Single cell operation device

Abstract

The application discloses a single cell operation device, comprising: an objective table for fixing a container for bearing a sample; the probe assembly comprises a probe and a clamping device used for clamping the probe, and the probe is provided with a front end and a rear end; the control assembly comprises a fixed-point rotating device and a moving platform for moving the probe assembly in space to adjust the position of the probe, the fixed-point rotating device comprises an arc-shaped guide rail, a sliding seat, a locking assembly and a gap eliminating assembly, one of the arc-shaped guide rail and the sliding seat is provided with a connecting seat for fixing the probe assembly, and the other one is linked to the moving platform; in the relative movement process of the sliding seat and the arc-shaped guide rail, the front end of the probe is always located at the same position in a three-dimensional space; a microscopic imaging assembly; and the air pressure source assembly provides positive or negative air pressure for the probe. The probe assembly is restrained through the fixed-point rotating device, and the relative position of the front end of the probe can be ensured in the process of adjusting the posture of the probe, so that cells are better extracted and released, and the operation experience is improved.

Description

[0001] This application is a divisional application of the patent application with the application number 202510073452.0;

[0002] The information of the patent application with the application number 202510073452.0 is as follows:

[0003] Title of the Invention: Single-cell Manipulation Device;

[0004] Application Date: January 17, 2025. Technical Field

[0005] This application relates to the technical field of cell manipulation, and particularly to a single-cell manipulation device. Background Art

[0006] When conducting cell research, it is necessary to observe the specific morphology of cells and extract the target cells separately for research. The currently common method for single-cell manipulation is in-situ extraction, specifically by sucking a single cell into a holding needle for spatial position transfer. During the operation, the front end opening of the microtube probe used for cell extraction is about 1 to 20 micrometers, and the extraction and release of a single cell are achieved through the air pressure change inside the microtube.

[0007] During the process of extracting and releasing cells, relative movement between the probe and the target position needs to be achieved. In the prior art, generally, the operating table carrying the cell container and the probe move relative to each other in three dimensions to achieve precise control at the microscopic scale. However, in addition to the movement in three dimensions, in actual operation, it is often necessary to adjust the posture of the probe to better extract and release cells. This causes trouble for the microscopic control of the front end of the probe, and it is possible that the front end of the probe after adjusting the posture deviates from the microscopic field of view, seriously affecting the operation efficiency and posing a hidden danger of damaging the probe or cells. Summary of the Invention

[0008] To solve the above technical problems, this application discloses a single-cell manipulation device, including:

[0009] A stage for fixing and carrying a container for the sample;

[0010] A probe assembly including a probe and a holder for clamping the probe. The probe is an elongated hollow structure with opposite front and rear ends. During use, the front end of the probe extends into the container, and the rear end of the probe is inserted into the holder;

[0011] The control component includes a fixed-point rotation device and a moving platform for moving the probe assembly in space to adjust the position of the probe. The fixed-point rotation device includes a mutually cooperating arc-shaped guide rail, a sliding seat, and a locking component for locking the relative positions of the two. A connecting seat for fixing the probe assembly is provided on one of the arc-shaped guide rail and the sliding seat, and the other is linked to the moving platform; during the relative movement of the sliding seat and the arc-shaped guide rail, the front end of the probe is always in the same position in three-dimensional space;

[0012] The microscopic imaging component provides a window for positioning the probe;

[0013] The air pressure source component is connected to the rear end of the probe via the gripper to provide positive or negative air pressure for the probe.

[0014] The following also provides several optional ways, which are not additional limitations to the above overall solution, but only further supplements or optimizations. Without technical or logical contradictions, each optional way can be combined with the above overall solution alone, or multiple optional ways can be combined with each other.

[0015] In one embodiment, the extension path of the arc-shaped guide rail is located in a plane, and the front end of the probe is located at the center position of the circle; or

[0016] The extension path of the arc-shaped guide rail is located on the conical surface of a cone, and the front end of the probe is located at the vertex position of the cone.

[0017] In one embodiment, the arc-shaped guide rail cooperates with the sliding seat through two opposite side edges, and the two opposite side edges are respectively located on both sides of the arc-shaped guide rail in the extending direction of the probe.

