Online cell counting device and control method

Abstract

The invention relates to the technical field of cell counting, and provides an online cell counting device and a control method.The online cell counting device comprises a device body, a detection assembly, a fluid driving assembly and a connecting pipeline, and the detection assembly and the fluid driving assembly are installed on the device body; the detection assembly is communicated with the fluid driving assembly through the connecting pipeline, and the fluid driving assembly is communicated with a to-be-detected container through the connecting pipeline and enables to-be-detected cells in the to-be-detected container to enter the detection assembly. Cells in the to-be-detected container are directly conveyed into the detection assembly to be detected through the fluid driving assembly, online real-time monitoring is achieved, the manual sampling process is avoided, the sterile environment is guaranteed, and meanwhile accurate detection and cell recycling of high-concentration cells can be achieved through cooperation of the connecting pipeline and the fluid driving assembly.

Description

Technical Field

[0001] The present invention relates to the technical field of cell counting, in particular to an online cell counting device. Background Art

[0002] Most existing cell counters, such as the Thermo Countess 3, are offline. These offline cell counters require manual removal of cells, placement of a detection chip, and then placement of the chip into the instrument for imaging and identification. This method presents the following challenges: First, the cell removal process is time-sensitive and cannot reflect the cell concentration and status during the encapsulation process in real time. Second, the cells are exposed to the external environment after removal, posing a risk of contamination and failing to maintain a sterile environment.

[0003] In addition, offline cell counters cannot effectively and accurately detect high-concentration cells (concentration greater than 5E7 / ml). This is because high-concentration cells tend to overlap on the detection chip, resulting in omissions during photo recognition. To solve this problem, the cell solution usually needs to be diluted before testing, which increases the number of operation steps and time costs.

[0004] Another issue worth noting is that existing offline cell counting methods cannot effectively recover some rare cells, which not only wastes precious samples but also limits the research and application of these cells.

[0005] Existing cell counting technologies also have several limitations. For example, they lack automation and real-time monitoring capabilities, making them incapable of meeting the online monitoring needs of continuous production processes. Furthermore, existing technologies lack flexibility and adaptability when processing cells of varying types and sizes, making them difficult to meet diverse research and production needs.

[0006] Furthermore, existing cell counting equipment is often bulky and difficult to integrate into other systems, limiting its application in complex experimental or production environments. Furthermore, the operation of these devices often requires professional training, increasing the cost and barrier to entry.

[0007] In view of the above problems, an online cell counting device is proposed. Summary of the Invention

[0008] The purpose of the present invention is to provide an online cell counting device with the advantages of real-time monitoring of cell concentration and status, ensuring a sterile environment, accurately detecting high-concentration cells, realizing cell recycling, improving the degree of automation, adapting to multiple cell types, being easy to integrate and simple to operate.

[0009] To achieve the above object, the present invention adopts the following technical solutions:

[0010] An online cell counting device according to an embodiment of the present invention includes: a fluid circuit unit comprising a microfluidic chip, a fluid drive assembly, and a connecting pipeline, the connecting pipeline being connected to the microfluidic chip, the fluid drive assembly being disposed on the connecting pipeline and configured to drive fluid to flow in the microfluidic chip; an optical circuit unit comprising a laser light source and a focusing assembly, wherein emitted light from the laser light source is focused by the focusing assembly into a light plate and then incident on the microfluidic chip; a detection assembly disposed on a side of the microfluidic chip facing away from the focusing assembly; a control unit configured to control the operating states of the fluid drive assembly, the pinch valve on the connecting pipeline, and the detection assembly, and to obtain a cell counting result based on an electrical signal provided by the detection assembly; and the detection assembly comprising an XY adjustment mechanism and a photoelectric sensor mounted on the XY adjustment mechanism, the XY adjustment mechanism being configured to fine-tune the position of the photoelectric sensor along the X-axis and the Y-axis, respectively.

[0011] The online cell counting device according to an embodiment of the present invention integrates a fluidic unit, an optical unit, a detection component, and a control unit to form a closed-loop control system. It uses a microfluidic chip to automatically transport and detect cell samples, and incorporates an adjustable photoelectric sensor positioning mechanism to ensure detection accuracy. This device offers advantages such as enabling real-time online monitoring of the cell culture process, eliminating the risk of contamination introduced by manual operation, improving the detection accuracy of high-concentration cell samples, eliminating the need for sample dilution, supporting the recovery of precious samples, and enhancing system integration and automation.

[0012] In addition, the online cell counting device according to the above embodiment of the present invention may also have the following additional technical features:

[0013] In some embodiments of the present invention, the XY adjustment mechanism includes an aperture assembly, an X-axis adjustment member, a Y-axis adjustment member, a pressure block, a housing bracket, a bracket base, and a pressure cover; the aperture assembly includes an aperture, a lens, and a lens pressure cover, the lens has a lens bracket, the aperture and the lens pressure cover are both mounted on the lens bracket, and are respectively located on opposite sides of the lens; the bracket housing is fixed to the bracket base, the pressure block and the pressure cover are respectively arranged on opposite sides of the housing bracket, and are both fixedly connected to the housing bracket; the housing bracket is provided with a through hole, one end of the aperture assembly is inserted into the through hole of the housing bracket, the X-axis adjustment member and the Y-axis adjustment member are both mounted on the housing bracket, and are used to fine-tune the position of the aperture assembly along the X-axis direction and the Y-axis direction, respectively.

[0014] In some embodiments of the present invention, an X-axis linear module and a chip adjustment mechanism are also included; the chip adjustment mechanism is installed on the X-axis linear module and can move back and forth along the X-axis direction; the conical positioning pins of the chip adjustment mechanism cooperate with the positioning holes on the microfluidic chip to fine-tune the position of the microfluidic chip along the Z-axis direction.

[0015] In some embodiments of the present invention, the fluid drive assembly includes a first peristaltic pump and a second peristaltic pump, wherein the first peristaltic pump is used to pump the liquid in the container to be tested into the microfluidic chip; the second peristaltic pump is used to pump the liquid in the dilution container into the microfluidic chip.

[0016] In some embodiments of the present invention, the microfluidic chip has a cell detection channel and a first inlet end, a second inlet end and an outlet end connected to the cell detection channel; the connecting pipeline includes a first pipeline, a second pipeline and a third pipeline, one end of the first pipeline is connected to the container to be tested, the other end of the first pipeline is connected to one end of the third pipeline through a connector, the other end of the third pipeline is connected to the first inlet end of the microfluidic chip, one end of the second pipeline is connected to the dilution container, and the other end of the second pipeline is connected to the second inlet end of the microfluidic chip; the first peristaltic pump is arranged on the first pipeline, the second peristaltic pump is arranged on the second pipeline, and the pinch valve is arranged on the third pipeline.

[0017] In some embodiments of the present invention, a waste liquid storage device is further included, and the connecting pipeline further includes a fourth pipeline and a fifth pipeline, one end of each of the fourth pipeline and the fifth pipeline is connected to the waste liquid storage device, the other end of the fourth pipeline is connected to the outlet end of the microfluidic chip, and the other end of the fifth pipeline is connected to the first pipeline and the third pipeline through the connecting piece; the pinch valve is a two-way pinch valve, and the opening and closing of the third pipeline and the fifth pipeline are both controlled by the pinch valve.

[0018] In some embodiments of the present invention, a device body is further included, which includes a base plate, a bracket and a shell, the bracket is installed on the base plate, the detection component is installed on the bracket, the bottom surface of the shell is fixed to the base plate, and the fluid drive component and the switch component are both installed on the top surface of the shell.

[0019] In some embodiments of the present invention, the laser light source is mounted on the bracket, the X-axis linear module is mounted on the base plate below the bracket, a chip slot is provided on the bracket, the chip slot is located at the end of the focusing assembly away from the laser light source, the microfluidic chip is mounted in the chip slot, the liquid path unit is connected to the dilution container, the container to be tested and the waste liquid storage device, and the laser light source, the focusing assembly, the microfluidic chip, the aperture assembly and the photoelectric sensor are located on the same straight line.

