An online cell counting device and control method

Abstract

The application relates to the technical field of cell counting, and provides an online cell counting device and a control method, the online cell counting device comprising a device main body, a detection assembly, a fluid driving assembly and a connecting pipeline, the detection assembly and the fluid driving assembly being installed on the device main body, the detection assembly being communicated with the fluid driving assembly through the connecting pipeline, and the fluid driving assembly being communicated with a to-be-detected container through the connecting pipeline and enabling to-be-detected cells in the to-be-detected container to enter the detection assembly. The cells in the to-be-detected container are directly transported into the detection assembly for detection through the fluid driving assembly, online real-time monitoring is realized, the manual sampling process is avoided, the sterile environment is ensured, and meanwhile, through cooperation of the connecting pipeline and the fluid driving assembly, accurate detection of high-concentration cells and cell recycling can be realized.

Description

Technical Field

[0001] This invention relates to the field of cell counting technology, and more specifically, to an online cell counting device and control method. Background Technology

[0002] Most existing cell counters are offline, such as the Thermo Countess 3. This type of offline cell counter requires manual removal of cells, placement into a detection chip, and then the chip being placed into the instrument for image capture and identification. This method has the following problems: First, the cell removal process is time-sensitive and cannot reflect the cell concentration and state during the encapsulation process in real time. Second, after removal, the cells are exposed to the external environment, posing a risk of contamination and compromising the sterile environment.

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

[0004] Another noteworthy issue is that existing offline cell counting methods cannot effectively recover some rarer cells. This not only wastes valuable samples but also limits the research and application of these cells.

[0005] Existing cell counting technologies also have other limitations. For example, they lack automation and real-time monitoring capabilities, failing to meet the online monitoring needs of continuous production processes. Furthermore, existing technologies lack flexibility and adaptability when handling different types and sizes of cells, making it difficult to meet diverse research and production requirements.

[0006] Furthermore, existing cell counting devices are often bulky and difficult to integrate into other systems, limiting their application in complex experimental or production environments. At the same time, operating these devices often requires specialized training, increasing the cost and barrier to entry for their use.

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

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

[0009] To achieve the above objectives, the present invention adopts the following technical solution: An online cell counting device according to an embodiment of the present invention includes: a fluid path unit, including a microfluidic chip, a fluid driving component, and a connecting pipe, wherein the connecting pipe is connected to the microfluidic chip, and the fluid driving component is disposed on the connecting pipe for driving fluid to flow in the microfluidic chip; an optical path unit, including a laser source and a focusing component, wherein the emitted light from the laser source is focused into a light plate by the focusing component and then incident on the microfluidic chip; a detection component, disposed on the side of the microfluidic chip opposite to the focusing component; and a control unit, used to control the working state of the fluid driving component, the clamp valve on the connecting pipe, and the detection component, and to obtain cell counting results based on the electrical signal provided by the detection component; the detection component includes an XY adjustment mechanism and a photoelectric sensor mounted on the XY adjustment mechanism, wherein the XY adjustment mechanism is used to finely adjust the position of the photoelectric sensor along the X-axis and Y-axis directions, respectively.

[0010] The online cell counting device according to embodiments of the present invention constructs a closed-loop control system by integrating a liquid path unit, an optical path unit, a detection component, and a control unit. It uses a microfluidic chip to realize the automatic delivery and detection of cell samples, and combines an adjustable photoelectric sensor positioning mechanism to ensure detection accuracy. It has the advantages of realizing real-time online monitoring of the cell culture process, avoiding 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 improving the system integration and automation level.

[0011] In addition, the online cell counting device according to the above embodiments of the present invention may also have the following additional technical features: 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 having a lens bracket, the aperture and the lens pressure cover being mounted on the lens bracket and located on opposite sides of the lens; the housing bracket is fixed on the bracket base, the pressure block and the pressure cover being disposed on opposite sides of the housing bracket and fixedly connected to the housing bracket; the housing bracket has a through hole, one end of the aperture assembly being inserted into the through hole of the housing bracket, the X-axis adjustment member and the Y-axis adjustment member being mounted on the housing bracket and used to finely adjust the position of the aperture assembly along the X-axis and Y-axis directions, respectively.

[0012] 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 mounted on the X-axis linear module and can move back and forth along the X-axis direction; the tapered positioning pin of the chip adjustment mechanism cooperates with the positioning hole on the microfluidic chip to finely adjust the position of the microfluidic chip along the Z-axis direction.

[0013] 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 liquid in the test container into the microfluidic chip; and the second peristaltic pump is used to pump liquid in the dilution container into the microfluidic chip.

[0014] 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 communicating with 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 test container, the other end of the first pipeline is connected to one end of the third pipeline via 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; a first peristaltic pump is disposed on the first pipeline, a second peristaltic pump is disposed on the second pipeline, and the clamp valve is disposed on the third pipeline.

[0015] In some embodiments of the present invention, a waste liquid storage device is also included. The connecting pipeline further includes a fourth pipeline and a fifth pipeline. One end of each of the fourth and fifth pipelines 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. The other end of the fifth pipeline is connected to the first pipeline and the third pipeline through the connector. The pinch valve is a two-way pinch valve, and the opening and closing of the third and fifth pipelines are controlled by the pinch valve.

[0016] In some embodiments of the present invention, the device body is further included, the device body including a base plate, a bracket and a housing, the bracket being mounted on the base plate, the detection component being mounted on the bracket, the bottom surface of the housing being fixed to the base plate, and the fluid drive component and the switch being mounted on the top surface of the housing.

[0017] In some embodiments of the present invention, the laser source is mounted on the bracket, the X-axis linear module is mounted on the base plate below the bracket, the bracket is provided with a chip slot, the chip slot is located at the end of the focusing component away from the laser source, the microfluidic chip is mounted in the chip slot, the liquid circuit unit connects the dilution container, the test container and the waste liquid storage device, and the laser source, the focusing component, the microfluidic chip, the aperture component and the photoelectric sensor are located on the same straight line.

[0018] In some embodiments of the present invention, the control unit includes a control circuit board and a display screen, the display screen being mounted on the top surface of the housing, the control circuit board being mounted on the bracket, and the control circuit board being electrically connected to the detection component, the movement component, the fluid drive component, the clamp valve, and the display screen.

[0019] In some embodiments of the present invention, a heat dissipation assembly is further included, which includes a first heat dissipation component, a second heat dissipation component, and a third heat dissipation component. The first heat dissipation component is mounted on the laser light source, and the second heat dissipation component and the third heat dissipation component are both mounted on a base plate. 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.

[0020] The present invention also provides a method for controlling online cell counting, comprising: Step S1: Provide the above-mentioned online cell counting device; Step S2: Connect the test container, the dilution container, and the waste liquid storage device to the online cell counting device; Step S3: Insert the microfluidic chip: Step S4: Turn on the first peristaltic pump to fill the connecting tubing with cell fluid and rinse the connecting tubing; Step S5: Select the counting mode based on the cell concentration and start online cell counting detection; Step S6: The detected cells are transported to the waste liquid storage device for collection and processing.