[0018] In one embodiment, a backlash elimination component is further provided between the arc-shaped guide rail and the sliding seat. The backlash elimination component includes a sliding piece provided on the side edge and an adjusting member provided on the arc-shaped guide rail or the sliding seat. The adjusting member can adjust its own position relative to the side edge and press against the sliding piece.

[0019] In one embodiment, the sliding piece extends in the same direction as the side edge, and a plurality of adjusting members are provided at intervals in the extending direction of the sliding piece.

[0020] In one embodiment, the arc-shaped guide rail has a front surface facing the probe and an opposite back surface. The sliding seat has a C-shaped cross-section and surrounds the arc-shaped guide rail. The front surface is received inside the C-shape of the sliding seat, and both ends of the C-shaped opening bypass the side edges of the arc-shaped guide rail and are close to the back surface.

[0021] In one embodiment, the sliding seat is provided with a cavity facing the front side and a plurality of sliding blocks arranged at intervals around the cavity, and the sliding seat is matched with the front side through the sliding blocks;

[0022] A position mark for marking the position of the probe is provided on the front side.

[0023] In one embodiment, the connecting seat includes a clamping member located radially of the gripper and a locking member cooperating with the clamping member. The clamping member forms a clamping area, the gripper is located in the clamping area, and the locking member extends into the clamping area and positions the gripper.

[0024] In one embodiment, an adjustable positioning ring is provided on the gripper, and the positioning ring abuts against the clamping member to pre-position the relative position of the gripper relative to the connecting seat.

[0025] In one embodiment, the gripper includes a housing, the housing includes a head and a body that are axially relatively movable, and the rear end of the probe passes through the head and extends into the body; a limiting portion is provided inside the body, the probe extends into the body and the rear end is positioned at the limiting portion, and a pressure channel communicating with the inside of the probe is provided on the limiting portion.

[0026] The technical solution disclosed in the present application realizes the constraint of the probe assembly through a fixed-point rotation device, and can ensure the relative position of the front end of the probe during the process of adjusting the posture of the probe, so as to realize better extraction and release of cells and improve the operation experience.

[0027] Specific beneficial technical effects will be further explained in combination with specific structures or steps in the specific implementation manner. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is a schematic diagram of a single-cell operation device according to an embodiment of the present application;

[0029] Figure 2 is a schematic diagram of the structure of the fixed-point rotation device and the probe assembly of the single-cell operation device;

[0030] Figure 3 is Figure 2 a schematic diagram of the structure of the fixed-point rotation device and the probe assembly in another perspective in;

[0031] Figure 4 is a schematic diagram of the structure of the probe assembly in different postures;

[0032] Figure 5 is a schematic diagram of the cooperation between the probe assembly and the fixed-point rotation device;

[0033] Figure 6 isFigure 2 Structural schematic diagram of the central fixed-point rotation device and the probe assembly from a top-down perspective;

[0034] Figure 7 is Figure 6 Partial schematic diagram from the cross-sectional perspective at AA in the figure;

[0035] Figure 8 Internal mating schematic diagram of the probe assembly;

[0036] Figure 9 Schematic diagram of the air pressure source assembly of the single-cell operation device;

[0037] Figure 10 Working schematic diagram of the air pressure source assembly.

[0038] Explanation of the reference numerals in the figure is as follows:

[0039] 10. Stage; 21. Optical microscope objective; 22. Image detection unit; 23. Light source; 50. Moving platform; 60. Air pressure source assembly; 61. Plunger pump; 61a. Inlet end; 61b. Outlet end; 61c. Plunger; 62. Solenoid valve; 62a. Common end; 62b. Normally open end; 62c. Normally closed end; 63. Three-way joint; 64. First one-way valve; 65. Second one-way valve; 66. Air filter; 67. Silencer; 68. Plug; 70. Control system;

[0040] 100. Probe;

[0041] 200. Holder; 211. Head; 212. Body; 2122. Limiting part; 2123. Pressure channel; 2124. First cylinder; 2125. Second cylinder; 2126. Third cylinder; 2127. Clamping end; 2128. Pressure end; 213. Positioning ring;

[0042] 300. Fixed-point rotation device; 301. Base; 310. Arc-shaped guide rail; 311. Limiting end; 312. Positioning pin; 313. Positioning hole; 320. Sliding seat; 321. Positioning piece; 322. Cavity; 323. Sliding block; 331. Locking rod; 340. Backlash elimination assembly; 341. Sliding piece; 342. Adjusting piece; 350. Connecting seat; 351. Clamping piece; 352. Locking piece. Specific embodiments

[0043] Next, the technical solutions in the embodiments of the present application will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present application.