[0020] In some embodiments of the present invention, the control unit includes a control circuit board and a display screen, the display screen is mounted on the top surface of the housing, the control circuit board is mounted on the bracket, and the control circuit board is electrically connected to the detection component, the moving component, the fluid drive component, the pinch valve and the display screen.

[0021] In some embodiments of the present invention, a heat dissipation assembly is further included, which includes a first heat dissipation member, a second heat dissipation member and a third heat dissipation member. The first heat dissipation member is installed on the laser light source, and the second heat dissipation member and the third heat dissipation member are both installed on the base plate, and the second heat dissipation member is located in the middle of the base plate, and the third heat dissipation member is located at the edge of the base plate.

[0022] The present invention also provides a method for controlling online cell counting, comprising:

[0023] Step S1: providing the above-mentioned online cell counting device;

[0024] Step S2: connecting the container to be tested, the dilution container, and the waste liquid storage device to the online cell counting device;

[0025] Step S3: Inserting the microfluidic chip:

[0026] Step S4: turning on the first peristaltic pump to allow the cell fluid to fill the connecting pipeline to rinse the connecting pipeline;

[0027] Step S5: selecting a counting mode according to the cell concentration and starting online cell counting detection;

[0028] Step S6: transporting the detected cells to the waste liquid storage device for collection and processing.

[0029] According to some embodiments of the present invention, the above-mentioned online cell counting control method can detect the number of cells in real time, and at the same time, the cells are directly transported to the detection chip, thereby avoiding the contamination of the cells by the external environment and the impact on the detection structure. At the same time, the accuracy of the detection results is improved by rinsing the connecting pipelines. The above-mentioned control method improves the convenience of users in detecting cells.

[0030] In some embodiments of the present invention, selecting a counting mode according to the cell concentration and starting online cell counting detection includes:

[0031] Step S51: detecting or inputting the cell concentration, and selecting a counting mode according to the cell concentration;

[0032] Step S52: If the low concentration counting mode is selected, controlling the fluid drive assembly to turn on the first peristaltic pump to transport the cells to be tested to the microfluidic chip;

[0033] If the high concentration counting mode is selected, the first peristaltic pump and the second peristaltic pump are turned on to deliver the cells to be tested to the microfluidic chip, and at the same time, a diluent is delivered into the microfluidic chip to adjust the concentration of the cells to be tested;

[0034] Step S53: turning on the laser light source to allow the laser to pass through the focusing assembly and enter the focusing detection channel of the microfluidic chip;

[0035] Step S54: adjusting the height of the microfluidic chip so that the light passing through the focused detection channel is incident on the sensing surface of the photoelectric sensor, thereby achieving cell counting.

[0036] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 Schematic diagram of the structure of an online cell counting device according to an embodiment of the present invention Figure 1 ;

[0038] Figure 2 Schematic diagram of the structure of an online cell counting device according to an embodiment of the present invention Figure 2 ;

[0039] Figure 3 Schematic diagram of the structure of an online cell counting device according to an embodiment of the present invention Figure 3 ;

[0040] Figure 4 Schematic diagram of the structure of an online cell counting device according to an embodiment of the present invention Figure 4 ;

[0041] Figure 5 Schematic diagram of the structure of the XY adjustment mechanism of an embodiment of the present invention Figure 1 ;

[0042] Figure 6 Schematic diagram of the structure of the XY adjustment mechanism of an embodiment of the present invention Figure 2 ;

[0043] Figure 7 This is a connection diagram of an online cell counting device according to an embodiment of the present invention;

[0044] Figure 8 The control method flow of the online cell counting device according to the embodiment of the present invention is as follows Figure 1 ;

[0045] Figure 9 The control method flow of the online cell counting device according to the embodiment of the present invention is as follows Figure 2 ;.

[0046] Reference numerals

[0047] 100. Online cell counting device; 1. Power socket; 2. Power switch; 3. Network socket; 4. Connector; 5. Microfluidic chip; 6. Fluid drive assembly; 7. Vent; 8. Laser light source; 9. Focusing assembly; 10. Pinch valve; 11. XY adjustment mechanism; 12. Photoelectric sensor; 13. Aperture assembly; 14. X-axis adjustment member; 15. Y-axis adjustment member; 16. Pressure block; 17. Housing bracket; 18. Bracket base; 19. Pressure cover; 20. Aperture; 21. USB port; 22. Lens pressure cover; 23. Lens bracket; 24. Through hole; 25. X-axis linear module; 26. Chip adjustment mechanism; 29. ​​First peristaltic pump; 30. Second peristaltic pump; 31. Cell detection flow channel; 32. First inlet port; 33. Second inlet port; 34. Outlet port; 35. First pipeline; 36. Second pipeline; 37. Third pipeline; 38. Container to be tested; 39. Dilution container; 40. Waste liquid storage device; 41. Fourth pipeline; 42. Fifth pipeline; 44. Bottom plate; 45. Bracket; 46. Housing; 47. Waste liquid box; 48. Control circuit board; 49. Display screen; 50. Connector; 51. First heat sink; 52. Second heat sink; 53. Third heat sink. DETAILED DESCRIPTION

[0048] The following describes an online cell counting device and online cell counting control method of the present invention in more detail with reference to the accompanying drawings. Preferred embodiments of the present invention are shown, and it should be understood that those skilled in the art may modify the present invention described herein while still achieving the advantageous effects of the present invention. Therefore, the following description should be understood as generally known to those skilled in the art and is not intended to limit the present invention.

[0049] In the description of this specification, "one embodiment" or "some embodiments" means that one or more embodiments of this specification include a particular feature, structure, or characteristic described in conjunction with the embodiment. Therefore, the phrases "in one embodiment," "in some embodiments," "in other embodiments," "in other embodiments," etc. that appear in different places in this specification do not necessarily refer to the same embodiment, but rather mean "one or more but not all embodiments," unless otherwise specifically emphasized.

[0050] The following describes embodiments of the present invention in detail. Examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended only to explain the present invention and are not to be construed as limiting the present invention.

[0051] In existing technologies, cell counters mostly use an offline detection mode, requiring manual transfer of samples to a detection chip for photographic identification. This method suffers from poor timeliness and a high risk of contamination. Especially for high-concentration cell samples, counting errors due to cell overlap are prone to occur, and sample recovery is impossible. Traditional equipment lacks automated control capabilities, making it difficult to meet the needs of real-time monitoring in a sterile environment. It also has limitations such as bulky size and complex operation.

[0052] To address these issues, researchers realized the need to build an integrated online detection system. By analyzing the key aspects of sample exposure during offline operations, they proposed embedding the detection process within a closed fluidic system. To address the challenges of high-concentration cell detection, they considered using fluid dynamic focusing technology to optimize cell distribution. Further research revealed that dynamically adjusting the positional relationship between the optical path and the detection element could improve signal acquisition accuracy, leading to the development of a technical concept combining fluidic control, optical detection, and automatic adjustment.

[0053] Therefore, the present invention proposes an online cell counting device 100 . The online cell counting device 100 according to an embodiment of the present invention will be described below with reference to the accompanying drawings.

[0054] According to the online cell counting device 100 of the embodiment of the present invention, Figure 1-Figure 4 As shown, it includes: a liquid circuit unit (not marked in the figure), including a microfluidic chip 5, a fluid driving component 6 and a connecting pipeline (not marked in the figure), the connecting pipeline is connected to the microfluidic chip 5, and the fluid driving component 6 is arranged on the connecting pipeline to drive the fluid to flow in the microfluidic chip 5;

[0055] An optical path unit (not marked in the figure) includes a laser light source 8 and a focusing component 9. The emitted light of the laser light source 8 is focused into a light plate by the focusing component 9 and then incident on the microfluidic chip 5.

[0056] A detection component (not marked in the figure) is provided on the side of the microfluidic chip 5 facing away from the focusing component 9;

[0057] a control unit (not shown), configured to control the working states of the fluid drive assembly 6, the pinch valve 10 on the connecting pipeline, and the detection assembly, and to obtain a cell counting result based on an electrical signal provided by the detection assembly;

[0058] The detection assembly includes an XY adjustment mechanism 11 and a photoelectric sensor 12 mounted on the XY adjustment mechanism 11 . The XY adjustment mechanism 11 is used to fine-tune the position of the photoelectric sensor 12 along the X-axis direction and the Y-axis direction respectively.

