Single cell manipulation device

Abstract

The present application discloses a single-cell manipulation device, including: a stage for fixing a container carrying a sample; a probe assembly, including a probe and a clamp for clamping the probe, the probe having a front end and a rear end; a control assembly, including a fixed-point rotation device and a mobile platform for moving the probe assembly in space to adjust the position of the probe, the fixed-point rotation device including an arc-shaped guide rail, a sliding seat and a locking 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 is linked to the mobile 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 the three-dimensional space; a microscopic imaging assembly; and an air pressure source assembly, which provides positive or negative air pressure for the probe. 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 in the process of adjusting the probe posture, thereby achieving better extraction and release of cells and improving the operating experience.

Description

Technical Field

[0001] The present application relates to the field of cell manipulation technology, and in particular to a single cell manipulation device. Background Art

[0002] When conducting cell research, it is necessary to observe the specific morphology of the cells and extract the target cells separately for research. The commonly used method for single-cell operation is in situ extraction, which specifically uses a suction needle to suck in a single cell for spatial position transfer. During the operation, the front end opening of the microtube probe used for cell extraction is about 1 to 20 microns, and the extraction and release of a single cell is achieved through changes in air pressure inside the microtube.

[0003] In the process of extracting and releasing cells, it is necessary to achieve relative movement between the probe and the target position. In the prior art, precise control on the microscopic size is generally achieved by relative movement of the operating table and the probe carrying the cell container in three dimensions. However, in addition to the movement in three dimensions, it is often necessary to adjust the posture of the probe in actual operation to better extract and release cells. This causes trouble for the microscopic control of the front end of the probe. The front end of the probe may be out of the microscopic field of view after the posture is adjusted, which seriously affects the operating efficiency and has the risk of damaging the probe or cells. Summary of the invention

[0004] In order to solve the above technical problems, the present application discloses a single cell operation device, comprising:

[0005] A stage for fixing the container carrying the sample;

[0006] A probe assembly, comprising a probe and a holder for holding the probe, wherein the probe is an elongated hollow structure having a front end and a rear end opposite to each other, wherein the front end of the probe extends into the container and the rear end of the probe is inserted into the holder when in use;

[0007] A control assembly, comprising a fixed-point rotating device and a mobile platform for moving the probe assembly in space to adjust the position of the probe, wherein the fixed-point rotating device comprises a mutually cooperating arc-shaped guide rail, a sliding seat, and a locking assembly for locking the relative positions of the two, wherein 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 is linked to the mobile 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 the three-dimensional space;

[0008] A microscopic imaging component, providing a window for positioning the probe;

[0009] An air pressure source component is connected to the rear end of the probe via the clamp to provide positive or negative air pressure for the probe.

[0010] Several optional methods are also provided below, but they are not intended to be additional limitations on the above-mentioned overall solution, but are merely further supplements or preferences. Under the premise that there are no technical or logical contradictions, each optional method can be combined with the above-mentioned overall solution separately, and multiple optional methods can also be combined.

[0011] 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 of the circle; or

[0012] The extension path of the arc-shaped guide rail is located on the conical surface of a vertebral body, and the front end of the probe is located at the apex position of the vertebral body.

[0013] 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 two sides of the arc-shaped guide rail in the extension direction of the probe.

[0014] In one of the embodiments, a gap-eliminating assembly is further provided between the arc guide rail and the sliding seat, and the gap-eliminating assembly includes a sliding plate arranged on the side edge and an adjusting member arranged on the arc guide rail or the sliding seat, and the adjusting member can adjust its own position relative to the side edge and press against the sliding plate.

[0015] In one embodiment, the sliding sheet and the side edge extend in the same direction, and a plurality of the adjusting members are provided and spaced apart in the extending direction of the sliding sheet.

[0016] In one embodiment, the arc guide rail has a front side facing the probe and an opposite back side, the sliding seat has a C-shaped cross-section and surrounds the arc guide rail, the C-shaped interior of the sliding seat accommodates the front side and both ends of the C-shaped opening pass through the side edges of the arc guide rail and are close to the back side.

[0017] In one embodiment, the sliding seat is provided with a cavity toward the front side and a plurality of sliding blocks are provided around the cavity at intervals, and the sliding seat cooperates with the front side through the sliding blocks;

[0018] A position mark for marking the position of the probe is provided on the front side.

[0019] In one embodiment, the connecting seat includes a clamping member located radially of the clamp and a locking member cooperating with the clamping member, the clamping member forms a clamping area, the clamp is located in the clamping area, and the locking member extends into the clamping area and positions the clamp.

[0020] In one embodiment, the clamp is provided with an adjustable positioning ring, and the positioning ring abuts against the clamping member to pre-position the relative position of the clamp with respect to the connecting seat.