[0044] It should be noted that when a component is referred to as being "connected" to another component, it can be directly connected to the other component or there may be an intermediate component. When a component is considered to be "disposed on" another component, it can be directly disposed on the other component or there may be an intermediate component at the same time.

[0045] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the technical field to which this application belongs. The terms used in the specification of this application herein are only for the purpose of describing specific embodiments and are not intended to limit this application. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0046] Refer to the attached Figure 1 to the attached Figure 4 As shown, this application discloses a single-cell manipulation device, including a stage for fixing and carrying a container for the sample, a probe assembly for operating on the sample, a control assembly for controlling the relative movement of the probe assembly, a microscopic imaging assembly for providing a window for positioning the probe 100, and a pneumatic source assembly for providing positive or negative air pressure for the probe 100.

[0047] The probe assembly includes a probe 100 and a holder 200 for clamping the probe 100. The probe 100 is an elongated hollow structure, and the material is quartz, glass, etc. The probe 100 can be made by laser heating and drawing of a quartz capillary and is a geometrically rotationally symmetric structure, with a hollow interior and no movable parts or embedded circuits. The probe 100 has opposite front and rear ends, and both the front and rear ends are open structures. The size of the rear end is adapted to the holder 200, and the pneumatic source assembly communicates with the rear end of the probe 100 via the holder 200. During use, the front end of the probe 100 extends into the container, the rear end of the probe 100 is inserted into the holder 200, the aperture of the front end is relatively reduced compared to the rear end, and it has a flat micro-operation end face to ensure that the cells will not be damaged.

[0048] The control assembly includes a fixed-point rotation device 300 and a moving platform 50 for moving the probe assembly in space to adjust the position of the probe 100. The fixed-point rotation device 300 includes a mutually cooperating arc-shaped guide rail 310, a sliding seat 320, and a locking assembly for locking the relative positions of the two. A connecting seat 350 for fixing the probe assembly is provided on one of the arc-shaped guide rail 310 and the sliding seat 320, and the other is linked to the moving platform 50; during the relative movement of the sliding seat 320 and the arc-shaped guide rail 310, the front end of the probe 100 always remains at the same position in three-dimensional space;

[0049] The technical solution disclosed in this application can achieve the movement of the probe 100 relative to the sample through the control component. Among them, the moving platform 50 and the fixed-point rotating device 300 of the control component are responsible for the movement in different dimensions respectively. For example, the moving platform 50 realizes the movement of the probe component in three-dimensional space, and the fixed-point rotating device 300 can realize the attitude adjustment of the probe component relative to the moving platform 50. Further, the fixed-point rotating device 300 can ensure the relative position of the front end of the probe 100 during the process of adjusting the attitude of the probe 100, so as to achieve better extraction and release of cells and improve the operation experience.

[0050] The arc-shaped guide rail 310 and the sliding seat 320 can achieve the fixed-point adjustment of the front end of the probe 100 through various cooperation methods. For example, the extension path of the arc-shaped guide rail 310 is located on the conical surface of a cone, and the front end of the probe 100 is located at the vertex position of the cone. The cone can be an actual component or an imaginary virtual body. It can also be, for example, Figure 4 In the illustrated embodiment, the extension path of the arc-shaped guide rail 310 is located in a plane, and the front end of the probe 100 is located at the center of the arc-shaped guide rail 310. Further, the distance between the connecting seat 350 and the front end of the probe 100 is equal to the radius of the arc-shaped guide rail 310.

[0051] It can be understood that in different embodiments, the arc-shaped guide rail 310 and the sliding seat 320 can be interchanged. Therefore, the following will combine the setting method in which the connecting seat 350 is arranged on the sliding seat 320 and the arc-shaped guide rail 310 is connected to the moving platform 50 through the base 301 in the attached Figure 2 to the attached Figure 7 to elaborate on the cooperation details between the two. In other embodiments, the connecting seat 350 can also be arranged on the arc-shaped guide rail 310, and the sliding seat 320 is connected to the moving platform 50. Those skilled in the art can clearly understand the corresponding technical solutions without any doubt and will not be elaborated below.