[0021] According to some embodiments of the present invention, the above-described online cell counting control method can detect the number of cells in real time and deliver the cells directly to the detection chip, avoiding external environmental contamination of the cells and thus affecting the detection structure. At the same time, the accuracy of the detection results is improved by rinsing the connecting pipes. The above-described control method improves the convenience for users when detecting cells.

[0022] In some embodiments of the present invention, selecting a counting mode based on the cell concentration and initiating online cell counting detection includes: Step S51: Detect or input the cell concentration, and select a counting mode based on the cell concentration; Step S52: If the low concentration counting mode is selected, control the fluid drive component to turn on the first peristaltic pump to deliver 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, while simultaneously delivering diluent into the microfluidic chip to adjust the concentration of the cells to be tested; Step S53: Turn on the laser source so that the laser passes through the focusing component and enters the cell detection channel of the microfluidic chip; Step S54: Adjust the height of the microfluidic chip so that the light passing through the cell detection channel is incident on the sensing surface of the photoelectric sensor, thereby realizing the counting of the number of cells.

[0023] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Attached Figure Description

[0024] Figure 1 This is a schematic diagram of the online cell counting device according to an embodiment of the present invention. Figure 1 ; Figure 2 This is a schematic diagram of the online cell counting device according to an embodiment of the present invention. Figure 2 ; Figure 3 This is a schematic diagram of the online cell counting device according to an embodiment of the present invention. Figure 3 ; Figure 4 This is a schematic diagram of the online cell counting device according to an embodiment of the present invention. Figure 4 ; Figure 5 This is a schematic diagram of the XY adjustment mechanism according to an embodiment of the present invention. Figure 1 ; Figure 6 This is a schematic diagram of the XY adjustment mechanism according to an embodiment of the present invention. Figure 2 ; Figure 7 This is a schematic diagram of the connection of the online cell counting device according to an embodiment of the present invention; Figure 8 The control method flow of the online cell counting device according to an embodiment of the present invention Figure 1 ; Figure 9 The control method flow of the online cell counting device according to an embodiment of the present invention Figure 2 .

[0025] Figure Labels 100. Online cell counting device; 1. Power socket; 2. Power switch; 3. Network socket; 4. Connection port; 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 component; 15. Y-axis adjustment component; 16. Pressure block; 17. Housing support; 18. Support base; 19. Pressure cap; 20. Aperture; 21. 22. USB interface; 23. Lens cover; 24. Lens bracket; 25. Through hole; 26. X-axis linear module; 27. Chip adjustment mechanism; 28. First peristaltic pump; 39. Second peristaltic pump; 30. Cell detection channel; 31. First inlet end; 32. Second inlet end; 33. Outlet end; 34. First pipeline; 35. Second pipeline; 36. Third pipeline; 37. Test container; 38. Dilution container; 49. Waste liquid storage device; 40. Fourth pipeline; 41. Fifth pipeline; 42. Base plate; 43. Bracket; 44. Outer shell; 45. Waste liquid box; 46. Control circuit board; 47. Display screen; 58. Connector; 59. First heat sink; 50. Second heat sink; 51. Third heat sink. Detailed Implementation

[0026] The online cell counting device and control method of the present invention will now be described in more detail with reference to the accompanying drawings, which illustrate preferred embodiments of the invention. It should be understood that those skilled in the art can modify the invention described herein while still achieving its advantageous effects. Therefore, the following description should be understood as being of general knowledge to those skilled in the art and is not intended to limit the invention.

[0027] In the description of this specification, terms such as "one embodiment" or "some embodiments" mean that one or more embodiments of this specification include a particular feature, structure, or characteristic described in connection with that embodiment. Therefore, the phrases "in one embodiment," "in some embodiments," "in other embodiments," "in still other embodiments," etc., appearing in different parts of this specification do not necessarily refer to the same embodiment, but rather mean "one or more, but not all, embodiments," unless otherwise specifically emphasized.

[0028] Embodiments of the present invention are described in detail below. Examples of these embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0029] In existing technologies, cell counters mostly employ offline detection modes, requiring manual transfer of samples to the detection chip for image recognition. This method suffers from poor timeliness and a high risk of contamination, especially for high-concentration cell samples where cell overlap can lead to counting errors, and sample recovery is not possible. Traditional equipment lacks automated control capabilities, making it difficult to meet the real-time monitoring needs in sterile environments, and also suffers from limitations such as large size and complex operation.

[0030] To address these challenges, researchers recognized the need for an integrated online detection system. By analyzing key aspects of sample exposure during offline operations, they proposed embedding the detection process into a closed-loop liquid path system. To tackle the challenge of detecting high-concentration cells, they considered using hydrodynamic focusing technology to optimize cell distribution. Further research revealed that dynamically adjusting the positional relationship between the optical path and the detection element can improve signal acquisition accuracy, leading to a technical concept that combines liquid path control, optical detection, and automatic adjustment.

[0031] Therefore, the present invention provides an online cell counting device 100, and the online cell counting device 100 of the present invention is described below with reference to the accompanying drawings.

[0032] The online cell counting device 100 according to an embodiment of the present invention, such as Figures 1-4 As shown, it includes: a fluid circuit unit (not labeled in the figure), including a microfluidic chip 5, a fluid driving component 6 and a connecting pipe (not labeled in the figure), the connecting pipe being connected to the microfluidic chip 5, and the fluid driving component 6 being disposed on the connecting pipe for driving fluid to flow in the microfluidic chip 5; The optical path unit (not labeled in the figure) includes a laser source 8 and a focusing component 9. The light emitted from the laser source 8 is focused into a light plate by the focusing component 9 and then incident on the microfluidic chip 5. The detection component (not labeled in the figure) is disposed on the side of the microfluidic chip 5 facing away from the focusing component 9; The control unit (not labeled in the figure) is used to control the working status of the fluid drive assembly 6, the clamp valve 10 on the connecting pipeline and the detection assembly, and to obtain cell counting results based on the electrical signals provided by the detection assembly. The detection component 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 finely adjust the position of the photoelectric sensor 12 along the X-axis and Y-axis directions, respectively.

[0033] The microfluidic chip 5 refers to a transparent substrate device with micron-level flow channels, which can be fabricated using polydimethylsiloxane material through soft photolithography, and is used to constrain the flow of a single cell layer. The fluid drive component 6 refers to a device that generates fluid power, which can be implemented using a peristaltic pump or a syringe pump to ensure continuous and stable sample delivery. The light plate refers to a sheet-like beam of light formed by a cylindrical lens, specifically implemented using a 635nm semiconductor laser in conjunction with an aspherical lens group, used for lateral illumination of the flowing cells. The XY adjustment mechanism 11 refers to a planar positioning device, which can be implemented using a precision threaded screw in conjunction with an elastic reset structure, used to compensate for alignment deviations between the optical path and the flow channels.

