CN118389242B

CN118389242B - Cell screening device

Info

Publication number: CN118389242B
Application number: CN202410618497.7A
Authority: CN
Inventors: 杨琴
Original assignee: Guangzhou Yitaitong Biotechnology Co ltd
Current assignee: Guangzhou Yitaitong Biotechnology Co ltd
Priority date: 2024-05-17
Filing date: 2024-05-17
Publication date: 2024-11-01
Anticipated expiration: 2044-05-17
Also published as: CN118389242A

Abstract

The invention discloses a cell screening device, which relates to the technical field of cell screening, and comprises a sheath fluid inlet pipe, a cell suspension pipe and a flow chamber which are arranged on a body, and further comprises the following components: the buffer part comprises a buffer tube fixedly communicated with the sheath liquid inlet tube, and one end of the buffer tube, which is far away from the sheath liquid inlet tube, is fixedly communicated with a sheath liquid outlet tube; the bubble removing part comprises a guide pipe and a thin pipe which are fixedly communicated with the buffer pipe, and a venturi is fixedly communicated between the guide pipe and the thin pipe; and a pH measuring section including a pH probe. The advantages are that: according to the invention, stirring, bubble removal and pH value measurement treatment of the sheath fluid can be automatically completed by utilizing the change of the sheath fluid flow in the sheath fluid flow process, so that the uniform mixing of the sheath fluid components is ensured, the influence on the cell screening result caused by the mixing of bubbles in the sheath fluid into a flow chamber is avoided, the cell screening effect is ensured, the overall automation degree is high, and the structure is simple.

Description

Cell screening device

Technical Field

The invention relates to the technical field of cell screening, in particular to a cell screening device.

Background

Cell screening is a process of separating different types of cells or cell subsets from a mixture, and pure cell groups can be obtained through cell screening for further research or application, and currently common cell screening devices include a flow cytometer, a cell sorting system, a magnetic bead sorting system, a microfluidic chip and the like, wherein the flow cytometer is commonly used.

The flow cytometer works on the principle that cells are suspended in a liquid and coated by a sheath liquid, so that the cells sequentially pass through a laser beam in sequence, and when single cells pass through the laser beam, each cell can be measured and analyzed according to the characteristics of the size, shape, surface marks, internal fluorescence and the like of the cell, so that analysis and sorting of the single cells are performed.

In the use process of the flow cytometer, the deviation of cells caused by the air bubbles entering the flow chamber needs to be avoided, the orthogonal state of the laser beam and the cells is influenced, and the air bubbles are generally mixed into the flow chamber through sheath liquid at present, so that the removal of the air bubbles in the sheath liquid is an important step in the process of cell screening, but the conventional flow cytometer generally does not have the function of removing the air bubbles in the sheath liquid, so that the deviation exists in the detection result in the screening process, and therefore, a cell screening device needs to be designed.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a cell screening device, which solves the problem that bubbles in sheath fluid provided in the background art affect the detection result.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

A cell screening device comprising a sheath fluid inlet tube, a cell suspension tube and a flow chamber which are arranged on a body, and further comprising:

The buffer part comprises a buffer tube fixedly communicated with the sheath liquid inlet tube, one end of the buffer tube, far away from the sheath liquid inlet tube, is fixedly communicated with a sheath liquid outlet tube, the other end of the sheath liquid outlet tube is fixedly communicated with a nozzle ultrasonic oscillator, the nozzle ultrasonic oscillator is fixedly communicated with the flow chamber, and a baffle is fixedly arranged in the buffer tube;

The device comprises a buffer tube, a bubble removing part, a stirring assembly and a gas exhaust assembly, wherein the bubble removing part comprises a guide tube and a thin tube which are fixedly communicated with the buffer tube;

The pH measuring part comprises a pH probe, an electric push rod is fixedly installed on the buffer tube, a fixed block is fixedly installed between the output end of the electric push rod and the pH probe, a control component matched with the electric push rod is installed on the buffer tube, and the control component is matched with the stirring component.