[0059] Among them, the microfluidic chip 5 refers to a transparent substrate device with a micron-level flow channel, which can be made of polydimethylsiloxane material through a soft lithography process and is used to constrain the flow of cell monolayers. The fluid drive component 6 refers to a device that generates fluid power, which can be implemented by a peristaltic pump or a syringe pump to ensure continuous and stable delivery of samples. The light plate refers to a sheet of light formed by a cylindrical lens, which can be specifically implemented by a semiconductor laser with a wavelength of 635nm in combination with an aspheric lens group, and is used to illuminate the flow cells laterally. The XY adjustment mechanism 11 refers to a plane positioning device, which can be implemented by a precision threaded screw in combination with an elastic reset structure, and is used to compensate for the alignment deviation between the light path and the flow channel.

[0060] Specifically, the sample solution is injected into the detection area of ​​the microfluidic chip 5 through the connecting pipe, and the fluid drive component 6 maintains a constant flow rate to allow the cells to pass through the light plate irradiation area in a single file. The light beam emitted by the laser light source 8 passes through the focusing component 9 to form a light plate with a thickness of about 10 μm, and a scattered light signal is generated when the cells flow through this area. The photoelectric sensor 12 of the detection component receives the light signal and converts it into an electrical pulse, and the control unit calculates the cell concentration by counting the number of pulses. The XY adjustment mechanism 11 can fine-tune the sensor position to ensure that the signal acquisition area and the light plate irradiation area precisely coincide with each other. The control unit synchronously coordinates the fluid drive, optical path stabilization and signal processing to achieve closed-loop control.

[0061] Compared to existing technologies, this device utilizes a closed fluidic system to avoid sample exposure and employs dynamic optical path adjustment technology to improve detection accuracy even at high cell concentrations. Unlike the static detection mode of traditional offline devices, the integrated fluid control and optical detection modules enable in-situ, real-time monitoring. Compared to fixed optical path designs, the adjustable detection components effectively address signal attenuation caused by optical path deviation.

[0062] In other words, this invention enables non-destructive online testing of cell samples, eliminating the risk of contamination associated with manual manipulation. A dynamic adjustment mechanism ensures stable testing of samples of varying concentrations, and a closed fluidic design supports sample recycling and recovery. The automated control system significantly improves testing efficiency and provides a reliable solution for real-time monitoring of cell culture processes.

[0063] In some embodiments of the present invention, Figure 5 、 Figure 6 As shown, the XY adjustment mechanism 11 includes an aperture assembly 13, an X-axis adjustment member 14, a Y-axis adjustment member 15, a pressure block 16, a housing bracket 17, a bracket base 18, and a pressure cover 19. The aperture assembly 13 includes an aperture 20, a lens (not labeled), and a lens pressure cover 22. The lens has a lens bracket 23. The aperture 20 and the lens pressure cover 22 are both mounted on the lens bracket 23 and are respectively located on opposite sides of the lens.

[0064] The bracket 45 and the housing 46 are fixed to the bracket base 18, and the pressing block 16 and the pressing cover 19 are respectively arranged on opposite sides of the housing bracket 17 and are fixedly connected to the housing bracket 17;

[0065] The outer shell bracket 17 is provided with a through hole 24, and one end of the aperture assembly 13 is inserted into the through hole 24 of the outer shell bracket 17. The X-axis adjustment member 14 and the Y-axis adjustment member 15 are both installed on the outer shell bracket 17, and are used to fine-tune the position of the aperture assembly 13 along the X-axis direction and the Y-axis direction, respectively.

[0066] The aperture assembly 13 is a structure composed of the aperture 20, the lens, and the lens pressure cap 22. Specifically, it can be implemented by superimposing the annular aperture 20, made of metal or polymer material, on the optical lens. The aperture 20 is used to restrict the beam shape, and the lens is used to focus the light. The X-axis adjustment member 14 is a precision displacement mechanism for horizontal movement. Specifically, it can be implemented by using a threaded fine-tuning knob in conjunction with a guide rail. Rotating the knob propels the aperture assembly 13 laterally. The Y-axis adjustment member 15 is a precision displacement mechanism for vertical movement. Specifically, it can be implemented by using a spring-loaded micrometer head structure in conjunction with a limit slot. The micrometer head's longitudinal displacement is controlled by advancing and retracting. The pressure block 16 and pressure cap 19 are clamping components used to secure the housing bracket 17. Specifically, they can be implemented by using aluminum alloy parts and fastening bolts. The symmetrically arranged clamping structure ensures the stability of the housing bracket 17.

[0067] Specifically, the aperture assembly 13 precisely aligns the aperture 20 with the lens via the lens holder 23. During assembly, the lens is first pressed into the lens holder 23, followed by the installation of the aperture 20 and the tightening of the lens cover 22. The housing holder 17 is secured to the device body via the holder base 18. The pressing block 16 and the cover 19 clamp the housing holder 17 from both sides, forming a stable support structure. When optical path calibration is required, the X-axis adjustment member 14 and the Y-axis adjustment member 15 are operated separately to cause the aperture assembly 13, inserted into the through hole 24, to undergo two-dimensional micro-movement, precisely aligning the receiving surface of the photoelectric sensor 12 with the optical path focal point. For example, the X-axis adjustment member 14 converts rotational motion into linear displacement via a threaded pair, driving the aperture assembly 13 to move horizontally within a range of 0.1-0.5 mm. The Y-axis adjustment member 15 eliminates transmission backlash through an elastic preload mechanism, enabling precise longitudinal adjustment within a range of 0.05-0.2 mm.

[0068] Compared to existing technologies, the optical path adjustment mechanisms of conventional cell counting devices often utilize a monolithic, movable structure, which cannot achieve precise adjustment in independent directions within a plane, easily leading to misalignment between the receiving surface of the photoelectric sensor 12 and the optical path. This solution utilizes a split, independent X / Y adjustment mechanism to independently correct axial deviations without removing the housing 46. This solves the problem of decreased detection sensitivity caused by cumulative assembly errors in conventional devices, while also avoiding the risk of seal damage caused by repeated disassembly and assembly.

[0069] In other words, the present invention achieves precise matching between the receiving surface of the photosensor 12 and the focal point of the optical path, effectively improving signal capture capabilities during high-concentration cell detection and reducing counting errors caused by optical path deviation. The adjustable design of the aperture assembly 13 enables the device to accommodate microfluidic chips 5 of varying sizes, enabling rapid calibration of the online detection system while maintaining a sterile operating environment.

[0070] In some embodiments of the present invention, Figure 4 As shown, it also includes an X-axis linear module 25 and a chip adjustment mechanism 26; the chip adjustment mechanism 26 is installed on the X-axis linear module 25 and can move back and forth along the X-axis direction; the tapered positioning pin (not marked in the figure) of the chip adjustment mechanism 26 cooperates with the positioning hole (not marked in the figure) on the microfluidic chip 5 to fine-tune the position of the microfluidic chip 5 along the Z-axis direction.

[0071] Among them, the X-axis linear module 25 refers to a mechanical structure that provides linear motion in the horizontal direction, which can be specifically implemented by a ball screw, a linear guide or a linear motor, and is used to drive the chip adjustment mechanism 26 to move in the X-axis direction to achieve horizontal position adjustment of the microfluidic chip 5.

[0072] Among them, the chip adjustment mechanism 26 refers to a positioning device including a conical positioning pin, which can be specifically implemented by a floating structure with an elastic element. The conical positioning pin cooperates with the positioning hole of the microfluidic chip 5 to generate a small displacement in the Z-axis direction to compensate for the installation error.

[0073] The tapered positioning pin refers to a positioning component with a tapered end, which can be made of stainless steel or carbide. Its taper design can achieve a self-centering function to ensure the alignment accuracy of the microfluidic chip 5 and the optical path unit.