[0052] Attached Figure 2 to attached Figure 7 In the figures, the arc-shaped guide rail 310 cooperates with the sliding seat 320 through two opposite side edges. The two opposite side edges are respectively located on both sides of the arc-shaped guide rail 310 in the extending direction of the probe 100 (such as the up and down direction in the figure). In order to improve the adjustment accuracy of the probe 100 while reducing the requirements for the machining accuracy of the arc-shaped guide rail 310 and the sliding seat 320, in the attached Figure 5 and attached Figure 7In the illustrated embodiment, a backlash eliminating component 340 is further provided between the arc-shaped guide rail 310 and the sliding seat 320. The backlash eliminating component 340 includes a sliding piece 341 disposed on the side edge and an adjusting member 342 disposed on the arc-shaped guide rail 310 or the sliding seat 320. The adjusting member 342 can adjust its own position relative to the side edge and press against the sliding piece 341. In the figure, the sliding piece 341 extends in the same direction as the side edge. In the extending direction of the sliding piece 341, a plurality of adjusting members 342 are provided and arranged at intervals. In the figure, the adjusting member 342 can be movably disposed on the sliding seat 320 for positioning, such as being screwed. The adjusting member 342 can protrude or retract from the surface of the sliding seat 320 to change the relative position between the surface of the sliding seat 320 and the sliding piece 341. Each adjusting member 342 is adjusted independently. The sliding piece 341 can protect the side edge of the arc-shaped guide rail 310 to avoid wear while achieving uniform force application, ensuring the adjustment accuracy.

[0053] Refer to the attached Figure 5 to the attached Figure 7 In the illustrated embodiment, the arc-shaped guide rail 310 has a front face facing the probe 100 and an opposite back face. The cross-section of the sliding seat 320 is C-shaped and surrounds the arc-shaped guide rail 310. The front face is received inside the C-shaped cross-section of the sliding seat 320, and both ends of the opening of the C-shaped cross-section wind around the side edge of the arc-shaped guide rail 310 and are close to the back face. Among them, both ends of the opening of the C-shaped cross-section of the sliding seat 320 can be set to be in contact with the back face of the arc-shaped guide rail 310 or to have a gap with the back face of the arc-shaped guide rail 310. When both ends of the opening of the C-shaped cross-section are in contact with the back face of the arc-shaped guide rail 310, it can be set to be in contact without stress to provide a smooth sliding experience or to be in contact with mutual extrusion to achieve mutual positioning. In the figure, positioning pieces 321 are movably disposed at both ends of the C-shaped opening of the sliding seat 320. The sliding seat 320 cooperates with the back face of the arc-shaped guide rail 310 through the positioning pieces 321. Preferably, the positioning pieces 321 are provided in pairs and respectively correspond to the upper side edge and the lower side edge of the sliding seat 320. The positioning pieces 321 are connected to the body of the sliding seat 320 through fasteners and adjust the fitting clearance between the sliding seat 320 and the arc-shaped guide rail 310, such as the mutual contact or clearance fit mentioned above.

[0054] Further, the sliding seat 320 is provided with a cavity 322 facing the front and a plurality of spaced sliding blocks 323 around the cavity 322. The sliding blocks 323 are provided with smooth sliding surfaces facing the front, and the sliding seat 320 is engaged with the front through the sliding surfaces of the sliding blocks 323. The sliding blocks 323 can improve the positioning accuracy of the two while maintaining the sliding fit between the sliding seat 320 and the front, avoiding problems such as a decrease in positioning accuracy or excessive mating resistance caused by an overly large contact area. Corresponding thereto, a partial area of the front of the arc-shaped guide rail 310 is recessed relative to its side edge to form a recessed area, which corresponds to the cavity 322, and the sliding blocks 323 are engaged with the side edge. Further, a position mark for marking the position of the probe 100 is provided on the front, and the outer contour of the sliding seat 320 can cooperate with the position mark to indicate the position or attitude of the probe 100. The position mark is located in the recessed area to avoid being worn due to the relative movement between the sliding seat 320 and the arc-shaped guide rail 310.