Further, the sheath liquid inlet pipe and the catheter are both positioned on the left side of the partition board, the thin pipe and the sheath liquid outlet pipe are both positioned on the right side of the partition board, and the diameter of the catheter is larger than that of the thin pipe.

Further, the stirring assembly comprises a rotating rod which is connected to the venturi tube in a sealing and rotating mode, and a plurality of impellers are fixedly arranged at one end of the rotating rod, located in the venturi tube.

Further, the exhaust assembly comprises a rotating shaft which is connected to the unidirectional exhaust pipe in a sealing and rotating mode, and a plurality of exhaust fan blades are fixedly arranged at one end of the rotating shaft, located in the unidirectional exhaust pipe.

Further, a belt transmission structure is arranged between the rotating rod and one end of the rotating shaft, which is positioned outside the venturi tube and the unidirectional exhaust tube.

Further, the belt transmission structure comprises a belt wheel I, a belt wheel II and a belt, wherein the belt wheel I is fixedly arranged on the rotating rod, the belt wheel II is fixedly arranged on the rotating shaft, the belt is sleeved between the belt wheel I and the belt wheel II, and the diameter of the belt wheel I is larger than that of the belt wheel II.

Further, the control assembly comprises a supporting rod fixedly arranged on the buffer tube, a fixed rack is slidably arranged on the supporting rod, a fixed gear ring is fixedly arranged at one end of the rotating rod, which is far away from the first belt wheel, and is meshed with the fixed rack, a protruding block is fixedly arranged on the supporting rod, a touch switch electrically connected with the electric push rod is fixedly arranged at the bottom of the protruding block, and a touch block corresponding to the position of the touch switch is fixedly arranged at the top of the fixed rack.

Further, a T-shaped block is fixedly arranged on the side wall of the fixed rack, a chute matched with the T-shaped block is arranged on the side wall of the supporting rod, and a spring is arranged between the chute and the T-shaped block.

Further, the maximum distance between the touch switch and the touch block is equal to the length of the fixed rack.

Further, a through hole is formed in the buffer tube, and a sealing ring in sealing sliding connection with the pH probe is fixedly arranged in the through hole.

Compared with the prior art, the invention has the advantages that:

1: the cell screening device has the advantage of removing bubbles in the sheath liquid, avoids the influence of the mixing of the bubbles into the flowing chamber on the cell screening result, and can perform bubble removal treatment on the sheath liquid entering the buffer tube through the cooperation of the buffer part and the bubble removal part.

2: The cell screening device has the advantage of high automation degree, can automatically complete the degassing treatment of the sheath fluid in the flowing process of the sheath fluid, and particularly provides stable driving force for stirring and degassing of the sheath fluid through the matching of the stirring assembly and the exhaust assembly without additional control.

3: The cell screening device has the advantage of automatically measuring the pH value of the sheath liquid, ensures that the used sheath liquid is in a specified pH value range, ensures that cell screening operation is smoothly carried out, and can automatically complete the measurement of the pH value of the sheath liquid in the operation process of the stirring assembly through the design of the pH measuring part.

In summary, the invention can automatically complete stirring, bubble removal and pH value measurement treatment of the sheath liquid by utilizing the change of the sheath liquid flow in the sheath liquid flow process, ensures uniform mixing of the sheath liquid components, avoids influence on cell screening results caused by mixing of bubbles in the sheath liquid into a flow chamber, ensures cell screening effect, and has high overall automation degree and simple structure.

Drawings

FIG. 1 is a schematic diagram of a cell screening apparatus according to the present invention;

FIG. 2 is an enlarged schematic view of the sheath fluid inlet tube and its connection assembly shown in FIG. 1;

FIG. 3 is a schematic view of the structure of FIG. 2 after being deflected downward by a certain angle;

FIG. 4 is a schematic view of the structure of FIG. 3 with the flow chamber and its connection components removed;

FIG. 5 is a schematic view of the internal structure of the buffer tube of FIG. 4;

FIG. 6 is an enlarged schematic view of the structure of the buffer tube and its upper end assembly of FIG. 5;

FIG. 7 is an enlarged schematic view of the structure of FIG. 6 with the buffer tube removed;

FIG. 8 is an enlarged schematic view of the venturi and its components of FIG. 7;

FIG. 9 is a schematic view of the back structure of FIG. 8;

fig. 10 is an enlarged schematic view of the stationary ring gear and its connection assembly of fig. 8.