[0074] Specifically, the X-axis linear module 25 drives the chip adjustment mechanism 26 to move along the X-axis, so that the conical positioning pin contacts the positioning hole of the microfluidic chip 5. When the conical positioning pin is inserted into the positioning hole, its conical surface generates contact force with the hole wall, pushing the microfluidic chip 5 to produce a slight displacement in the Z-axis direction, thereby adjusting the vertical relative position of the chip and the optical path unit. During this process, the elastic element in the chip adjustment mechanism 26 can absorb excess force to avoid chip damage due to over-positioning. In this way, the microfluidic chip 5 is precisely positioned in both the X-axis and Z-axis directions, ensuring that the cells remain vertically aligned with the laser light plate when flowing through the detection area.

[0075] Compared with existing technologies, traditional devices rely on manual chip position adjustment, which is cumbersome and has low positioning accuracy. This solution achieves automated chip position adjustment through the synergistic effect of the X-axis linear module 25 and the tapered positioning pins, eliminating human error and resolving the optical path offset problem caused by installation deviation of the microfluidic chip 5.

[0076] In other words, the present invention automatically compensates for the positioning of the microfluidic chip 5 during the cell detection process, ensuring that the laser light plate is strictly perpendicular to the cell flow path, thereby improving the stability of the signal received by the photoelectric sensor 12. This solution is particularly suitable for continuous detection of high-concentration cell samples, avoiding cell overlap errors caused by chip position deviation and reducing the need for manual intervention by operators.

[0077] In some embodiments of the present invention, Figure 1 、 Figure 7 As shown, the fluid drive assembly 6 includes a first peristaltic pump 29 and a second peristaltic pump 30. The first peristaltic pump 29 is used to pump the liquid in the test container 38 into the microfluidic chip 5; the second peristaltic pump 30 is used to pump the liquid in the dilution container 39 into the microfluidic chip 5.

[0078] The first peristaltic pump 29 is a drive device that generates directional flow by periodically squeezing an elastic tube. This can be achieved using a mechanical structure with a rotatable roller, and the liquid flow rate can be controlled by adjusting the roller speed. The second peristaltic pump 30 is a pump body with the same structure as the first peristaltic pump 29 and is independently controlled. Specifically, it can be installed in parallel to achieve dual-channel independent liquid supply. The first and second peristaltic pumps 29 and 30 drive the fluid by contacting the outer wall of the tube, avoiding direct contact between the liquid and the pump body, which is conducive to maintaining a sterile environment.

[0079] Specifically, when cell concentration testing is required, the roller of the first peristaltic pump 29 begins to rotate, squeezing the first conduit 35 to continuously deliver the cell suspension in the test container 38 to the detection channel of the microfluidic chip 5. The second peristaltic pump 30 can operate synchronously or in a time-sharing manner. For example, when testing a high-concentration sample, the second peristaltic pump 30 can pump diluent from the dilution container 39 into the detection channel for online dilution. The pinch valve 10 switches the liquid delivery path by controlling the on / off state of the third conduit 37. When sample recovery is required, the third conduit 37 is closed, allowing the liquid to flow back to the waste liquid storage device 40 through the fifth conduit 42.

[0080] Compared to existing technologies, existing offline equipment relies on manual operation for sample transfer and dilution, which is cumbersome and prone to contamination. This solution utilizes dual peristaltic pumps working together to not only automatically transfer samples but also perform dilution online, eliminating time-sensitive errors caused by manual intervention. The peristaltic pump's contactless drive keeps the pipeline closed, effectively preventing external contaminants from entering the detection system.

[0081] In other words, this invention enables automated and continuous delivery of cell suspensions, enabling online dilution of high-concentration samples while maintaining a sterile environment. The test liquid is pumped directly into the testing area via independent pipelines, minimizing the risk of sample exposure. Waste liquid can also be recycled, eliminating the waste of precious samples. The dual-pump structure synchronizes the dilution process with the testing process, allowing sample concentration to be adjusted without interrupting the testing process.

[0082] It should be noted that the online cell counting apparatus of the present invention, by providing the waste liquid storage device 40, can effectively collect waste liquid during the detection process, preventing waste liquid from contaminating the environment. The waste liquid storage device 40 is connected to the detection apparatus and the first peristaltic pump 29 via the connecting pipeline, allowing waste liquid to flow smoothly from the detection apparatus into the waste liquid storage device 40. This ensures a cleaner and safer detection process and prevents the indiscriminate discharge of waste liquid.

[0083] The installation position of the waste liquid storage device 40 can be flexibly designed, for example, it can be fixed on one side of the housing, that is, a waste liquid box 47 is set on the housing, such as Figure 1 As shown, the waste liquid storage device 40 is placed in the waste liquid box 47, wherein the waste liquid storage device 40 can be configured as a waste liquid bag. The specific location of the waste liquid storage device 40 can be adjusted according to actual needs. The capacity of the waste liquid storage device 40 can be designed based on the amount of waste liquid generated during the testing process, ensuring that the waste liquid storage device 40 is not frequently replaced during the testing process, thereby improving testing efficiency. The waste liquid storage device 40 can adopt a sealed structure to prevent volatilization and leakage of waste liquid, further ensuring a clean and safe testing environment.

[0084] By providing the waste liquid storage device 40, the online cell counting device of the present invention can effectively collect and process waste liquid during the detection process, avoiding the pollution of the waste liquid to the environment and ensuring the cleanliness and safety of the detection process. At the same time, the design of the waste liquid storage device 40 is flexible and can be adjusted according to actual needs, thereby improving the applicability and practicality of the device. Compared with the existing technology, the technical solution of the present invention has significant advantages in waste liquid treatment and can better meet the actual detection needs.

[0085] In some embodiments of the present invention, Figure 7 As shown, the microfluidic chip 5 has a cell detection channel 31 and a first inlet end 32, a second inlet end 33 and an outlet end 34 connected to the cell detection channel 31; the connecting pipeline includes a first pipeline 35, a second pipeline 36 and a third pipeline 37, one end of the first pipeline 35 is connected to the container to be tested 38, the other end of the first pipeline 35 is connected to one end of the third pipeline 37 through the connecting member 50, the other end of the third pipeline 37 is connected to the first inlet end of the microfluidic chip 5, one end of the second pipeline 36 is connected to the dilution container 39, and the other end of the second pipeline 36 is connected to the second inlet end of the microfluidic chip 5; the first peristaltic pump 29 is arranged on the first pipeline 35, the second peristaltic pump 30 is arranged on the second pipeline 36, and the pinch valve 10 is arranged on the third pipeline 37.

[0086] The cell detection channel 31 is a channel for cell flow and detection, which can be implemented by a microchannel structure with a rectangular or circular cross-section, and a width or diameter ranging from 50 to 200 microns, for example, which can constrain the single-file flow of cells.

[0087] The first inlet end 32 and the second inlet end 33 are interfaces for introducing the sample to be tested and the diluent, respectively, and can be implemented by using a tapered tapered structure, which is beneficial to reducing turbulence during fluid mixing.

[0088] The connector 50 is a component used for connecting pipelines, and can be implemented by a tee connector or a quick-connect connector, so as to facilitate switching and sealing between multiple pipelines.

[0089] Specifically, the sample to be tested is driven by the first peristaltic pump 29 through the first pipeline 35, and the diluent is driven by the second peristaltic pump 30 through the second pipeline 36. The two flow together at the connector 50 and enter the first inlet port 32 of the microfluidic chip 5 through the third pipeline 37. The pinch valve 10 regulates the flow rate of the mixed solution by controlling the on-off state of the third pipeline 37. Simultaneously, the second inlet port 33 directly receives the diluent to achieve secondary dilution. By adjusting the speed ratio of the two peristaltic pumps, the mixing ratio of the sample and diluent can be precisely controlled, allowing the high-concentration cell solution to complete a gradient dilution before flowing through the detection channel, thus avoiding counting errors caused by cell overlap.

[0090] Compared to existing offline testing methods, which require multiple manual dilutions and chip replacements, this solution achieves online automated dilution through multi-pipeline coordinated control, significantly reducing operation time while maintaining a sterile environment. Furthermore, while the existing technology relies on manual pipetting precision to determine the mixing ratio of diluent to sample, this solution achieves precise mixing with adjustable ratios through independently controlled peristaltic pumps.