[0034] Specifically, the sample solution is injected into the detection area of ​​the microfluidic chip 5 through the connecting tubing. The fluid drive component 6 maintains a constant flow rate, allowing cells to pass through the light plate irradiation area in a single file. The laser beam emitted by the laser source 8 is focused by the focusing component 9 to form a light plate with a thickness of approximately 10 μm. When cells flow through this area, scattered light signals are generated. The photoelectric sensor 12 of the detection component receives the light signals and converts them into electrical pulses. 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 are precisely aligned. The control unit synchronously coordinates fluid drive, optical path stabilization, and signal processing to achieve closed-loop control.

[0035] Compared to existing technologies, this device avoids sample exposure through a closed liquid path system and improves the detection accuracy of high-concentration cells by employing dynamic optical path adjustment technology. 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 solve the signal attenuation problem caused by optical path misalignment.

[0036] In other words, this invention achieves non-destructive online detection of cell samples, eliminating the risk of contamination from manual operation. The dynamic adjustment mechanism ensures the stability of detection for samples of different concentrations, and the closed-loop liquid path design supports sample circulation and recovery. The automated control system significantly improves detection efficiency and provides a reliable solution for real-time monitoring of the cell culture process.

[0037] In some embodiments of the present invention, such as Figure 5 , Figure 6 As shown, the XY adjustment mechanism 11 includes an aperture assembly 13, an X-axis adjustment component 14, a Y-axis adjustment component 15, a pressure block 16, a housing support 17, a support base 18, and a pressure cover 19; the aperture assembly 13 includes an aperture 20, a lens (not marked in the figure), and a lens pressure cover 22, the lens having the lens support 23, the aperture 20 and the lens pressure cover 22 are both mounted on the lens support 23, and are respectively located on opposite sides of the lens; The bracket 45 and the outer shell 46 are fixed on the bracket base 18. The pressure block 16 and the pressure cover 19 are respectively disposed on opposite sides of the outer shell bracket 17 and are both fixedly connected to the outer shell bracket 17. The housing support 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 housing support 17. The X-axis adjustment member 14 and the Y-axis adjustment member 15 are both mounted on the housing support 17 and are used to finely adjust the position of the aperture assembly 13 along the X-axis and Y-axis directions, respectively.

[0038] The aperture assembly 13 refers to a combined structure consisting of the aperture 20, the lens, and the lens cap 22. Specifically, it can be implemented by stacking the annular aperture 20 made of metal or polymer material with the optical lens. The aperture 20 is used to limit the shape of the light beam, and the lens is used to focus the light. The X-axis adjustment component 14 refers to a precision displacement mechanism that moves horizontally. Specifically, it can be implemented using a threaded fine-tuning knob in conjunction with a guide rail. Rotating the knob pushes the aperture assembly 13 to move laterally. The Y-axis adjustment component 15 refers to a precision displacement mechanism that moves vertically. Specifically, it can be implemented using a spring-preloaded micrometer head structure in conjunction with a limiting slot. The longitudinal displacement is controlled by the micrometer head's advance and retraction. The pressure block 16 and the cap 19 are clamping components used to fix the outer casing support 17. Specifically, they can be implemented using aluminum alloy machined parts in conjunction with fastening bolts. The symmetrically arranged clamping structure ensures the stability of the outer casing support 17.

[0039] Specifically, the aperture assembly 13 achieves precise alignment between the aperture 20 and the lens through the lens bracket 23. During assembly, the lens is first pressed into the lens bracket 23, followed by the installation of the aperture 20 and the locking of the lens cap 22. The housing bracket 17 is fixed to the main body of the device through the bracket base 18. The pressure block 16 and the cap 19 clamp the housing bracket 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 can be operated respectively to drive the aperture assembly 13, which is inserted into the through hole 24, to perform planar two-dimensional micro-movement, so that the receiving surface of the photoelectric sensor 12 precisely coincides with the focal point of the optical path. For example, the X-axis adjustment member 14 converts rotational motion into linear displacement through a threaded pair, pushing the aperture assembly 13 to move laterally within a range of 0.1-0.5 mm; the Y-axis adjustment member 15 eliminates transmission gaps through an elastic pre-tightening mechanism, achieving precise longitudinal adjustment of 0.05-0.2 mm.

[0040] Compared with existing technologies, traditional cell counting devices often employ an integral, movable structure for their optical path adjustment mechanisms, which cannot achieve precise adjustments in independent directions within a plane. This can easily lead to mismatch between the receiving surface of the photoelectric sensor 12 and the optical path. This solution, through a separately designed independent X / Y axis adjustment mechanism, can individually correct axial deviations without disassembling the outer casing 46. This solves the problem of decreased detection sensitivity caused by accumulated assembly errors in traditional devices, while also avoiding the risk of seal damage due to repeated disassembly and reassembly.

[0041] In other words, this invention achieves precise matching between the receiving surface of the photoelectric sensor 12 and the focal point of the optical path, effectively improving the signal capture capability during high-concentration cell detection and reducing counting errors caused by optical path offset. The adjustable design of the aperture assembly 13 allows the device to adapt to microfluidic chips 5 of different sizes, enabling rapid calibration of the online detection system while maintaining a sterile operating environment.

[0042] In some embodiments of the present invention, such as 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 mounted 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 finely adjust the position of the microfluidic chip 5 along the Z-axis direction.

[0043] The X-axis linear module 25 refers to a mechanical structure that provides linear motion in the horizontal direction. Specifically, it can be implemented using a ball screw, linear guide, or linear motor to drive the chip adjustment mechanism 26 to move in the X-axis direction to adjust the horizontal position of the microfluidic chip 5.

[0044] The chip adjustment mechanism 26 refers to a positioning device including a tapered positioning pin, which can be implemented using a floating structure with elastic elements. The tapered 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 installation errors.

[0045] The tapered positioning pin refers to a positioning component with a tapered end, which can be made of stainless steel or hard alloy material. Its tapered design can realize the self-centering function and ensure the alignment accuracy between the microfluidic chip 5 and the optical path unit.

[0046] Specifically, the X-axis linear module 25 drives the chip adjustment mechanism 26 to move along the X-axis, causing the tapered positioning pin to contact the positioning hole of the microfluidic chip 5. When the tapered positioning pin is inserted into the positioning hole, its tapered surface generates a 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, preventing chip damage due to over-positioning. This achieves precise positioning of the microfluidic chip 5 in both the X and Z axes, ensuring that the cells remain perpendicularly aligned with the laser plate when flowing through the detection area.

[0047] Compared with existing technologies, traditional equipment relies on manual adjustment of chip positions, 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 conical positioning pin, eliminating human error and solving the problem of optical path misalignment caused by installation deviation of the microfluidic chip 5.