In the figure: 1. a body; 2. a sheath fluid inlet pipe; 3. a sheath fluid outlet pipe; 4. a cell suspension tube; 5. a nozzle ultrasonic oscillator; 6. a flow chamber; 7. a buffer tube; 8. a partition plate; 9. a conduit; 10. a tubule; 11. a venturi tube; 12. a rotating rod; 13. an impeller; 14. a one-way exhaust pipe; 15. a rotating shaft; 16. exhaust fan blades; 17. a belt drive structure; 18. a through hole; 19. a pH probe; 20. a fixed block; 21. an electric push rod; 22. a seal ring; 23. a support rod; 24. a chute; 25. a T-shaped block; 26. a fixed rack; 27. a touch block; 28. a bump; 29. a touch switch; 30. fixing the gear ring; 31. and (3) a spring.

Detailed Description

Referring to fig. 1, a cell screening apparatus includes a sheath fluid inlet tube 2, a cell suspension tube 4, and a flow chamber 6 mounted on a body 1, and further includes:

The buffer part is used for stirring the sheath liquid by using the input pressure of the sheath liquid and providing driving for bubble removal operation, the buffer part comprises a buffer tube 7 fixedly communicated with the sheath liquid inlet tube 2, the bubble removal part is used for removing bubbles existing in the sheath liquid, the bubble removal part comprises a guide tube 9 fixedly communicated with the buffer tube 7 and a thin tube 10, the pH measuring part is used for measuring the pH value of the sheath liquid, and the pH measuring part comprises a pH probe 19.

The one end that sheath liquid feed tube 2 was kept away from to buffer tube 7 is fixed to be linked together there is sheath liquid exit tube 3, the fixed intercommunication of sheath liquid exit tube 3 other end has nozzle ultrasonic oscillator 5, and nozzle ultrasonic oscillator 5 and flow chamber 6 fixed intercommunication, the sheath liquid gets into buffer tube 7 through sheath liquid feed tube 2, get into nozzle ultrasonic oscillator 5 through sheath liquid exit tube 3 after stirring and the bubble removal is handled, thoroughly mix the various components in the sheath liquid and evenly suspend under nozzle ultrasonic oscillator 5's effect, ensure cell suspension and sheath liquid intensive mixing in the experimentation, nozzle ultrasonic oscillator 5 can also carry out further bubble processing work to the sheath liquid simultaneously, guarantee the degree of accuracy of testing result.

Referring to fig. 1 to 10, a partition plate 8 is fixedly installed in the buffer tube 7, the partition plate 8 divides the buffer tube 7 into two parts, a part close to the sheath liquid inlet tube 2 is an inlet cavity, a part close to the sheath liquid outlet tube 3 is an outlet cavity, sheath liquid entering the buffer tube 7 through the sheath liquid inlet tube 2 is accumulated in the inlet cavity, then enters the outlet cavity through a guide tube 9, a venturi tube 11 and a thin tube 10, then enters the nozzle ultrasonic oscillator 5 through the sheath liquid outlet tube 3 and is mixed with cell suspension, the sheath liquid inlet tube 2 and the guide tube 9 are both positioned on the left side of the partition plate 8, the thin tube 10 and the sheath liquid outlet tube 3 are both positioned on the right side of the partition plate 8, and the diameter of the guide tube 9 is larger than that of the thin tube 10.