[0055] The locking assembly also has various setting forms. For example, as shown in the reference attachment Figure 5 shown, the locking assembly includes a sliding piece 341 and a locking rod 331 pressing against the sliding piece 341, wherein the locking assembly can share the same sliding piece 341 with the backlash elimination assembly 340. The locking rod 331 is engaged with the middle part of the sliding piece 341, and at least the adjusting member 342 is located on both sides of the locking rod 331. The locking rod 331 can move relative to the sliding seat 320 to squeeze or release the sliding piece 341, thereby realizing the locking or release of the sliding seat 320 and the arc-shaped guide rail 310. The locking assembly can also be realized by the positioning piece 321 mentioned above. For example, by adjusting the relative distance between the positioning piece 321 and the body of the sliding seat 320 to realize clamping or releasing the arc-shaped guide rail 310, thereby realizing the locking or release of the sliding seat 320 and the arc-shaped guide rail 310. The setting methods of the locking assembly mentioned above can be implemented independently or cooperatively.

[0056] In addition to motion guidance, the arc-shaped guide rail 310 can also provide a limit for the sliding seat 320. Both ends of one side edge of the arc-shaped guide rail 310 protrude to form limit end portions 311, which abut against the limit end portions 311 when the sliding seat 320 moves to the extreme position. In the direction of gravity, the limit end portions 311 and the backlash elimination assembly 340 can be arranged on the opposite sides of the arc-shaped guide rail 310. For example, the limit end portions 311 are arranged on the side edge located below in the direction of gravity, and the backlash elimination assembly 340 is arranged on the side edge located above in the direction of gravity; the two can also be interchanged. Or in other embodiments, in the direction of gravity, the limit end portions 311 and the backlash elimination assembly 340 can also be arranged on the same side of the arc-shaped guide rail 310.

[0057] The arc-shaped guide rail 310 and the base 301 are positioned through the insertion and cooperation of the positioning pins 312 and the positioning holes 313, and are connected to each other through fasteners. The positioning holes 313 are arranged in pairs, and each pair of positioning holes 313 are respectively located on the arc-shaped guide rail 310 and the base 301. There are multiple positioning pins 312, which are respectively fitted into a pair of positioning holes 313. Among a pair of positioning holes 313, one positioning hole 313 located on the base 301 or on the arc-shaped guide rail 310 is an oval hole to allow the positioning pin 312 to finely adjust the relative position.

[0058] Regarding the mutual cooperation relationship between the connecting seat 350 and the gripper 200, reference can be made to the attached Figure 5 In the illustrated embodiment, the connecting seat 350 includes clamping members 351 located on both radial sides of the gripper 200 and a locking member 352 cooperating with the clamping members 351. The clamping members 351 form a clamping area, and the gripper 200 is located within the clamping area. The locking member 352 extends into the clamping area to position the gripper 200. Figure 5 In this case, the clamping members 351 are of C-shaped or O-shaped structure, and the hollow interior forms a clamping area. The locking member 352 penetrates the side wall of the clamping member 351 and enters the hollow interior, that is, the clamping area. The locking member 352 and / or the clamping member 351 can cooperate with the outer peripheral surface of the gripper 200 to realize the positioning of the gripper 200. Further, an adjustable positioning ring 213 is provided on the gripper 200, and the positioning ring 213 abuts against the clamping member 351 to pre-position the relative position of the gripper 200 relative to the connecting seat 350. In the direction of gravity, the positioning ring 213 cooperates with the upper end surface of the clamping member 351.

[0059] In order to ensure that the front end of the probe 100 is in a definite spatial position relative to the gripper 200, the gripper 200 in this embodiment can realize the precise positioning of the probe 100. Refer to the attached Figure 8 In the illustrated embodiment, the gripper 200 includes a housing, and the housing includes an axially relatively movable head 211 and a body 212. The rear end of the probe 100 passes through the head 211 and extends into the body 212; a limiting portion 2122 is provided inside the body 212. The probe 100 extends into the body 212 and the rear end is positioned at the limiting portion 2122. A pressure channel 2123 communicating with the inside of the probe 100 is provided on the limiting portion 2122. The limiting portion 2122 can be arranged as an annular structure inside the body 212. Regarding the specific implementation manner of the limiting portion 2122, in reference to an embodiment, the body 212 is of a multi-layer structure and includes:

[0060] A first cylinder 2124, located on the outer layer and open at both ends. One end is a clamping end 2127 extending into the head 211, and the other end is a pressure end 2128 connected to the peripheral device;

[0061] The second cylinder body 2125 is located inside the first cylinder body 2124. The second cylinder body 2125 includes an open end near the clamping end 2127 and a closed end located inside the first cylinder body 2124. The closed end forms a limiting part 2122;

[0062] The third cylinder body 2126 is located inside the first cylinder body 2124 and both ends are open. One end abuts against the closed end of the second cylinder body 2125, and the other end is close to the pressure end 2128 of the first cylinder body 2124.

[0063] By installing the second cylinder body 2125 and the third cylinder body 2126 inside the first cylinder body 2124, it is possible to ensure the positioning accuracy of the limiting part 2122 relative to the probe assembly while reducing the processing difficulty, thereby ensuring the spatial position of the front end of the probe 100 relative to the probe assembly, and thus cooperating with the fixed-point rotation device 300 to achieve the fixed-point adjustment of the front end of the probe 100.

[0064] In another embodiment, the air pressure source assembly 60 serves as a power source for sucking and discharging substances (such as cells, reagents, etc.). Refer to the attached Figure 9 In the illustrated embodiment, the air pressure source assembly includes a plunger pump 61, a solenoid valve 62, a three-way joint 63, a first one-way valve 64, a second one-way valve 65, an air filter 66, a silencer 67, and a plug 68. The plunger pump 61 has an inlet end 61a, an outlet end 61b, and a plunger 61c; the solenoid valve 62 has a common end 62a, a normally open end 62b, and a normally closed end 62c; the air filter 66 has an air inlet end and an air outlet end; the first one-way valve 64 has an "IN" port and an "OUT" port; the second one-way valve 65 has an "IN" port and an "OUT" port; the three-way joint 63 has three connection ports.

[0065] The plug 68 is screwed into the inlet end 61a of the plunger pump, tightened and sealed. The air intake and exhaust of the plunger pump are both completed through the outlet end 61b. The outlet end 61b of the plunger pump is connected to the common end 62a of the solenoid valve 62 by a pipeline; the air inlet end of the air filter 66 is connected and sealed to the silencer 67 by a threaded connection, and the air outlet end of the air filter 66 is connected to the "IN" port of the first one-way valve 64 by a pipeline; the "OUT" port of the first one-way valve 64 is connected to one of the ports of the three-way joint 63 by a pipeline. Among the other two ports of the three-way joint 63, one port is connected to the normally open end 62b of the solenoid valve 62 by a pipeline, and the other port is connected to the "IN" port of the second one-way valve 65 by a pipeline; the normally closed end 62c of the solenoid valve 62 is connected to the pressure end 2128 of the probe assembly through a pipeline, and further communicates with the probe 100, so that the entire air pressure source assembly is communicated with the probe 100.

[0066] The intake end of the air filter 66 is connected to a muffler 67 to reduce the noise of the gas path system of the entire operating cell. The exhaust end of the air filter 66 is connected to the "IN" port of the first one-way valve 64, and the first one-way valve 64 prevents the gas in the entire gas path system from flowing back to the air filter 66; the second one-way valve 65 is used to exhaust the air in the plunger pump 61 to ensure that there is no unfiltered air in the plunger pump 61.

[0067] Before operation, the common terminal 62a and the normally open terminal 62b of the solenoid valve 62 are first connected, and the normally closed terminal 62c is closed. The plunger 61c first moves upward to the uppermost position. During this process, the residual gas in the plunger pump 61 is exhausted through the tee joint 63 and the second one-way valve 65; then the plunger 61c moves downward to the middle position of the plunger pump 61. During this process, the air enters the plunger pump 61 through the air filter 66, the first one-way valve 64 and the tee joint 63. The initial stop position of the plunger 61c is set in the middle of the plunger pump, which can provide negative air pressure when moving downward and positive air pressure when moving upward.