[0048] In other words, this invention can automatically perform positioning compensation of the microfluidic chip 5 during cell detection, ensuring that the laser plate is strictly perpendicular to the cell flow channel, thereby improving the stability of the signal received by the photoelectric sensor 12. This solution is particularly suitable for continuous detection scenarios of high-concentration cell samples, avoiding cell overlap counting errors caused by chip position deviation, while reducing the need for manual intervention by operators.

[0049] In some embodiments of the present invention, such as 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.

[0050] The first peristaltic pump 29 refers to a drive device that generates directional flow by periodically squeezing an elastic tube. Specifically, it can be implemented using a mechanical structure with rotatable rollers, and the liquid flow rate can be controlled by adjusting the roller speed. The second peristaltic pump 30 refers to a pump body with the same structure as the first peristaltic pump 29 but controlled independently. Specifically, it can be installed in parallel to achieve independent liquid supply through dual channels. The first peristaltic pump 29 and the second peristaltic pump 30 drive the fluid by contacting the outer wall of the tube, avoiding direct contact between the liquid and the pump body, which helps maintain a sterile environment.

[0051] Specifically, when cell concentration detection is required, the rollers of the first peristaltic pump 29 begin to rotate, continuously delivering the cell suspension in the test container 38 to the detection channel of the microfluidic chip 5 by squeezing the first tubing 35. The second peristaltic pump 30 can operate synchronously or in shifts. For example, when detecting high-concentration samples, the second peristaltic pump 30 can pump diluent from the dilution container 39 into the detection channel for online dilution. The clamp valve 10 can switch the liquid delivery path by controlling the on / off state of the third tubing 37. When sample recovery is required, the third tubing 37 is closed, allowing the liquid to flow back to the waste liquid storage device 40 via the fifth tubing 42.

[0052] Compared to existing technologies, current offline devices rely on manual operation for sample transfer and dilution, which is cumbersome and prone to contamination. This solution utilizes dual peristaltic pumps working in tandem to achieve not only automated sample delivery but also online dilution, avoiding time-sensitive errors caused by manual intervention. The contactless operation of the peristaltic pumps maintains a closed system, effectively preventing external contaminants from entering the detection system.

[0053] In other words, this invention enables automated and continuous delivery of cell suspensions, allowing for online dilution of high-concentration samples while maintaining a sterile environment. The test liquid is pumped directly into the detection area via independent tubing, avoiding the risk of sample exposure. Simultaneously, waste liquid can be selectively recycled, solving the problem of wasting precious samples. The dual-pump structure allows the dilution operation to be performed synchronously with the detection process, enabling adjustments to sample concentration without interrupting the detection process.

[0054] It should be noted that the online cell counting device involved in this invention, by incorporating the waste liquid storage device 40, can effectively collect waste liquid during the detection process, avoiding environmental pollution. The waste liquid storage device 40 is connected to the detection device and the first peristaltic pump 29 via the connecting pipe, allowing waste liquid to flow smoothly from the detection device into the waste liquid storage device 40. This makes the entire detection process cleaner and safer, preventing the indiscriminate discharge of waste liquid.

[0055] The installation position of the waste liquid storage device 40 can be flexibly designed. For example, it can be fixed to one side of the housing, that is, a waste liquid box 47 can be set on the housing. Figure 1As shown, the waste liquid storage device 40 is placed inside the waste liquid box 47. The waste liquid storage device 40 can be configured as a waste liquid bag, and its specific position 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 testing, thus improving testing efficiency. The waste liquid storage device 40 can adopt a sealed structure to prevent the evaporation and leakage of waste liquid, further ensuring the cleanliness and safety of the testing environment.

[0056] By incorporating the waste liquid storage device 40, the online cell counting device of the present invention can effectively collect and treat waste liquid during the detection process, avoiding environmental pollution and ensuring the cleanliness and safety of the detection process. Furthermore, the waste liquid storage device 40 is flexibly designed and can be adjusted according to actual needs, improving the applicability and practicality of the device. Compared with existing technologies, the technical solution of the present invention has significant advantages in waste liquid treatment and can better meet actual detection requirements. In some embodiments of the present invention, such as 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 communicating with 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 test container 38, and the other end of the first pipeline 35 is connected to one end of the third pipeline 37 through the connector 50. The other end of the third pipeline 37 is connected to the first inlet end 32 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 33 of the microfluidic chip 5; the first peristaltic pump 29 is disposed on the first pipeline 35, the second peristaltic pump 30 is disposed on the second pipeline 36, and the clamp valve 10 is disposed on the third pipeline 37.

[0057] The cell detection channel 31 refers to a channel for cell flow and detection. Specifically, it can be implemented using a microchannel structure with a rectangular or circular cross-section, with a width or diameter ranging from 50 to 200 micrometers, which can constrain the single-line flow of cells.

[0058] The first inlet end 32 and the second inlet end 33 are interfaces used to introduce the sample to be tested and the diluent, respectively. Specifically, they can be implemented using a tapered tapered structure, which helps to reduce turbulence during fluid mixing.

[0059] The connector 50 refers to a component used for connecting pipelines, which can be implemented using a tee connector or a quick-connect connector to facilitate switching and sealing between multiple pipelines.

[0060] Specifically, the sample to be tested is driven by the first peristaltic pump 29 through the first conduit 35, and the diluent is driven by the second peristaltic pump 30 through the second conduit 36. The two pumps converge at the connector 50 and then enter the first inlet 32 ​​of the microfluidic chip 5 through the third conduit 37. The clamp valve 10 regulates the flow rate of the mixture by controlling the opening and closing of the third conduit 37, while the second inlet 33 directly receives the diluent for secondary dilution. By adjusting the rotational speed ratio of the two peristaltic pumps, the mixing ratio of the sample and the diluent can be precisely controlled, allowing the high-concentration cell solution to undergo gradient dilution before flowing through the detection channel, avoiding counting errors caused by cell overlap.

[0061] Compared to existing technologies, which require multiple manual dilutions and chip replacements for offline detection, this solution achieves online automated dilution through multi-channel coordinated control, significantly reducing operation time while maintaining a sterile environment. Furthermore, in existing technologies, the mixing ratio of diluent and sample depends on the precision of manual pipetting, while this solution achieves precise, adjustable mixing through an independently controlled peristaltic pump.

[0062] In other words, this invention solves the problem of repeated manual dilution required for high-concentration cell detection, avoiding the risk of sample exposure and contamination. Furthermore, the multi-stage dilution channel design improves detection accuracy. The dynamic adjustment function of the mixing ratio can also adapt to the detection needs of different cell concentrations, enhancing the versatility of the device.

[0063] In some embodiments of the present invention, such as Figure 7 As shown, it also includes a waste liquid storage device 40. The connecting pipelines also include a fourth pipeline 41 and a fifth pipeline 42. One end of the fourth pipeline 41 and the fifth pipeline 42 are connected to the waste liquid storage device 40. 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 connector 50. The pinch valve 10 is a two-way pinch valve 10. The opening and closing of the third pipeline 37 and the fifth pipeline 42 are both controlled by the pinch valve 10.

