the buffer solution in the membrane cartridge 110 is aspirated through the first syringe 141. This process washes the target cells. Specifically, during the pretreatment stage, some blood cells are adsorbed onto the filter membrane 130. By injecting buffer solution into the membrane cartridge 110, the blood cells can be resuspended in the buffer solution; then, by starting the negative pressure pump 142, the blood cells are discharged from the first syringe 141 along with the buffer solution, resulting in higher purity of the target cells attached to the filter membrane 130, and thus a cleaner and clearer background of the target cells in the resulting slide, which improves the detection effect.
[0067] In some embodiments, after the negative pressure pump 142 is closed and before the positive pressure pump 144 is started, the method further comprises the following steps: starting the plunger pump 152, injecting the buffer into the membrane cartridge 110, and forming a buffer layer on the filter membrane 130. For example, 0.2-2 mL of buffer is injected into the membrane cartridge 110, and after the buffer layer is formed on the surface of the filter membrane 130, the positive pressure pump 144 is started.
[0068] In the cell enrichment method of the present embodiment, the target cells on the filter membrane 130 are completely transferred to the sedimentation bin 310 by the buffer positive pressure flushing. In this process, on the one hand, the target cells and the gland cell mass can be prevented from deforming during the transfer process, thereby protecting the integrity of the cells; on the other hand, the full amount of the target cells on the filter membrane 130 can be transferred; and thirdly, the uniformity of the target cell sedimentation can be improved.
[0069] In some embodiments, after the negative pressure pump 142 is started, the number of adsorbed target cells is determined based on the pressure value detected by the pressure detection assembly 160. When the surface of the filter membrane 130 is covered by the target cells, the pressure in the membrane cartridge 110 can rapidly decrease. For example, when the pressure in the membrane cartridge 110 rapidly decreases from -0.1 kPa to -20 kPa, it can be considered that the surface of the filter membrane 130 is completely covered by the target cells. Further, a standard curve of the pressure in the membrane cartridge 110 and the number of target cells can be prepared, and based on the standard curve, the number of target cells adsorbed on the surface of the filter membrane 130 can be determined according to the pressure in the membrane cartridge 110.
[0070] In some embodiments, before the first needle tube 141 is used for sample processing, the first needle tube 141 is cleaned by using one or more of an acidic cleaning solution, an alkaline cleaning solution, and ultrasonic cleaning. The acidic cleaning solution and the alkaline cleaning solution can be the same as those in the prior art, and the process of ultrasonic cleaning can be the same as that in the prior art, which will not be described here. By cleaning the first needle tube 141, cross-infection between different samples can be avoided.
[0071] The above merely illustrates the specific embodiments of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art can easily think of changes or replacements within the technical scope disclosed by the present application, which shall be covered within the protection scope of the present application.
Claims
1. A sampling mechanism for use in a cell enrichment device, characterized in that, The sampling mechanism includes a membrane tube (110), a membrane tube coupler (120), and a filter membrane (130). The membrane tube (110) has an open structure at both ends. The filter membrane (130) is fixed at one end of the membrane tube (110). The membrane tube coupler (120) can be coupled or decoupled from the other end of the membrane tube (110). The membrane coupler (120) is equipped with a pressure control component. When the membrane coupler (120) is coupled to the membrane tube (110), in the pretreatment stage, the pressure control component is used to maintain a negative pressure inside the membrane tube (110) and simultaneously extract the sample liquid. In the slide preparation stage, the pressure control component is used to maintain a positive pressure inside the membrane tube (110).
2. The sampling mechanism according to claim 1, characterized in that, The membrane coupler (120) is also integrated with a liquid injection assembly. When the membrane coupler (120) is coupled to the membrane (110), the liquid injection assembly is used to inject liquid into the membrane (110).
3. The sampling mechanism according to claim 2, characterized in that, The pressure control assembly includes a first needle tube (141) and a negative pressure pump (142). The first needle tube (141) is connected to the negative pressure pump (142). When the membrane coupler (120) is coupled to the membrane tube (110), the inlet of the first needle tube (141) is located at the bottom of the membrane tube (110) and maintains a gap of 0~1mm between it and the proximal end face of the filter membrane (130).
4. The sampling mechanism according to claim 3, characterized in that, The pressure control assembly also includes a second needle tube (143) and a positive pressure pump (144), the second needle tube (143) being connected to the positive pressure pump (144) and the second needle tube (143) being independent of the first needle tube (141).
5. The sampling mechanism according to claim 4, characterized in that, The injection assembly includes a third needle tube (151) and a plunger pump (152), the third needle tube (151) and the plunger pump (152) being connected, the third needle tube (151) being used to inject buffer solution into the membrane tube (110), and the third needle tube (151) being independent of the first needle tube (141) and the second needle tube (143).
6. The sampling mechanism according to claim 5, characterized in that, The outlets of the second needle (143) and the third needle (151) are higher than the inlet of the first needle (141), and when the membrane tube (110) is immersed in the sample liquid, a gap is maintained between the outlets of the second needle (143) and the third needle (151) and the liquid surface of the sample liquid.