[0091] In other words, this invention solves the problem of repeated manual dilution required for high-concentration cell testing, avoiding the risk of sample exposure and contamination. It also improves detection accuracy through a multi-stage dilution channel design. The dynamic adjustment of the mixing ratio also allows for adaptability to different cell concentrations, enhancing the device's versatility.

[0092] In some embodiments of the present invention, Figure 7 As shown, it also includes a waste liquid storage device 40, and the connecting pipeline also includes a fourth pipeline 41 and a fifth pipeline 42. One end of the fourth pipeline 41 and the fifth pipeline 42 are both connected to the waste liquid storage device 40, and the other end of the fourth pipeline 41 is connected to the outlet end 34 of the microfluidic chip 5. The other end of the fifth pipeline 42 is connected to the first pipeline 35 and the third pipeline 37 through the connecting piece 50; the pinch valve 10 is a two-way pinch valve 10, and the opening and closing of the third pipeline 37 and the fifth pipeline 42 are both controlled by the pinch valve 10.

[0093] Among them, the waste liquid storage device 40 refers to a container for collecting waste liquid generated during the detection process, which can be implemented by a sealed liquid storage tank to prevent waste liquid leakage from causing environmental pollution. The fourth pipeline 41 refers to a transmission channel connecting the outlet end 34 of the microfluidic chip 5 and the waste liquid storage device 40, which can be implemented by a flexible silicone tube to ensure smooth discharge of waste liquid. The fifth pipeline 42 refers to an auxiliary channel connecting the waste liquid storage device 40 with the first pipeline 35 and the third pipeline 37, which can be implemented by a three-way connector to achieve multi-directional fluid communication. The two-way clamping valve 10 refers to a valve that can control the on and off of two pipelines at the same time, which can be implemented by an electromagnetically driven clamping valve 10, and the synchronous opening and closing action is achieved through the control circuit.

[0094] Specifically, during the cell testing process, when sample recovery is required, the pinch valve 10 simultaneously closes the third line 37 and opens the fifth line 42, allowing the original sample in the first line 35 to flow directly back to the waste liquid storage device 40 through the fifth line 42 for temporary storage. When waste liquid discharge is required, the pinch valve 10 closes the fifth line 42 and opens the third line 37, allowing the waste liquid after testing to be discharged into the waste liquid storage device 40 through the fourth line 41. This dual-line design creates independent channels for sample recovery and waste liquid discharge, allowing rapid switching between the two operating modes using a single valve.

[0095] Compared to existing technologies, traditional cell counting devices typically utilize an open waste tank with only a single discharge channel, which prevents sample recovery and poses a risk of cross-contamination. By adding the fifth pipeline 42 and the two-way pinch valve 10, this solution achieves the dual functions of waste liquid discharge and sample recovery while maintaining a compact piping system. In particular, the synchronized control feature of the two-way pinch valve 10 avoids the complexity of traditional solutions requiring coordinated operation of multiple valves.

[0096] In other words, the present invention effectively solves the technical problem of rare cell sample recovery. Unused raw samples can be safely stored during the testing process, avoiding sample waste caused by traditional offline testing. The dual-pipeline shunt design also significantly reduces the risk of cross-contamination between different fluids, making it particularly suitable for continuous testing operations in sterile environments. The integrated use of the two-way pinch valve 10 further simplifies the device's operating procedures, providing the online cell counting device 100 with a higher level of automation.

[0097] In some embodiments of the present invention, a device body (not marked in the figure) is also included, such as Figure 1-Figure 4As shown, the device body includes a base plate 44, a bracket 45 and a shell 46. The bracket 45 is installed on the base plate 44, the detection component is installed on the bracket 45, the bottom surface of the shell 46 is fixed to the base plate 44, and the fluid drive component 6 and the switch component are both installed on the top surface of the shell 46.

[0098] The bottom plate 44 refers to a supporting structure for carrying the internal components of the device body, and can be specifically implemented by a metal plate or a composite material plate, and is used to provide rigid support for the bracket 45 and the housing 46 .

[0099] The bracket 45 refers to a support frame vertically mounted on the base plate 44 , and can be made of aluminum alloy profiles or injection molding structures, and is used to fix the detection component and maintain its spatial position stability.

[0100] The housing 46 refers to a protective shell covering the internal structure of the device body, and can be specifically implemented by split injection molding of the housing 46 or sheet metal splicing of the housing 46 to isolate the internal components from interference from the external environment.

[0101] The switch element refers to an actuator that controls the on / off of a pipeline, and can be implemented by a solenoid valve, the pinch valve 10 or a rotary valve, and is used to adjust the liquid flow path.

[0102] Specifically, the base plate 44 is rigidly connected to the bracket 45 by bolts or welding, the detection component is fixed to the preset mounting position of the bracket 45 by screws or clips, and the housing 46 is sealed to the base plate 44 by edge slots. The fluid drive component 6 and the switch are installed in the reserved mounting area on the top surface of the housing 46 by guide rails or locating pins, and their control cables are connected to the internal circuit through the wire holes in the side walls of the housing 46. As a result, the main body of the device forms a layered layout structure, with the base plate 44 assuming mechanical support, the bracket 45 achieving precise positioning of the detection component, and the housing 46 providing physical protection and integrating the external operation interface.

[0103] Compared to existing cell counting devices, which typically use a split structure, the fluid drive component and the detection component are installed separately, resulting in bulky equipment and difficult maintenance. This solution integrates the device body design, centrally arranging the fluid drive assembly 6 and switch components on the top surface of the housing 46, making it easier for operators to quickly inspect and replace components. Furthermore, the rigid connection between the bracket 45 and the base plate 44 ensures the positioning accuracy of the detection assembly.

[0104] In other words, the present invention achieves a compact and modular structure for the device's main structure, resolving the bulk and inconvenient maintenance issues of existing devices. The rigid connection between the base plate 44 and the bracket 45 ensures the stable positioning of the detection assembly. The sealed design of the housing 46 reduces the risk of external contamination. The top-surface layout of the fluid drive assembly 6 and the switch element optimizes operational convenience, thereby supporting the stable operation of the online cell counting device 100 in a continuous production environment.

[0105] In some embodiments of the present invention, Figure 3 、 Figure 4 As shown, the laser light source 8 is mounted on the bracket 45, the X-axis linear module 25 is mounted on the base plate 44 below the bracket 45, the bracket 45 is provided with the chip slot, the chip slot is located at the end of the focusing assembly 9 away from the laser light source 8, the microfluidic chip 5 is mounted in the chip slot, the liquid path unit is connected to the dilution container 39, the test container 38 and the waste liquid storage device 40, and the laser light source 8, the focusing assembly 9, the microfluidic chip 5, the aperture assembly 13 and the photoelectric sensor 12 are located on the same straight line.

[0106] The laser light source 8 is a device for generating an excitation light beam required for detection, which can be implemented by a semiconductor laser. The wavelength range of the laser light source 8 can be set to 405-650 nm, for example, to excite cells flowing through the microfluidic chip 5 to generate light signals.

[0107] Among them, the X-axis linear module 25 refers to a mechanical structure that drives the microfluidic chip 5 to perform horizontal displacement, which can be specifically achieved by using a stepper motor and a ball screw transmission method to adjust the horizontal position of the chip during the detection process to adapt to different detection areas.

[0108] Among them, the chip slot refers to a positioning structure for fixing the microfluidic chip 5, which can be specifically implemented by an aluminum alloy groove with an elastic snap. The inner wall of the groove can be provided with the conical positioning pin to cooperate with the positioning hole of the chip to achieve precise positioning of the chip in the vertical direction.

[0109] Among them, the liquid circuit unit connects the dilution container 39, the test container 38 and the waste liquid storage device 40, which refers to a pipeline connection method for constructing a closed fluid circulation system. Specifically, it can be achieved by combining a medical-grade silicone tube and a Luer lock connector to realize automatic dilution, transportation and waste liquid recovery of the sample.