[0064] The waste liquid storage device 40 is a container used to collect waste liquid generated during the testing process. It can be implemented as a sealed storage tank to prevent waste liquid leakage and environmental pollution. The fourth pipeline 41 is a transmission channel connecting the outlet end 34 of the microfluidic chip 5 to the waste liquid storage device 40. It can be implemented as a flexible silicone tube to ensure smooth discharge of waste liquid. The fifth pipeline 42 is an auxiliary channel connecting the waste liquid storage device 40 to the first pipeline 35 and the third pipeline 37. It can be implemented as a tee connector to achieve multi-directional fluid communication. The two-way pinch valve 10 is a valve that can simultaneously control the opening and closing of two pipelines. It can be implemented as an electromagnetically driven pinch valve 10, with synchronous opening and closing actions achieved through a control circuit.

[0065] Specifically, during cell detection, when sample retrieval is required, the pinch valve 10 simultaneously closes the third pipeline 37 and opens the fifth pipeline 42, allowing the original sample in the first pipeline 35 to flow directly back to the waste liquid storage device 40 for temporary storage via the fifth pipeline 42. When waste liquid discharge is required, the pinch valve 10 closes the fifth pipeline 42 and opens the third pipeline 37, allowing the detected waste liquid to be discharged into the waste liquid storage device 40 via the fourth pipeline 41. This dual-pipeline design creates independent channels for sample retrieval and waste liquid discharge, enabling rapid switching between the two operating modes using a single valve.

[0066] Compared to existing technologies, traditional cell counting devices typically employ open waste tanks with only a single discharge channel, failing to achieve sample recovery and posing a risk of cross-contamination. This solution, by adding the fifth pipeline 42 and the two-way pinch valve 10, achieves both waste discharge and sample recovery functions while maintaining the compactness of the piping system. In particular, the synchronous control characteristic of the two-way pinch valve 10 avoids the complexity of requiring multiple valves to operate in coordination as in traditional solutions.

[0067] In other words, this invention effectively solves the technical problem of the inability to recover rare cell samples. Unused original samples can be safely and temporarily stored during the testing process, avoiding sample waste caused by traditional offline testing. The dual-pipe diversion 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 application of the two-way pinch valve 10 further simplifies the equipment operation process, giving the online cell counting device 100 a higher degree of automation.

[0068] In some embodiments of the present invention, a device body (not shown in the figure) is also included, such as Figures 1-4As shown, the main body of the device includes a base plate 44, a bracket 45, and a housing 46. The bracket 45 is mounted on the base plate 44, the detection component is mounted on the bracket 45, the bottom surface of the housing 46 is fixed to the base plate 44, and the fluid drive component 6 and the switch are both mounted on the top surface of the housing 46.

[0069] The base plate 44 refers to the support structure that supports the internal components of the main body of the device. Specifically, it can be made of metal sheet or composite material plate, and is used to provide rigid support for the bracket 45 and the outer shell 46.

[0070] The bracket 45 refers to a support frame that is vertically installed on the base plate 44. Specifically, it can be made of aluminum alloy profile or injection molded structure, and is used to fix the detection component and maintain its spatial position stability.

[0071] The outer shell 46 refers to the protective shell covering the internal structure of the main body of the device. Specifically, the outer shell 46 can be made by injection molding in a split manner or by splicing sheet metal to form the outer shell 46, which is used to isolate the external environment from interference with the internal components.

[0072] The switching element refers to the actuator that controls the opening and closing of the pipeline, which can be implemented by a solenoid valve, the pinch valve 10, or a rotary valve, and is used to adjust the liquid flow path.

[0073] Specifically, the base plate 44 is rigidly connected to the bracket 45 by bolts or welding. The detection component is fixed to a preset mounting position on the bracket 45 by screws or clips. The housing 46 is sealed to the base plate 44 by edge grooves. The fluid drive component 6 and the switch are mounted in the reserved mounting area on the top surface of the housing 46 via guide rails or positioning pins. Their control cables pass through the cable holes on the side wall of the housing 46 and connect to the internal circuitry. Thus, the main body of the device forms a layered layout structure. The base plate 44 provides mechanical support, the bracket 45 enables precise positioning of the detection component, and the housing 46 provides physical protection and integrates the external operating interface.

[0074] Compared to existing technologies, current cell counting devices typically employ a split structure, with the fluid drive component and detection component installed separately, resulting in bulky equipment and difficult maintenance. This solution integrates the main body of the device, centrally arranging the fluid drive component 6 and the switching components on the top surface of the housing 46. This facilitates quick inspection and replacement of components by operators, while the rigid connection between the bracket 45 and the base plate 44 ensures the positional accuracy of the detection component.

[0075] In other words, the present invention achieves a compact and modular main structure for the device, solving the problems of large size and inconvenient maintenance of existing equipment. The rigid connection between the base plate 44 and the bracket 45 ensures the positioning stability of the detection component, the sealed design of the outer shell 46 reduces the risk of external contamination, and the optimized top surface layout of the fluid drive component 6 and the switch optimizes the ease of operation, thereby supporting the stable operation of the online cell counting device 100 in a continuous production environment.

[0076] In some embodiments of the present invention, such as Figure 3 , Figure 4 As shown, the laser 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 a chip slot, the chip slot is located at the end of the focusing component 9 away from the laser source 8, the microfluidic chip 5 is installed in the chip slot, the liquid circuit unit connects the dilution container 39, the test container 38 and the waste liquid storage device 40, and the laser source 8, the focusing component 9, the microfluidic chip 5, the aperture component 13 and the photoelectric sensor 12 are located on the same straight line.

[0077] The laser source 8 refers to a device for generating the excitation beam required for detection. Specifically, it can be implemented using a semiconductor laser, with a wavelength range of, for example, 405-650nm, to excite the cells flowing through the microfluidic chip 5 to generate light signals.

[0078] The X-axis linear module 25 refers to the mechanical structure that drives the microfluidic chip 5 to perform horizontal displacement. Specifically, it can be implemented 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.

[0079] The chip slot refers to the positioning structure that fixes the microfluidic chip 5. Specifically, it can be implemented by using an aluminum alloy groove with elastic buckles. The inner wall of the groove can be provided with a tapered positioning pin that cooperates with the positioning hole of the chip to achieve precise positioning of the chip in the vertical direction.

[0080] The connection between the liquid circuit unit and the dilution container 39, the test container 38, and the waste liquid storage device 40 refers to the pipeline connection method for constructing a closed fluid circulation system. Specifically, it can be achieved by using a combination of medical-grade silicone tubing and Luer locking connectors to realize automatic sample dilution, transportation, and waste liquid recovery.