7. The sampling mechanism according to claim 1, characterized in that, The membrane coupler (120) also integrates a pressure detection component (160), which includes a pressure sensor located inside the second needle tube (143) of the pressure control component. When the membrane coupler (120) is coupled to the membrane (110), the pressure detection component (160) is used to detect the pressure inside the membrane (110).
8. The sampling mechanism according to claim 1, characterized in that, The membrane tube (110) includes a first tube (112) and a second tube (113) connected to each other. The first tube (112) is used to fix the filter membrane (130), and the second tube (113) is used to couple with the membrane tube coupler (120). The diameter of the first tube (112) is smaller than the diameter of the second tube (113).
9. The sampling mechanism according to claim 8, characterized in that, The membrane tube (110) further includes a connecting part (114), which is used to connect the first tube (112) and the second tube (113), and the connecting part (114) has an inclined structure.
10. The sampling mechanism according to claim 8, characterized in that, The inner diameter of the second cylinder (113) matches the outer diameter of the membrane cylinder coupler (120), and when the membrane cylinder coupler (120) is inserted into the membrane cylinder (110), the membrane cylinder coupler (120) is coupled to the membrane cylinder (110).
11. The sampling mechanism according to claim 10, characterized in that, The membrane tube (110) is coupled to the membrane tube coupler (120) at one end with a protruding edge (111). The protruding edge (111) protrudes from the outer surface of the membrane tube (110) and makes the width at the end face of the membrane tube (110) greater than the thickness of the membrane tube (110). A stepped surface (121) is formed on the membrane tube coupler (120). When the membrane tube (110) is coupled to the membrane tube coupler (120), the stepped surface (121) contacts the end face of the membrane tube (110).
12. A cell enrichment device, characterized in that, It includes a sampling mechanism (100), a sample collection mechanism (200), and a settling mechanism (300), wherein, The sampling mechanism (100) is the sampling mechanism according to any one of claims 1 to 11; The sample mechanism (200) is used to place the sample cup (210). In the pretreatment stage, the sampling mechanism (100) moves to the sample cup (210), the membrane tube (110) is immersed in the sample cup (210), and the sample liquid in the sample cup (210) is aspirated to maintain negative pressure in the membrane tube (110) and cause the target cells to be adsorbed onto the filter membrane (130). The sedimentation mechanism (300) includes a sedimentation chamber (310) and a glass slide (320). The glass slide (320) is located at the bottom of the sedimentation chamber (310). During the slide preparation stage, the sampling mechanism (100) moves to the sedimentation chamber (310) and maintains positive pressure inside the membrane tube (110) by introducing gas. The target cells detach from the filter membrane (130) and settle onto the glass slide (320).
13. A method for cell enrichment using the cell enrichment device of claim 12, characterized in that, Includes the following steps: Unscrew the lid (211) of the sample cup (210), and after the membrane tube coupler (120) of the sampling mechanism (100) is coupled to the membrane tube (110), the membrane tube (110) is transferred to the sample cup (210) with the lid already opened. The membrane tube (110) is lowered, and the filter membrane (130) is immersed in the sample solution. The negative pressure pump (142) is started to maintain negative pressure inside the membrane tube (110) and cause the target cells to be adsorbed onto the filter membrane (130). After the aspiration is completed, the membrane tube (110) is transferred to the sedimentation chamber (310). After the negative pressure pump (142) is turned off, the positive pressure pump (144) is started to keep the membrane tube (110) under positive pressure. The target cells fall off the filter membrane (130) and settle on the glass slide (320).
14. The method for cell enrichment according to claim 13, characterized in that, Before starting the negative pressure pump (142), the following steps are also included: starting the positive pressure pump (144), injecting gas into the sample liquid in the sample cup (210) through the second needle tube (143), and maintaining positive pressure in the membrane tube (110).
15. The method for cell enrichment according to claim 13, characterized in that, After aspiration is completed, the following steps are also included: turn off the negative pressure pump (142), start the plunger pump (152), and inject buffer solution into the membrane cartridge (110); start the negative pressure pump (142) again and draw the buffer solution in the membrane cartridge (110) through the first needle (141).
16. The method for cell enrichment according to any one of claims 13 to 15, characterized in that, After the negative pressure pump (142) is turned off and before the positive pressure pump (144) is started, the following steps are also included: starting the plunger pump (152) to inject buffer solution into the membrane cartridge (110) and forming a buffer solution layer on the filter membrane (130).
17. The method for cell enrichment according to any one of claims 13 to 15, characterized in that, After the negative pressure pump (142) is started, the number of target cells adsorbed is determined based on the pressure value detected in the membrane tube (110) by the pressure detection component (160). And / or, after starting the negative pressure pump (142), maintain the pressure inside the membrane cylinder (110) at -5kPa to -60kPa; And / or, after starting the positive pressure pump (144), maintain the pressure inside the membrane cylinder (110) at 5 kPa to 60 kPa; And / or, start the plunger pump (152) to inject 0.2~2 mL of buffer solution into the membrane cartridge (110).
18. The method for cell enrichment according to any one of claims 13 to 15, characterized in that, Before the first syringe (141) is used for the next sample processing, the following steps are also included: cleaning the first syringe (141) with one or more of acidic cleaning solution, alkaline cleaning solution, and ultrasonic cleaning.