The fixed intercommunication has venturi 11 between pipe 9 and the tubule 10, venturi 11's design can make the sheath liquid flow rate increase that flows into tubule 10 from pipe 9, and install stirring subassembly on the venturi 11, stirring subassembly is including sealed rotation connection bull stick 12 on venturi 11, the one end fixed mounting that bull stick 12 is located venturi 11 has a plurality of impellers 13, when sheath liquid flows at great velocity of flow, accessible is to the impact of impeller 13 make bull stick 12 rotate, the sheath liquid can carry out certain stirring effect to sheath liquid with the impact of impeller 13 simultaneously, and its velocity of flow also changes when sheath liquid and impeller 13 impact, and the velocity of flow change of this part also can make the meeting part form a mixing zone, the sheath liquid in the mixing zone also has fluctuation, improve the interior composition mixing effect of sheath liquid.

The unidirectional exhaust pipe 14 is fixedly communicated on the tubule 10, gas allowed by the action of the unidirectional exhaust pipe 14 can be exhausted outwards only through the unidirectional exhaust pipe 14, an exhaust component is installed on the unidirectional exhaust pipe 14 and comprises a rotating shaft 15 which is connected to the unidirectional exhaust pipe 14 in a sealing and rotating mode, a plurality of exhaust fan blades 16 are fixedly installed at one end of the rotating shaft 15, which is located in the unidirectional exhaust pipe 14, a belt transmission structure 17 is installed between the venturi 11 and one end, which is located outside the unidirectional exhaust pipe 14, of the rotating shaft 12, when the rotating shaft 12 rotates due to the impeller 13, the rotating shaft 15 rotates simultaneously through the action of the belt transmission structure 17, negative pressure is generated in the unidirectional exhaust pipe 14 at the moment, bubbles in sheath liquid are exhausted outwards through the unidirectional exhaust pipe 14, and a bubble removal effect is achieved.

The belt transmission structure 17 is composed of a belt pulley I, a belt pulley II and a belt, the belt pulley I is fixedly arranged on the rotary rod 12, the belt pulley II is fixedly arranged on the rotary shaft 15, the belt is sleeved between the belt pulley I and the belt pulley II, the diameter of the belt pulley I is larger than that of the belt pulley II, and the speed is higher when the rotary rod 12 drives the rotary shaft 15 to rotate through the diameter difference between the belt pulley I and the belt pulley II, so that the exhaust effect of the unidirectional exhaust pipe 14 can be improved to a certain extent.

The electric push rod 21 is fixedly arranged on the buffer tube 7, the fixing block 20 is fixedly arranged between the output end of the electric push rod 21 and the pH probe 19, the pH probe 19 can be inserted into the buffer tube 7 during operation through the design of the electric push rod 21, the pH of sheath liquid in the buffer tube 7 is measured by using the electric push rod, and the pH probe 19 is positioned on the right side of the partition plate 8, so that the measured pH value is more accurate after the sheath liquid is fully mixed and de-bubbling.

The buffer tube 7 is provided with the through hole 18, the through hole 18 is internally and fixedly provided with the sealing ring 22 which is in sealing sliding connection with the pH probe 19, and under the design of the sealing ring 22, the sealing effect between the through hole 18 and the pH probe 19 can be ensured, and the outside air is prevented from moving into the buffer tube 7 due to the pH probe 19.

The buffer tube 7 is provided with a control component matched with the electric push rod 21, the control component is matched with the stirring component, the control component comprises a support rod 23 fixedly arranged on the buffer tube 7, a fixed rack 26 is slidably arranged on the support rod 23, one end of the rotating rod 12, which is far away from the belt wheel I, is fixedly provided with a fixed gear ring 30, the fixed gear ring 30 is meshed with the fixed rack 26, the support rod 23 is fixedly provided with a bump 28, the bottom of the bump 28 is fixedly provided with a touch switch 29 electrically connected with the electric push rod 21, the top of the fixed rack 26 is fixedly provided with a touch block 27 corresponding to the position of the touch switch 29, when the touch switch 29 is touched by the touch block 27, the electric push rod 21 starts to shrink so that the pH probe 19 is inserted into the buffer tube 7, otherwise, when the touch switch 29 is not touched, the electric push rod 21 keeps an extension state so that the pH probe 19 is positioned outside the buffer tube 7.

The trigger switch 29 and the pH probe 19 are all of the prior art, and the working principle and specific structure thereof are not described herein.