[0081] Specifically, the excitation beam emitted by the laser source 8 is focused by 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 liquid containing cells flows through the detection area, the cells trigger scattered light and fluorescence signals as they pass through the light plate area. These signals are captured by the photoelectric sensor 12 and converted into electrical signals. The X-axis linear module 25 drives the chip slot to move horizontally, aligning different detection areas sequentially with the optical path system. The liquid path unit uses a peristaltic pump to control the automatic dilution and transport of samples, and the waste liquid storage device 40 is connected via an airtight pipeline to ensure sterility during operation. The design of all optical elements arranged along the same straight line reduces optical path transmission loss to below 5%.

[0082] Compared to existing technologies, traditional cell counters require manual chip placement and cannot achieve continuous detection. This solution, however, achieves online continuous sample detection through an integrated liquid path system and automatic positioning mechanism. Existing devices often employ discrete optical systems, resulting in bulky designs. This solution integrates optical components onto a single support 45 using 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 liquid path unit, can dynamically adjust the sample concentration to a suitable detection range of 1E5-1E7 cells / ml.

[0083] In other words, this invention achieves fully automated cell counting, avoiding the contamination risks associated with manual operation. The closed-loop liquid system maintains sample sterility, making it particularly suitable for bioreactor scenarios requiring continuous monitoring. The collinear layout of the optical components effectively improves signal acquisition efficiency, enabling detection sensitivity at the single-cell level. The modular design of the X-axis linear module 25 and the chip slot structure allows the device to adapt to detection chips of different specifications, expanding its applicability.

[0084] In some embodiments of the present invention, such as 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, and 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 clamp valve 10, and the display screen 49.

[0085] The control circuit board 48 refers to an integrated circuit board used to receive and process electrical signals and output control commands. Specifically, it can be implemented using a multi-layer PCB board integrating a microprocessor and drive circuit, used to coordinate the timing of actions of various execution components. The display screen 49 refers to a visual output device for human-computer interaction, specifically a touch-screen LCD screen, used to display cell count data and system status parameters in real time. The top surface of the outer shell 46 refers to the planar area of ​​the shell covering the upper part of the device body, specifically made of metal or engineering plastic, used to support the display screen 49 and form a sealed structure. The bracket 45 refers to a rigid frame structure supporting the detection components, specifically welded from aluminum alloy profiles, used to fix the control circuit board 48 and maintain its spatial stability.

[0086] Specifically, the control circuit board 48 establishes a signal transmission channel with the photoelectric sensor 12 via wires, receiving pulse signals generated in real time as cells pass through the microfluidic chip 5. The stepper motor drive signal for the moving component is generated by the control circuit board 48, and chip positioning is achieved by adjusting the displacement of the X-axis linear module 25. The peristaltic pump speed command for the fluid drive component 6 is output by the control circuit board 48, controlling the liquid flow rate by changing the pulse frequency. The opening and closing state of the clamp valve 10 is switched by high and low level signals sent by the control circuit board 48, achieving precise control of pipeline on / off. The display screen 49 is connected to the control circuit board 48 via a ribbon cable, dynamically refreshing the processed cell concentration data in chart form. The tilt angle of the top surface of the housing 46 is calculated ergonomically, so that the display screen 49 faces the operator at a 15-degree angle. The bracket 45 has internal wiring channels for orderly arranging the connecting cables between the control circuit board 48 and each actuator.

[0087] In some specific embodiments, the control circuit board 48 can be divided into independent functional areas. For example, the signal acquisition area may be equipped with an AD conversion chip, the motion control area may be equipped with a stepper motor driver chip, and the fluid control area may integrate a MOSFET switching circuit. The display screen 49 can be installed using a magnetic quick-release structure for easy and rapid replacement during maintenance. The bracket 45 is preferably fixed to the control circuit board 48 using insulated nylon posts and screws to avoid the risk of short circuits.

[0088] Compared to existing technologies, the control modules of traditional cell counters are typically distributed, resulting in bulky devices and difficulties in heat dissipation. This solution integrates the control circuit board 48 inside the bracket 45, significantly shortening the signal transmission distance between the sensor and the processor and effectively reducing electromagnetic interference. The integrated design of the display screen 49 and the top surface of the housing 46 replaces the traditional external display cable, making the user interface layout more compact and rational. The direct connection between the control circuit board 48 and the actuator eliminates the traditional relay control mode, improving system response speed and reliability.

[0089] In other words, this invention achieves centralized processing of control commands for multiple subsystems, ensuring precise synchronization of actions such as fluid drive, optical detection, and mechanical positioning. The real-time data feedback from the display screen 49 allows the operator to intuitively monitor changes in cell concentration, avoiding detection errors caused by information lag. The integrated installation of the control circuit board 48 and the bracket 45 optimizes the utilization of internal space, providing ample layout space for the heat dissipation components, thereby meeting the stability requirements for long-term continuous operation.

[0090] In some embodiments of the present invention, such as Figure 3 , Figure 4 As shown, it also includes a heat dissipation assembly, which (not marked in the figure) includes a first heat dissipation component 51, a second heat dissipation component 52 and a third heat dissipation component 53. The first heat dissipation component 51 is mounted on the laser light source 8, and the second heat dissipation component 52 and the third heat dissipation component 53 are both mounted on the base plate 44. The second heat dissipation component 52 is located in the middle of the base plate 44, and the third heat dissipation component 53 is located at the edge of the base plate 44.

[0091] The first heat sink 51 refers to the heat dissipation component that is in direct contact with the laser source 8, and can be implemented using a metal heat sink or a miniature fan, for quickly dissipating the heat generated by the laser source 8. The second heat sink 52 refers to the heat dissipation structure located in the middle of the base plate 44, and can be implemented using heat sink fins or heat pipes, for balancing the concentrated heat generated during device operation. The third heat sink 53 refers to the auxiliary heat dissipation component located at the edge of the base plate 44, and can be implemented using heat dissipation grilles or heat dissipation holes, for enhancing airflow efficiency in the edge area of ​​the device.

[0092] Specifically, the laser source 8 generates a large amount of heat during prolonged operation. The first heat sink 51 conducts this heat to the external environment through physical contact. The center of the base plate 44, which supports the fluid drive assembly 6 and the control unit, is prone to heat accumulation. The second heat sink 52 accelerates heat dissipation in this area by increasing the heat dissipation area. The edges of the base plate 44, being close to the enclosed area of ​​the outer shell 46, are prone to heat dissipation dead zones. The third heat sink 53 improves the heat dissipation efficiency of the edge area by optimizing the airflow path. These three sets of heat sinks are strategically positioned to address different heat source distribution characteristics, forming a collaborative heat dissipation mechanism.

[0093] Compared to existing technologies, traditional cell counting devices typically only have a single heat dissipation structure for key components, which cannot cope with the combined heat load generated by the simultaneous operation of the high-power laser source 8, the fluid drive component 6, and the control unit. This solution, by implementing a zoned heat dissipation structure, can provide targeted heat dissipation for the laser source 8, the core area, and the edge areas of the device, effectively avoiding problems such as optical path deviation, fluid parameter fluctuations, and performance degradation of electronic components caused by excessively high local temperatures.