[0110] Specifically, the excitation light beam emitted by the laser light source 8 passes through the focusing component 9 to form a light plate with a thickness of 10-50 μm, which vertically penetrates the detection channel of the microfluidic chip 5. When the liquid containing cells flows through the detection area, the cells passing through the light plate area will trigger scattered light and fluorescence signals, which are captured by the photoelectric sensor 12 and converted into electrical signals. The X-axis linear module 25 drives the chip card slot to move horizontally, so that different detection areas are aligned with the optical path system in turn. The liquid path unit is controlled by a peristaltic pump to realize automatic dilution and transportation of samples, and the waste liquid storage device 40 is connected by an airtight pipeline to ensure the sterility of the operation process. The design of arranging the optical elements along the same straight line reduces the optical path transmission loss to less than 5%.

[0111] Compared to existing technologies, traditional cell counters require manual chip placement and are unable to achieve continuous detection. This solution, however, achieves online continuous detection of samples through an integrated fluidic system and automatic positioning mechanism. Existing devices often use a discrete optical system, resulting in bulky designs. This solution integrates the optical components onto a single bracket 45 through a linear optical path layout, reducing the device size by approximately 40%. Existing technologies require manual dilution when processing high-concentration samples. This solution, through the automatic dilution function of the fluidic unit, dynamically adjusts the sample concentration to an appropriate detection range of 1E5-1E7 cells / ml.

[0112] In other words, the present invention automates the entire cell counting process, eliminating the risk of contamination associated with manual operation. The closed fluidic system maintains sample sterility, making it particularly suitable for bioreactor applications requiring continuous monitoring. The collinear layout of the optical components effectively improves signal acquisition efficiency, enabling detection sensitivity to reach the single-cell level. The modular design of the X-axis linear module 25 and the chip slot structure enables the device to accommodate detection chips of varying specifications, expanding its adaptability to various application scenarios.

[0113] In some embodiments of the present invention, Figure 3 、 Figure 4 As shown, the control unit includes a control circuit board 48 and a display screen 49. The display screen 49 is mounted on the top surface of the housing 46. The control circuit board 48 is mounted on the bracket 45. The control circuit board 48 is electrically connected to the detection component, the moving component, the fluid drive component 6, the pinch valve 10 and the display screen 49.

[0114] The control circuit board 48 refers to an integrated circuit board for receiving and processing electrical signals and outputting control instructions. Specifically, it can be implemented by a multi-layer PCB board integrating a microprocessor and a drive circuit, and is used to coordinate the timing of the actions of various execution components. The display screen 49 refers to a visual output device for human-computer interaction, and can be implemented by a touch-screen LCD screen, and is used to display cell counting data and system status parameters in real time. The top surface of the housing 46 refers to the flat area of ​​the shell covering the upper part of the device body, and can be made of metal or engineering plastics to support the display screen 49 and form a sealed structure. The bracket 45 refers to a rigid frame structure that supports the detection component, and can be formed by welding aluminum alloy profiles to fix the control circuit board 48 and maintain its spatial position stability.

[0115] Specifically, the control circuit board 48 establishes a signal transmission channel with the photoelectric sensor 12 via wires, receiving in real time the pulse signals generated by cells passing through the microfluidic chip 5. The control circuit board 48 generates the drive signal for the stepper motor of the moving assembly, which adjusts the displacement of the X-axis linear module 25 to achieve chip positioning. The control circuit board 48 outputs the peristaltic pump speed command for the fluid drive assembly 6, controlling the liquid flow rate by varying the pulse frequency. The opening and closing states of the pinch valve 10 are switched by high and low level signals sent by the control circuit board 48, achieving precise control of the pipeline's opening and closing. The display screen 49 is connected to the control circuit board 48 via a cable and dynamically updates the processed cell concentration data in graphical form. The tilt angle of the top surface of the housing 46 has been ergonomically calculated, allowing the display screen 49 to face the operator at a 15-degree elevation angle. Wiring channels are provided within the bracket 45 to systematically route the cables connecting the control circuit board 48 to the various actuators.

[0116] In some embodiments, the control circuit board 48 can be divided into independent functional areas, such as a signal acquisition area equipped with an A / D converter chip, a motion control area equipped with a stepper motor driver chip, and a fluid control area integrated with a MOS transistor switching circuit. The display screen 49 can be mounted using a magnetic quick-release structure for quick replacement during maintenance. The bracket 45 and the control circuit board 48 are preferably secured with insulating nylon posts and screws to prevent the risk of short circuits.

[0117] Compared with existing technologies, the control modules of traditional cell counters are usually arranged in a dispersed manner, resulting in bulky devices and difficulty in heat dissipation. By integrating the control circuit board 48 within the bracket 45, this solution significantly shortens the signal transmission distance between the sensor and the processor, effectively reducing electromagnetic interference. The integrated design of the display screen 49 and the top surface of the housing 46 replaces the connection cables of traditional external displays, making the operation interface layout more compact and reasonable. The direct connection between the control circuit board 48 and the actuator abandons the traditional relay control mode, improving the system's response speed and reliability.

[0118] In other words, the present invention achieves centralized processing of control commands from multiple subsystems, ensuring precise synchronization of actions such as fluid actuation, optical detection, and mechanical positioning. Real-time data feedback from the display screen 49 enables the operator to intuitively monitor changes in cell concentration, avoiding detection errors caused by information lag. The integrated mounting of the control circuit board 48 and the bracket 45 optimizes internal space utilization, leaving ample space for the heat dissipation components, thereby meeting the stability requirements for long-term continuous operation.

[0119] In some embodiments of the present invention, Figure 3 、 Figure 4 As shown, a heat dissipation component is also included, and the heat dissipation component (not marked in the figure) includes a first heat dissipation member 51, a second heat dissipation member 52 and a third heat dissipation member 53. The first heat dissipation member 51 is installed on the laser light source 8, and the second heat dissipation member 52 and the third heat dissipation member 53 are both installed on the base plate 44, and the second heat dissipation member 52 is located in the middle of the base plate 44, and the third heat dissipation member 53 is located at the edge of the base plate 44.

[0120] The first heat sink 51 is a heat dissipation component in direct contact with the laser light source 8, and can be implemented as a metal heat sink or a micro fan. It is used to quickly dissipate the heat generated by the laser light source 8. The second heat sink 52 is a heat dissipation structure located in the middle of the base plate 44, and can be implemented as heat dissipation fins or heat pipes. It is used to balance the concentrated heat generated during device operation. The third heat sink 53 is an auxiliary heat dissipation component located at the edge of the base plate 44, and can be implemented as a heat dissipation grille or heat dissipation holes. It is used to enhance air circulation efficiency in the edge area of ​​the device.

[0121] Specifically, the laser light source 8 generates a large amount of heat when working for a long time. The first heat sink 51 conducts the heat to the external environment through physical contact. The middle part of the base plate 44 is prone to forming a heat accumulation area because it carries components such as the fluid drive component 6 and the control unit. The second heat sink 52 accelerates the heat diffusion in this area by increasing the heat dissipation area. The edge of the base plate 44 is prone to forming a heat dissipation dead corner because it is close to the enclosed area of ​​the shell 46. The third heat sink 53 improves the heat dissipation efficiency of the edge area by optimizing the airflow path. The three groups of heat sinks are differentiated according to the distribution characteristics of different heat sources to form a coordinated heat dissipation mechanism.

[0122] Compared to existing technologies, traditional cell counting devices typically only have a single heat dissipation structure in key components, which is unable to cope with the combined heat load generated by the simultaneous operation of the high-power laser light source 8, the fluid drive assembly 6, and the control unit. This solution, by providing a zoned heat dissipation structure, can provide targeted heat dissipation for the laser light source 8, the core area of ​​the device, and the edge areas, effectively preventing problems such as optical path deviation, fluid parameter fluctuations, and electronic component performance degradation caused by localized overheating.

[0123] In other words, this invention solves the problem of uneven heat dissipation in existing cell counting devices, which leads to reduced detection accuracy and insufficient device stability. The zoned heat dissipation design maintains the stability of the laser wavelength in the optical path unit, preventing fluctuations in the fluid-driven flow rate caused by temperature changes. It also extends the life of the control unit's electronic components, ensuring data reliability during the cell counting process.