[0068] In some embodiments, as Figure 10 shown, the plunger pump 61 provides positive or negative air pressure for the probe 100 in the form of pulses, and the plunger 61c moves intermittently within a single working stroke. When the plunger 61c moves upward intermittently, the moving distance each time is d1, and multiple positive air pressure pulses can be generated. The air pressure value of the positive air pressure pulse is P1, and the pulse period is T1; when the plunger 61c moves downward intermittently, the moving distance each time is d2, and multiple negative air pressure pulses can be generated. The air pressure value of the negative air pressure pulse is P2, and the pulse period is T2.

[0069] The origin O in the figure represents that the plunger 61c is in the initial position, and at this time the air pressure value in the plunger is approximately equal to the atmospheric pressure. Generally, the pulse air pressure value is selected as -10Kpa - 10Kpa, and the period is less than 1s, preferably 0.5s - 1s. The air pressure value and period of the pulse are adjustable to meet different actual needs.

[0070] The working principle of the air pressure source assembly is introduced below by taking the extraction of cells as an example:

[0071] The solenoid valve 62 is energized, the normally open terminal 62b is closed, and the normally closed terminal 62c is opened to connect the plunger pump 61 with the probe 100. The plunger 61c in the plunger pump 61 is controlled to move downward. At this time, the plunger pump 61 provides negative air pressure pulses for the probe 100, and the probe 100 can inhale the cells in the culture dish. If a single pulse cannot inhale the cells into the probe 100, multiple pulses can be continuously emitted until inhalation. During the process of the probe 100 inhaling the cells, when the internal and external air pressures are balanced, the cells stop moving, so they can be retained in the probe 100.

[0072] After the cell inhalation is completed, if the plunger 61c has not moved to the extreme position, the movement can be continued to provide pulses to inhale more cells.

[0073] When the plunger 61c moves downward to the extreme position, the normally closed end 62c of the solenoid valve 62 is closed, and the normally open end 62b is opened. The plunger 61c moves upward for reset. During the reset process, the gas in the plunger pump 61 is discharged through the three-way joint 63 and the second one-way valve 65. After the reset of the plunger 61c is completed, the normally open end 62b is closed and the normally closed end 62c is opened, and cells can be inhaled continuously. The operation is repeated in cycles to inhale more cells at different sites into the probe 100 in batches.

[0074] After the cell inhalation is completed, the plunger 61c is first reset (it can also not be reset), and the plunger 61c in the plunger pump 61 is controlled to move upward. At this time, a positive air pressure pulse is provided to the probe 100, and the probe 100 spits out cells into the culture dish. Since the positive air pressure is also provided in the form of pulses, the cells can be spit out one by one until all are spit out. The working process is opposite to the cell inhalation process and will not be elaborated here.

[0075] Using positive or negative air pressure pulses, it is possible to achieve spitting out or inhaling at most one cell per operation. Providing a negative air pressure pulse may cause insufficient suction, resulting in the cell not being able to enter the probe. By continuously outputting multiple pulses, the cell can eventually enter the probe 100. Spitting out cells is similar to this.

[0076] Since there is a relatively large space inside the probe, it can temporarily store more cells. The probe 100 can continuously inhale multiple cells and then spit them out at one time, significantly improving the operation efficiency. In addition, using the pulsed probe 100, cells can be spit out one by one, so multiple cells are distributed at different positions, eliminating the need for redistribution operations.

[0077] It can be seen that the solenoid valve 62 serves as the pressure balance device of the entire system. After the normally open end 62b is closed and the normally closed end 62c is opened, the plunger pump 61 does not work, and the air path system connecting the probe 100 and the plunger pump 61 can maintain pressure balance. The extracted cells can be completely stored in the probe 100, and then the plunger 61c can move again to generate new pulses. After the normally open end 62b is opened and the normally closed end 62c is closed, since the pressure has been balanced at this time, the reset operation of the plunger pump 61 does not affect the cells stored in the probe 100.

[0078] The microscopic imaging assembly includes a light source 23, an optical microscope objective 21, an image detection unit 22 (such as a CCD camera), etc. The light source is generally arranged directly above the stage 10, and the optical microscope objective 21 and the image detection unit 22 are located below the stage. The control system 70 can be a terminal device such as a computer. The plunger pump 61 in the pneumatic source assembly 60, the solenoid valve 62, the motor in the moving platform 50, and the motor in the stage 10 are controlled by this control system. The control system also has an imaging display unit, that is, the probe 100 and the target cells can be displayed on the screen through the imaging system, and the operator can perform real-time operations.