[0094] In other words, this invention solves the problems of decreased detection accuracy and insufficient equipment stability caused by uneven heat dissipation in existing cell counting devices. The zoned heat dissipation design can maintain the stability of the laser wavelength in the optical path unit, avoid fluctuations in fluid drive velocity caused by temperature changes, and extend the service life of the electronic components of the control unit, ensuring the data reliability of the cell counting process.

[0095] Furthermore, such as Figure 2 As shown, multiple ventilation openings 7 can also be provided on the outer casing to further accelerate the air circulation inside the casing, thereby improving the working efficiency of the heat dissipation components and enhancing the heat dissipation capacity.

[0096] In some embodiments of the present invention, such as Figure 2 As shown, it also includes a power socket 1, which is mounted on the bracket. The outer casing has a first hole corresponding 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.

[0097] The power socket 1 ensures a stable power supply for the device. By mounting the power socket 1 on the bracket and providing a corresponding first hole on the outer casing, power connection is made more convenient and effectively avoids exposed wires, improving the overall aesthetics and safety of the device. Furthermore, a power switch and an indicator light can be installed on the bracket. These are electrically connected to the power socket 1, passing through the outer casing and exposed to the external environment. The power switch further controls the power supply to the device, improving circuit safety; the indicator light indicates the circuit status, enhancing ease of use.

[0098] The power connector 1 can be a standard power socket or a USB interface 21, or other interfaces capable of connecting to a power source. The power connector 1 is installed in a suitable position on the bracket and exposes itself to the external environment through a first hole in the housing. The first hole can be sealed to prevent external environmental influences on the circuitry. The power connector 1 is connected to the detection component, the control circuit board, and the laser source 8 via wires, ensuring stable operation of these components. As a preferred embodiment, the power connector 1 can be a quick-connect design, allowing users to quickly connect and disconnect the power supply.

[0099] Through the above design, the online cell counting device of the present invention offers greater convenience in power connection, improving the user experience and safety. Compared with existing technologies, the solution of the present invention effectively solves the problem of inconvenient power connection, ensures stable power supply to the device, and enhances overall reliability and aesthetics.

[0100] Furthermore, such as Figure 1 As shown, the outer casing is also equipped with a power switch 2, which is electrically connected to the power socket 1 to realize the power control of the device and further improve the safety and stability of the device's power supply.

[0101] In some embodiments of the present invention, such as Figure 2 As shown, it also includes a network port 3, which is mounted on the bracket. The housing has a second hole (not marked in the figure) corresponding to the network port 3. The network port 3 is electrically connected to the detection component, the control circuit board and the fluid drive component 6.

[0102] The network port 3 enables the online cell counting device to transmit data and be remotely controlled via a network interface. Specifically, by connecting to a network, the network port 3 allows the detection component, the control circuit board, and the drive component to communicate with an external computer or server, thereby achieving real-time data transmission and remote monitoring. Thus, users can remotely access and control the online cell counting device via the network, obtain real-time detection data, and perform corresponding operations and adjustments.

[0103] In a preferred embodiment, the network port 3 can adopt a standard Ethernet interface, such as an RJ45 interface, to ensure compatibility with existing network devices. Furthermore, the network port 3 can be protected by a shielding cover to prevent external interference and physical damage.

[0104] By adding the network port 3 to the device, the present invention effectively solves the problems of manual operation and inconvenient data transmission in existing offline cell counters. Compared with the prior art, the advantages of the present invention are that it realizes online real-time data transmission and remote control, improves detection efficiency and data reliability, and reduces the risk of human interference and contamination.

[0105] Furthermore, such as 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 cable, thereby enabling the online cell counting device to be controlled by an external device, or to be analyzed and counted by an external device. Furthermore, the connection port 4 can also be connected to other external devices to achieve more functions, which will not be elaborated here.

[0106] In addition, such as Figure 1 As shown, the online cell counting device also includes a USB interface 21, which is used to connect to external devices. Specifically, by setting the USB interface 21, an external power supply can be connected instead of the power socket 1. By setting a different socket type than 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 external devices, enabling the online cell counting device to connect to multiple external devices simultaneously. Furthermore, by setting an interface of a different type than the connection port 4, the versatility of the online cell counting device is improved.

[0107] The present invention also provides a control method for online cell counting, the control method for online cell counting including the online cell counting device 100 described above, such as... Figure 8 As shown, the control methods include: Step S1: Provide the above-mentioned online cell counting device 100; Step S2: Connect the test container 38, the dilution container 39, and the waste liquid storage device 40 to the online cell counting device 100; Step S3: Insert the microfluidic chip 5: Step S4: Turn on the first peristaltic pump 29 to fill the connecting tubing with cell fluid and rinse the connecting tubing; Step S5: Select the counting mode based on the cell concentration and start online cell counting detection; Step S6: The detected cells are transported to the waste liquid storage device 40 for collection and processing.

[0108] Specifically, the power supply is connected to the power port, and a network cable is connected to the network port 3, enabling the device to communicate with external devices via the network. The microfluidic chip 5 is inserted into the chip mounting component 130. The microfluidic chip 5 is connected to one port of the switch component via the connecting pipe, thereby connecting to the test container 38. The first peristaltic pump 29 is connected between the switch component and the test container 38 via 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 via the connecting pipe. The microfluidic chip 5 is directly connected to the waste liquid storage device 40 via the connecting pipe. The waste liquid storage device 40 is directly connected to the test container 38 via the switch component.

[0109] In other words, the online cell counting device 100 is first connected to a power source and network, the microfluidic chip 5 is installed, and the test container 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 controls the switch 6 to connect the waste liquid storage device 40 and the test container 38, controlling the first peristaltic pump 29 to transport the test cell solution from the test container 38 to the waste liquid storage device 40 to rinse the connecting tubing, preventing residual substances in the connecting tubing from affecting the experimental results and improving the accuracy of the detection. Further, the control circuit board 48 controls the switch 6 to turn off the waste liquid storage device 40. The liquid storage device 40 is connected to the test container 38, which in turn connects the test container 38 and the microfluidic chip 5. The first peristaltic pump 29 is controlled to deliver the test cell fluid to the microfluidic chip 5. Then, the control circuit board 48 is operated to turn on the laser source 8, and the focusing component 9, the XY adjustment mechanism 11, and the microfluidic chip 5 are adjusted to a suitable position so that the laser passes through the focusing channel of the microfluidic chip 5. The scattered 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 for cell counting and cell size analysis. After the cell counting detection is completed, the first peristaltic pump 29 is controlled to deliver the detected cell fluid to the waste liquid storage device 40 for waste liquid collection and treatment.