The T-shaped block 25 is fixedly arranged on the side wall of the fixed rack 26, the chute 24 matched with the T-shaped block 25 is arranged on the side wall of the supporting rod 23, the spring 31 is arranged between the chute 24 and the T-shaped block 25, the connection stability between the fixed rack 26 and the supporting rod 23 can be ensured under the matching of the chute 24 and the T-shaped block 25, and the touch switch 29 can be kept in an untouched state under the action of the spring 31 when the fixed rack 26 is not subjected to external force.

The maximum distance between the touch switch 29 and the touch block 27 is equal to the length of the fixed rack 26, and the size has the design advantage that when the fixed rack 26 is pushed by the fixed gear ring 30 to move upwards, the touch effect on the touch switch 29 can be ensured at the maximum moving position, and meanwhile, the fixed rack 26 and the fixed gear ring 30 are in a state to be separated, so that the fixed gear ring 30 is not subjected to larger rotation resistance due to limited movement of the fixed rack 26 when continuously rotating, and continuous and smooth rotation of the rotating rod 12 is ensured.

In the invention, the working principle of a cell screening device comprises the following steps:

Sample preparation: collecting cell samples, carrying out necessary pretreatment such as cell culture, centrifugation and the like to ensure the quality and purity of the samples, suspending the cell samples in a proper buffer solution to support the cell suspension, and taking a certain volume of sheath liquid for standby;

Screening: cell suspension is input into a flow chamber 6 through a cell suspension pipe 4, sheath fluid is input into a buffer pipe 7 through a sheath fluid inlet pipe 2, and after buffer treatment of the buffer pipe 7, the cell suspension enters a nozzle ultrasonic oscillator 5 through a sheath fluid outlet pipe 3 to be mixed with the cell suspension;

Under the action of the sheath fluid, the cells sequentially pass through a laser system in the machine body 1 one by one, the laser beams irradiate the cells one by one, the markers of the cells are excited to emit fluorescent signals, the optical system captures and records the signals, and the cells are identified and classified according to the intensity and fluorescent color of the signals;

The cells are classified and collected under the action of different charge deflection plates through a laser system, so that screening treatment is realized;

Bubble removal treatment of sheath liquid: sheath liquid entering the buffer tube 7 enters the thin tube 10 from the venturi tube 11 through the guide tube 9, when the sheath liquid flows through the venturi tube 11, the flow speed of the sheath liquid is increased, at the moment, the impeller 13 is impacted by the sheath liquid to enable the rotating rod 12 to rotate, meanwhile, the rotation of the impeller 13 and the impact effect of the sheath liquid can fully mix the sheath liquid, so that the internal components of the sheath liquid are uniformly distributed, and meanwhile, the precipitation of bubbles in the sheath liquid is facilitated;

When the rotating rod 12 rotates, the rotating shaft 15 is driven to rotate at a high speed through the belt transmission structure 17, and at the moment, bubbles separated out from sheath liquid under the action of the plurality of exhaust fan blades 16 are discharged through the unidirectional exhaust pipe 14, so that bubble removal operation is realized;

And (3) measuring the pH value of the sheath liquid: during the rotation of the rotating rod 12, the fixed gear ring 30 rotates simultaneously, at this time, the fixed gear rack 26 meshed with the fixed gear ring 30 moves upwards along the supporting rod 23 and drives the upper touch block 27 to touch the touch switch 29, at this time, the touch switch 29 sends out an electric signal to enable the electric push rod 21 to shrink, the pH probe 19 is inserted into the buffer tube 7, and the measurement treatment of pH is carried out on the sheath fluid after the complete mixing and bubble removal.

The foregoing has shown and described the basic principles and main features of the present invention and the advantages of the present invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined in the appended claims.