[0079] Other details of the single-cell manipulation device can be implemented in combination with the prior art and will not be elaborated here.

[0080] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as within the scope described in this specification. When the technical features in different embodiments are embodied in the same drawing, it can be regarded that the drawing also discloses the combination examples of the various embodiments involved.

[0081] The above-described embodiments merely represent several implementation manners of the present application, and the description thereof is relatively specific and detailed, but it should not be construed as a limitation on the scope of the patent application. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present application, several modifications and improvements can still be made, and these all belong to the protection scope of the present application.

Claims

1. A single-cell manipulation device, characterized in that, Comprising: A stage for fixing and carrying a container holding a sample; A probe assembly including a probe and a holder for clamping the probe. The probe is an elongated hollow structure with a front end and a rear end opposite to each other. During use, the front end of the probe extends into the container, and the rear end of the probe is inserted into the holder; A control assembly including a fixed-point rotation device and a moving platform for moving the probe assembly in space to adjust the position of the probe. The fixed-point rotation device includes a mutually cooperating arc-shaped guide rail, a sliding seat, a locking assembly for locking their relative positions, and a backlash eliminating assembly acting between them. A connecting seat for fixing the probe assembly is provided on one of the arc-shaped guide rail and the sliding seat, and the other is linked to the moving platform; during the relative movement of the sliding seat and the arc-shaped guide rail, the front end of the probe always remains at the same position in three-dimensional space; A microscopic imaging assembly providing a window for positioning the probe; A pneumatic source assembly, which is connected to the rear end of the probe via the holder to provide positive or negative air pressure for the probe.

2. The single-cell manipulation device according to claim 1, wherein The extending path of the arc-shaped guide rail is located in a plane, and the front end of the probe is located at the center of the arc-shaped guide rail; or The extending path of the arc-shaped guide rail is located on the conical surface of a cone, and the front end of the probe is located at the vertex of the cone.

3. The single-cell manipulation device according to claim 1, wherein The arc-shaped guide rail cooperates with the sliding seat through two opposite side edges, and the two opposite side edges are respectively located on both sides of the arc-shaped guide rail in the extending direction of the probe.

4. The single cell manipulation device according to claim 3, wherein The backlash eliminating assembly includes a sliding piece provided on the side edge and an adjusting member provided on the arc-shaped guide rail or the sliding seat. The adjusting member can adjust its own position relative to the side edge and press against the sliding piece.

5. The single-cell manipulation device according to claim 4, wherein The sliding piece extends in the same direction as the side edge, and a plurality of adjusting members are provided at intervals in the extending direction of the sliding piece.

6. The single-cell manipulation device according to claim 1, wherein The arc-shaped guide rail has a front face facing the probe and a back face opposite thereto. The sliding seat has a C-shaped cross-section and surrounds the arc-shaped guide rail. The front face is received inside the C-shape of the sliding seat, and both ends of the C-shaped opening bypass the side edge of the arc-shaped guide rail and are close to the back face.

7. The single cell manipulation device according to claim 6, wherein The sliding seat is provided with a cavity facing the front face and a plurality of spaced sliding blocks surrounding the cavity. The sliding seat cooperates with the front face through the sliding blocks; A position mark for marking the position of the probe is provided on the front face.

8. The single-cell manipulation device according to claim 1, wherein, The connecting seat includes a clamping member located radially of the holder and a locking member cooperating with the clamping member. The clamping member forms a clamping area, the holder is located in the clamping area, and the locking member extends into the clamping area and positions the holder.

9. The single-cell manipulation device according to claim 8, wherein An adjustable positioning ring is provided on the holder, and the positioning ring abuts against the clamping member to pre-position the relative position of the holder relative to the connecting seat.

10. The single-cell manipulation device according to claim 1, wherein The gripper includes a housing, the housing includes a head and a body that are axially relatively movable, and the rear end of the probe passes through the head and extends into the body; a limiting portion is provided inside the body, the probe extends into the body and the rear end is positioned at the limiting portion, and a pressure channel communicating with the inside of the probe is provided on the limiting portion.

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