[0110] In some embodiments of the present invention, such as Figure 9 As shown, the counting mode is selected according to the cell concentration, and online cell counting detection is started, including: Step S51: Detect or input cell concentration, and select the counting mode according to the cell concentration; Step S52: If the low concentration counting mode is selected, the fluid drive component 6 is controlled to turn on the first peristaltic pump 29 to deliver the cells to be tested to the microfluidic chip 5; 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 diluent is delivered into the microfluidic chip 5 to adjust the concentration of the cells to be tested. Step S53: Turn on the laser source 8 so that the laser passes through the focusing component 9 and enters the cell detection channel 31 of the microfluidic chip 5; Step S54: Adjust the height of the microfluidic chip 5 so that the light passing through the cell detection channel 31 is incident on the sensing surface of the photoelectric sensor 12, thereby realizing the counting of the number of cells.

[0111] Specifically, the normal concentration detection range of the microfluidic chip 5 is <5E7 / ml. When the cell fluid concentration in the test container 38 meets the requirement of <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 diluent to the microfluidic chip 5 to dilute the concentration of the test cell fluid, so that the concentration of the test cell fluid meets the requirement of <5E7 / ml, thereby making the counting of the microfluidic chip 5 accurate.

[0112] It should be understood that the specific embodiments described above are merely illustrative or explanatory of the principles of the invention and do not constitute a limitation thereof. Therefore, any modifications, equivalent substitutions, improvements, etc., made without departing from the spirit and scope of the invention should be included within the protection scope of the invention. Furthermore, the appended claims are intended to cover all variations and modifications falling within the scope and boundaries of the appended claims, or equivalent forms of such scope and boundaries.

Claims

1. An online cell counting device, characterized in that, include: The fluid circuit unit includes a microfluidic chip, a fluid driving component, and a connecting pipe. The connecting pipe is connected to the microfluidic chip, and the fluid driving component is disposed on the connecting pipe for driving fluid to flow in the microfluidic chip. The optical path unit includes a laser source and a focusing component. The emitted light from the laser source is focused into a light plate by the focusing component and then incident on the microfluidic chip. A detection component is disposed on the side of the microfluidic chip opposite to the focusing component; The control unit is used to control the working status of the fluid drive assembly, the clamp valve on the connecting pipeline and the detection assembly, and to obtain cell counting results based on the electrical signal provided by the detection assembly. The detection component includes an XY adjustment mechanism and a photoelectric sensor mounted on the XY adjustment mechanism. The XY adjustment mechanism is used to finely adjust the position of the photoelectric sensor along the X-axis and Y-axis directions, respectively. The XY adjustment mechanism includes an aperture assembly, an X-axis adjustment component, a Y-axis adjustment component, a pressure block, a housing support, a support base, and a pressure cap. The aperture assembly includes an aperture, a lens, and a lens pressure cap. The lens has a lens support, and the aperture and the lens pressure cap are both mounted on the lens support and located on opposite sides of the lens. The housing support is fixed to the support base, and the pressure block and the pressure cap are respectively located on opposite sides of the housing support and are fixedly connected to the housing support. The housing support has a through hole, and one end of the aperture assembly is inserted into the through hole of the housing support. The X-axis adjustment component and the Y-axis adjustment component are both mounted on the housing support and are used to finely adjust the position of the aperture assembly along the X-axis and Y-axis directions, respectively. The online cell counting device also includes an X-axis linear module and a chip adjustment mechanism; the chip adjustment mechanism is mounted on the X-axis linear module and can move back and forth along the X-axis; the tapered positioning pin of the chip adjustment mechanism cooperates with the positioning hole on the microfluidic chip to finely adjust the position of the microfluidic chip along the Z-axis.

2. 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 liquid from the test container into the microfluidic chip; the second peristaltic pump is used to pump liquid from the dilution container into the microfluidic chip.

3. The online cell counting device according to claim 2, characterized in that, The microfluidic chip has a cell detection channel and a first inlet, a second inlet, and an outlet communicating with 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 test container, the other end of the first pipeline is connected to one end of the third pipeline via a connector, the other end of the third pipeline is connected to the first inlet 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 of the microfluidic chip; a first peristaltic pump is disposed on the first pipeline, a second peristaltic pump is disposed on the second pipeline, and the clamp valve is disposed on the third pipeline.

4. The online cell counting device according to claim 3, characterized in that, It also includes a waste liquid storage device, and the connecting pipeline further includes a fourth pipeline and a fifth pipeline. One end of each of the fourth and fifth pipelines 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 connector. 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.

5. The online cell counting device according to claim 4, characterized in that, It also includes a device body, which includes a base plate, a bracket, and a housing. The bracket is mounted on the base plate, the detection component is mounted on the bracket, the bottom surface of the housing is fixed to the base plate, and the fluid drive component and the switch are both mounted on the top surface of the housing.

6. The online cell counting device according to claim 5, characterized in that, The laser source is mounted on the bracket, the X-axis linear module is mounted on the base plate below the bracket, the bracket is provided with a chip slot, the chip slot is located at the end of the focusing component away from the laser source, the microfluidic chip is mounted in the chip slot, the liquid circuit unit connects the dilution container, the test container and the waste liquid storage device, and the laser source, the focusing component, the microfluidic chip, the aperture component and the photoelectric sensor are located on the same straight line.

7. The online cell counting device according to claim 5, 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, and 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 clamp valve, and the display screen. The stepper motor drive signal for the moving component is generated by the control circuit board, and chip positioning is achieved by adjusting the displacement of the X-axis linear module.

8. The online cell counting device according to claim 5, characterized in that, It also includes a heat dissipation assembly, which includes a first heat dissipation component, a second heat dissipation component, and a third heat dissipation component. The first heat dissipation component is mounted on the laser light source, and the second and third heat dissipation components are both mounted on the base plate. 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.

9. A method for controlling online cell counting, characterized in that, include: Step S1: Provide an online cell counting device as described in any one of claims 2-8; Step S2: Connect the test container, the dilution container, and the waste liquid storage device to the online cell counting device; Step S3: Insert the microfluidic chip: Step S4: Turn on the first peristaltic pump to fill the connecting tubing with cell fluid and rinse the connecting tubing; Step S5: Select the counting mode based on the cell concentration and start online cell counting detection; Step S6: The detected cells are transported to the waste liquid storage device for collection and processing.

10. The online cell counting control method according to claim 9, characterized in that, Select the counting mode based on the cell concentration and begin online cell counting detection, including: Step S51: Detect or input the cell concentration, and select a counting mode based on the cell concentration; Step S52: If the low concentration counting mode is selected, control the fluid drive component to turn on the first peristaltic pump to deliver 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, while simultaneously delivering diluent into the microfluidic chip to adjust the concentration of the cells to be tested; Step S53: Turn on the laser source so that the laser passes through the focusing component and enters the cell detection channel of the microfluidic chip; Step S54: Adjust the height of the microfluidic chip so that the light passing through the cell detection channel is incident on the sensing surface of the photoelectric sensor, thereby realizing the counting of the number of cells.

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