Claims

1. Cell sieving mechanism, including installing sheath liquid feed tube (2), cell suspension pipe (4) and flow chamber (6) on organism (1), its characterized in that still includes:

The buffer part comprises a buffer tube (7) fixedly communicated with the sheath liquid inlet tube (2), one end, far away from the sheath liquid inlet tube (2), of the buffer tube (7) is fixedly communicated with a sheath liquid outlet tube (3), the other end of the sheath liquid outlet tube (3) is fixedly communicated with a nozzle ultrasonic oscillator (5), the nozzle ultrasonic oscillator (5) is fixedly communicated with the flow chamber (6), and a baffle plate (8) is fixedly arranged in the buffer tube (7);

The device comprises a bubble removing part, a buffer tube (7) and a gas inlet pipe, wherein the bubble removing part comprises a guide tube (9) and a thin tube (10) which are fixedly communicated with the buffer tube (7), a venturi tube (11) is fixedly communicated between the guide tube (9) and the thin tube (10), an agitating assembly is arranged on the venturi tube (11), a unidirectional exhaust pipe (14) is fixedly communicated on the thin tube (10), and an exhaust assembly is arranged on the unidirectional exhaust pipe (14);

The pH measuring part comprises a pH probe (19), an electric push rod (21) is fixedly installed on the buffer tube (7), a fixed block (20) is fixedly installed between the output end of the electric push rod (21) and the pH probe (19), a control component matched with the electric push rod (21) is installed on the buffer tube (7), and the control component is matched with the stirring component.

2. The cell screening device according to claim 1, wherein the sheath fluid inlet tube (2) and the catheter (9) are both positioned on the left side of the partition plate (8), the tubule (10) and the sheath fluid outlet tube (3) are both positioned on the right side of the partition plate (8), and the diameter of the catheter (9) is larger than that of the tubule (10).

3. A cell screening apparatus according to claim 1, wherein the agitation assembly comprises a rotating rod (12) sealingly and rotatably connected to the venturi tube (11), and wherein a plurality of impellers (13) are fixedly mounted to one end of the rotating rod (12) located in the venturi tube (11).

4. A cell screening apparatus according to claim 3, wherein the exhaust assembly comprises a rotary shaft (15) sealingly and rotatably connected to the unidirectional exhaust pipe (14), and a plurality of exhaust fan blades (16) are fixedly mounted at one end of the rotary shaft (15) located in the unidirectional exhaust pipe (14).

5. A cell screening device according to claim 4, wherein a belt transmission structure (17) is mounted between the rotating rod (12) and the end of the rotating shaft (15) outside the venturi tube (11) and the unidirectional exhaust tube (14).

6. The cell screening apparatus according to claim 5, wherein the belt transmission structure (17) is composed of a pulley one fixedly mounted on the rotating rod (12), a pulley two fixedly mounted on the rotating shaft (15), and a belt sleeved between the pulley one and the pulley two, and the pulley one has a diameter larger than that of the pulley two.

7. The cell screening device according to claim 6, wherein the control assembly comprises a supporting rod (23) fixedly mounted on the buffer tube (7), a fixed rack (26) is slidably mounted on the supporting rod (23), a fixed gear ring (30) is fixedly mounted at one end, far away from the first pulley, of the rotating rod (12), the fixed gear ring (30) is meshed with the fixed rack (26), a protruding block (28) is fixedly mounted on the supporting rod (23), a touch switch (29) electrically connected with the electric push rod (21) is fixedly mounted at the bottom of the protruding block (28), and a touch block (27) corresponding to the position of the touch switch (29) is fixedly mounted at the top of the fixed rack (26).

8. The cell screening device according to claim 7, wherein a T-shaped block (25) is fixedly mounted on the side wall of the fixed rack (26), a chute (24) matched with the T-shaped block (25) is formed on the side wall of the supporting rod (23), and a spring (31) is mounted between the chute (24) and the T-shaped block (25).

9. A cell screening device according to claim 7, wherein the maximum distance between the trigger switch (29) and the trigger block (27) is equal to the length of the stationary rack (26).

10. Cell screening device according to claim 1, characterized in that the buffer tube (7) is provided with a through hole (18), and a sealing ring (22) in sealing sliding connection with the pH probe (19) is fixedly arranged in the through hole (18).