[0124] Furthermore, if Figure 2 As shown, a plurality of vents 7 may be provided on the housing to further accelerate the air circulation inside the housing, thereby improving the working efficiency of the heat dissipation component and improving the heat dissipation capacity.

[0125] In some embodiments of the present invention, Figure 2 As shown, it also includes a power socket 1, which is installed on the bracket. A first hole is provided on the shell, and the first hole corresponds to the power socket 1. The power socket 1 is electrically connected to the detection component, the control circuit board and the fluid drive component 6.

[0126] The provision of the power socket 1 ensures stable power supply to the device. By installing the power socket 1 on the bracket and providing a corresponding first hole on the housing, power connection is made more convenient, and the exposure of wires can be effectively avoided, thereby improving the overall aesthetics and safety of the device. Furthermore, a power switch and the indicator light can be provided on the bracket. The power switch and the indicator light are electrically connected to the power socket 1 and are exposed to the external environment through the housing. The power switch can further control the power supply of the device, thereby improving the circuit safety of the device; the provision of the indicator light can indicate the circuit status, thereby improving the convenience of using the device.

[0127] The power socket 1 can be a standard power socket or other interfaces that can connect to the power supply, such as a USB interface 21. The power socket 1 is installed at an appropriate position on the bracket and is exposed to the external environment through the first hole on the shell. The first hole can be set with a sealed design to prevent the external environment from affecting the circuit. The power socket 1 is connected to the detection component, the control circuit board and the drive component 8 through wires to ensure that these components can work stably. As a preferred embodiment, the power socket 1 can adopt a quick-plug design to facilitate users to quickly connect and disconnect the power supply.

[0128] The above design makes the online cell counting device of the present invention more convenient in terms of power connection, improving the user experience and safety of the device. Compared with the existing technology, the solution of the present invention effectively solves the problem of inconvenient power connection, ensures stable power supply of the device, and improves the overall reliability and aesthetics of use.

[0129] Furthermore, if Figure 1 As shown, the housing is further provided with a power switch 2, which is electrically connected to the power socket 1 to achieve power control of the device and further improve the safety and stability of the power supply to the device.

[0130] In some embodiments of the present invention, Figure 2 As shown, it also includes a network socket 3, which is installed on the bracket. A second hole (not marked in the figure) is provided on the shell, and the second hole is set corresponding to the network socket 3. The network socket 3 is electrically connected to the detection component, the control circuit board and the fluid drive component 6.

[0131] The network port 3 enables data transmission and remote control of the online cell counter via a network interface. Specifically, the network port 3 connects to the network, enabling the detection component, the control circuit board, and the driver component to communicate with an external computer or server, thereby enabling real-time data transmission and remote monitoring. This allows users to remotely access and control the online cell counter via the network, obtain real-time detection data, and perform corresponding operations and adjustments.

[0132] As a preferred embodiment, the network socket 3 can adopt a standard Ethernet interface, such as an RJ45 interface, to ensure compatibility with existing network devices. Further, the network socket 3 can be protected by a shielding cover to prevent external interference and physical damage.

[0133] By adding the network port 3 to the device, the present invention effectively solves the problem of existing offline cell counters requiring manual operation and inconvenient data transmission. Compared with existing technologies, the present invention has the advantages of enabling online real-time data transmission and remote control, improving detection efficiency and data reliability, and reducing the risk of human interference and contamination.

[0134] Furthermore, if Figure 2 As shown, the online cell counting device also includes a connection port 4, which is electrically connected to an external device via a connection line, thereby enabling the online cell counting device to be controlled by the external device, or enabling the online cell counting device to be analyzed and counted by the external device. Furthermore, the connection port 4 can also be connected to other external devices to achieve more functions, which are not detailed here.

[0135] In addition, if Figure 1 As shown, the online cell counting device further includes a USB interface 21, which is used to connect to an external device. Specifically, by providing the USB interface 21, an external power source can be connected instead of the power socket 1. By providing a socket type different from the power socket 1, the online cell counting device can be used in different environments. The USB interface 21 can also replace the connection port 4 for connecting to an external device, allowing the online cell counting device to be connected to multiple external devices at the same time. The versatility of the online cell counting device is improved by providing an interface of a different type from the connection port 4.

[0136] The present invention also provides a method for controlling online cell counting, which includes the online cell counting device 100 as described above. Figure 8 As shown, the control method includes:

[0137] Step S1: providing the online cell counting device 100;

[0138] Step S2: connecting the test container 38, the dilution container 39 and the waste liquid storage device 40 to the online cell counting device 100;

[0139] Step S3: Inserting the microfluidic chip 5:

[0140] Step S4: turning on the first peristaltic pump 29 to allow the cell fluid to fill the connecting pipe and rinse the connecting pipe;

[0141] Step S5: selecting a counting mode according to the cell concentration and starting online cell counting detection;

[0142] Step S6: transporting the detected cells to the waste liquid storage device 40 for collection and processing.

[0143] Specifically, the power supply is connected to the power port, and a network cable is connected to the network socket 3, so that the device can communicate with external devices through the network; the microfluidic chip 5 is inserted into the chip mounting part 130, and the microfluidic chip 5 is connected to a port of the switch component through the connecting pipe, and then connected to the container to be tested 38, wherein the first peristaltic pump 29 is connected between the switch component and the container to be tested 38 through the connecting pipe; the microfluidic chip 5 is connected to the dilution container 39, wherein the second peristaltic pump 30 is connected between the microfluidic chip 5 and the dilution container 39 through the connecting pipe; the microfluidic chip 5 is directly connected to the waste liquid storage device 40 through the connecting pipe; the waste liquid storage device 40 is directly connected to the container to be tested 38 through the switch component.

[0144] That is, the online cell counting device 100 is first connected to the power supply and the network, the microfluidic chip 5 is installed, the container to be tested 38, the dilution container 39 and the waste liquid storage device 40 are connected to the online cell counting device 100, the power switch 2 is pressed to turn on the online cell counting device 100, the control circuit board 48 is operated to control the switch element 6 to connect the waste liquid storage device 40 and the container to be tested 38, and the first peristaltic pump 29 is controlled to transport the cell fluid to be tested from the container to be tested 38 to the waste liquid storage device 40 to rinse the connecting pipeline, thereby preventing the residual substances in the connecting pipeline from affecting the experimental results and improving the accuracy of the detection; further, the control circuit board 48 is operated to control the switch element to close the waste liquid storage device 40. The liquid storage device 40 is connected to the container to be tested 38, and the container to be tested 38 and the microfluidic chip 5 are connected to control the first peristaltic pump 29 to transport the cell fluid to be tested to the microfluidic chip 5, and then operate the control circuit board 48 to turn on the laser light source 8, adjust the focusing component 9, the XY adjustment mechanism 11 and the microfluidic chip 5 to the appropriate position, so that the laser passes through the focusing channel of the microfluidic chip 5, and the scattered light beam is collected and blocked by the XY adjustment mechanism 11. Further, the scattered light is converted into an electrical signal and processed by an algorithm to perform cell counting and cell size analysis; after the cell counting test is completed, the first peristaltic pump 29 is controlled to transport the tested cell fluid to the waste liquid storage device 40 to collect and process the waste liquid.

[0145] In some embodiments of the present invention, Figure 9 As shown, a counting mode is selected according to the cell concentration, and online cell counting detection is started, including:

[0146] Step S51: detecting or inputting the cell concentration, and selecting a counting mode according to the cell concentration;

[0147] Step S52: If the low concentration counting mode is selected, the fluid driving component 6 is controlled to turn on the first peristaltic pump 29 to transport the cells to be tested to the microfluidic chip 5;

[0148] If the high concentration counting mode is selected, the first peristaltic pump 29 and the second peristaltic pump 30 are turned on to deliver the cells to be tested to the microfluidic chip 5 while delivering the diluent into the microfluidic chip 5 to adjust the concentration of the cells to be tested;

[0149] Step S53: turning on the laser light source 8 to allow the laser to pass through the focusing assembly 9 and enter the focusing detection channel of the microfluidic chip 5;

[0150] Step S54: adjusting the height of the microfluidic chip 5 so that the light passing through the detection channel is incident on the sensing surface of the photoelectric sensor 12, thereby achieving cell counting.

[0151] Specifically, the normal concentration detection range of the microfluidic chip 5 is <5E7 / ml. When the cell fluid concentration in the test container 38 satisfies <5E7 / ml, the second peristaltic pump 30 is not turned on, and the online cell counting device 100 performs cell detection normally. When the cell fluid concentration in the test container 38 is >5E7 / ml, the second peristaltic pump 30 is controlled to deliver the diluent to the microfluidic chip 5 to dilute the test cell fluid so that the concentration of the test cell fluid satisfies <5E7 / ml, thereby ensuring accurate counting of the microfluidic chip 5.

[0152] It should be understood that the above-described specific embodiments of the present invention are merely illustrative or illustrative of the principles of the present invention and do not constitute limitations of the present invention. Therefore, any modifications, equivalent substitutions, improvements, etc. made without departing from the spirit and scope of the present invention should be included within the scope of protection of the present invention. In addition, the appended claims are intended to cover all variations and modifications that fall within the scope and metes and bounds of the appended claims, or equivalents thereof.

Claims

1. An online cell counting device, characterized in that: include: A fluid circuit unit, comprising a microfluidic chip, a fluid drive component, and a connecting pipeline, wherein the connecting pipeline is connected to the microfluidic chip, and the fluid drive component is disposed on the connecting pipeline to drive the fluid to flow in the microfluidic chip; An optical path unit, comprising a laser light source and a focusing assembly, wherein the outgoing light of the laser light source is focused into a light plate by the focusing assembly and then incident on the microfluidic chip; A detection component is disposed on a side of the microfluidic chip facing away from the focusing component; a control unit, configured to control the working states of the fluid drive assembly, the pinch valve on the connecting pipeline, and the detection assembly, and to obtain a cell counting result based on an electrical signal provided by the detection assembly; The detection component includes an XY adjustment mechanism and a photoelectric sensor installed on the XY adjustment mechanism. The XY adjustment mechanism is used to fine-tune the position of the photoelectric sensor along the X-axis direction and the Y-axis direction respectively.

2. The online cell counting device according to claim 1, characterized in that The XY adjustment mechanism includes an aperture assembly, an X-axis adjustment member, a Y-axis adjustment member, a pressure block, a housing bracket, a bracket base, and a pressure cover; The aperture assembly includes an aperture, a lens, and a lens gland. The lens has a lens holder. The aperture and the lens gland are both mounted on the lens holder and are located on opposite sides of the lens. The bracket shell is fixed on the bracket base, and the pressing block and the pressing cover are respectively arranged on opposite sides of the shell bracket and are both fixedly connected to the shell bracket; The outer shell bracket is provided with a through hole, one end of the aperture assembly is inserted into the through hole of the outer shell bracket, and the X-axis adjustment member and the Y-axis adjustment member are both installed on the outer shell bracket, and are used to fine-tune the position of the aperture assembly along the X-axis direction and the Y-axis direction respectively.

3. The online cell counting device according to claim 1, characterized in that: It also includes an X-axis linear module and a chip adjustment mechanism; The chip adjustment mechanism is installed on the X-axis linear module and can move back and forth along the X-axis direction; the tapered positioning pins of the chip adjustment mechanism cooperate with the positioning holes on the microfluidic chip to fine-tune the position of the microfluidic chip along the Z-axis direction.

4. The online cell counting device according to claim 1, characterized in that The fluid drive assembly includes a first peristaltic pump and a second peristaltic pump. The first peristaltic pump is used to pump the liquid in the container to be tested into the microfluidic chip; the second peristaltic pump is used to pump the liquid in the dilution container into the microfluidic chip.

5. The online cell counting device according to claim 4, characterized in that: The microfluidic chip has a cell detection channel and a first inlet end, a second inlet end and an outlet end connected to the cell detection channel; the connecting pipeline includes a first pipeline, a second pipeline and a third pipeline, one end of the first pipeline is connected to the container to be tested, the other end of the first pipeline is connected to one end of the third pipeline through a connector, the other end of the third pipeline is connected to the first inlet end of the microfluidic chip, one end of the second pipeline is connected to the dilution container, and the other end of the second pipeline is connected to the second inlet end of the microfluidic chip; the first peristaltic pump is arranged on the first pipeline, the second peristaltic pump is arranged on the second pipeline, and the pinch valve is arranged on the third pipeline.

6. The online cell counting device according to claim 5, characterized in that: It also includes a waste liquid storage device, and the connecting pipeline also includes a fourth pipeline and a fifth pipeline. One end of the fourth pipeline and the fifth pipeline are both connected to the waste liquid storage device, the other end of the fourth pipeline is connected to the outlet end of the microfluidic chip, and the other end of the fifth pipeline is connected to the first pipeline and the third pipeline through the connecting piece; the pinch valve is a two-way pinch valve, and the opening and closing of the third pipeline and the fifth pipeline are both controlled by the pinch valve.

7. The online cell counting device according to claim 1, characterized in that: It also includes a device body, which includes a base plate, a bracket and a shell. The bracket is installed on the base plate, the detection component is installed on the bracket, the bottom surface of the shell is fixed to the base plate, and the fluid drive component and the switch are both installed on the top surface of the shell.

8. The online cell counting device according to claim 7, characterized in that: The laser light source is mounted on the bracket, the X-axis linear module is mounted on the base plate below the bracket, a chip slot is provided on the bracket, the chip slot is located at the end of the focusing assembly away from the laser light source, the microfluidic chip is mounted in the chip slot, the liquid path unit is connected to the dilution container, the container to be tested and the waste liquid storage device, and the laser light source, the focusing assembly, the microfluidic chip, the aperture assembly and the photoelectric sensor are located on the same straight line.

9. The online cell counting device according to claim 7, characterized in that: The control unit includes a control circuit board and a display screen. The display screen is mounted on the top surface of the housing. The control circuit board is mounted on the bracket. The control circuit board is electrically connected to the detection component, the moving component, the fluid drive component, the pinch valve and the display screen.

10. The online cell counting device according to claim 7, characterized in that: It also includes a heat dissipation component, which includes a first heat dissipation component, a second heat dissipation component and a third heat dissipation component. The first heat dissipation component is installed on the laser light source, and the second heat dissipation component and the third heat dissipation component are both installed on the base plate, and the second heat dissipation component is located in the middle of the base plate, and the third heat dissipation component is located at the edge of the base plate.

11. A method for controlling online cell counting, characterized in that: include: Step S1: providing an online cell counting device as described in any one of claims 1 to 10; Step S2: connecting the container to be tested, the dilution container, and the waste liquid storage device to the online cell counting device; Step S3: Inserting the microfluidic chip: Step S4: turning on the first peristaltic pump to allow the cell fluid to fill the connecting pipeline to rinse the connecting pipeline; Step S5: selecting a counting mode according to the cell concentration and starting online cell counting detection; Step S6: transporting the detected cells to the waste liquid storage device for collection and processing.

12. The online cell counting control method according to claim 11, characterized in that: Select a counting mode according to the cell concentration and start online cell counting detection, including: Step S51: detecting or inputting the cell concentration, and selecting a counting mode according to the cell concentration; Step S52: If the low concentration counting mode is selected, controlling the fluid drive assembly to turn on the first peristaltic pump to transport the cells to be tested to the microfluidic chip; If the high concentration counting mode is selected, the first peristaltic pump and the second peristaltic pump are turned on to deliver the cells to be tested to the microfluidic chip, and at the same time, a diluent is delivered into the microfluidic chip to adjust the concentration of the cells to be tested; Step S53: turning on the laser light source to allow the laser to pass through the focusing assembly and enter the focusing detection channel of the microfluidic chip; Step S54: adjusting the height of the microfluidic chip so that the light passing through the focused detection channel is incident on the sensing surface of the photoelectric sensor, thereby achieving cell counting.

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