[0021] In one embodiment, the clamp includes a shell, the shell includes a head and a body that are axially movable relative to each other, 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 the limiting portion is provided with a pressure channel connected to the interior of the probe.

[0022] The technical solution disclosed in the present application realizes the constraint of the probe assembly through a fixed-point rotating device, and can ensure the relative position of the front end of the probe during the process of adjusting the posture of the probe, thereby achieving better extraction and release of cells and improving the operating experience.

[0023] The specific beneficial technical effects will be further explained in the specific implementation methods in combination with specific structures or steps. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a schematic diagram of a single cell manipulation device in one embodiment of the present application;

[0025] Figure 2 This is a schematic diagram of the structure of the fixed-point rotating device and probe assembly of the single-cell manipulation device;

[0026] Figure 3 for Figure 2 A schematic structural diagram of the fixed-point rotating device and the probe assembly from another perspective;

[0027] Figure 4 It is a schematic diagram of the structure of the probe assembly in different postures;

[0028] Figure 5 It is a schematic diagram of the cooperation between the probe assembly and the fixed-point rotating device;

[0029] Figure 6 for Figure 2 A schematic diagram of the structure of the central fixed-point rotating device and the probe assembly from a top-down perspective;

[0030] Figure 7 for Figure 6 A partial schematic diagram from the cross-section perspective at AA in the middle;

[0031] Figure 8 It is a schematic diagram of the internal matching of the probe assembly;

[0032] Fig. 9 A schematic diagram of the air pressure source assembly of the single-cell manipulation device;

[0033] Fig.10This is a working schematic diagram of the air pressure source component.

[0034] The reference numerals in the figures are described as follows:

[0035] 10. stage; 21. optical microscope objective lens; 22. image detection unit; 23. light source; 50. mobile 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 connector; 64. first one-way valve; 65. second one-way valve; 66. air filter; 67. silencer; 68. plug; 70. control system;

[0036] 100. Probe;

[0037] 200, clamp; 211, head; 212, body; 2122, limiter; 2123, pressure channel; 2124, first cylinder; 2125, second cylinder; 2126, third cylinder; 2127, clamping end; 2128, pressure end; 213, positioning ring;

[0038] 300, fixed-point rotating device; 301, base; 310, arc-shaped guide rail; 311, limit end; 312, positioning pin; 313, positioning hole; 320, sliding seat; 321, positioning plate; 322, cavity; 323, sliding block; 331, locking rod; 340, anti-backlash assembly; 341, sliding plate; 342, adjusting member; 350, connecting seat; 351, clamping member; 352, locking member. DETAILED DESCRIPTION

[0039] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0040] It should be noted that when a component is referred to as being "connected" to another component, it may be directly connected to the other component or there may be a central component. When a component is referred to as being "disposed on" another component, it may be directly disposed on the other component or there may be a central component at the same time.

[0041] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application 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.

[0042] Reference Figure 1 To Attachment Figure 4 As shown, the present application discloses a single-cell manipulation device, including a stage for fixing a container carrying a sample, a probe assembly for manipulating the sample, a control assembly for controlling the relative movement of the probe assembly, a microscopic imaging assembly providing a window for positioning the probe 100, and an air pressure source assembly for providing positive or negative air pressure to the probe 100.

[0043] The probe assembly includes a probe 100 and a holder 200 for clamping the probe 100. The probe 100 is an elongated hollow structure made of quartz, glass, etc. The probe 100 can be formed by laser heating and drawing of a quartz capillary and has a geometric rotationally symmetrical structure. The interior is hollow and has no movable parts or embedded circuits. The probe 100 has a relative front end and a rear end, and both the front end and the rear end are open structures. The size of the rear end is adapted to the holder 200, and the air pressure source assembly is connected to the rear end of the probe 100 via the holder 200. When in use, the front end of the probe 100 extends into the container, and the rear end of the probe 100 is inserted into the holder 200. The aperture of the front end is reduced relative to the rear end, and has a flat micro-operation end face to ensure that the cells will not be damaged.

[0044] The control assembly includes a fixed-point rotating device 300 and a mobile platform 50 for moving the probe assembly in space to adjust the position of the probe 100. The fixed-point rotating device 300 includes an arc-shaped guide rail 310, a sliding seat 320 and a locking assembly for locking the relative positions of the two. One of the arc-shaped guide rail 310 and the sliding seat 320 is provided with a connecting seat 350 for fixing the probe assembly, and the other is linked to the mobile 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 is always in the same position in the three-dimensional space.

[0045] The technical solution disclosed in the present application can realize the movement of the probe 100 relative to the sample through the control component. The mobile 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 mobile platform 50 realizes the movement of the probe component in three-dimensional space, and the fixed-point rotating device 300 can realize the posture adjustment of the probe component relative to the mobile platform 50. Furthermore, 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 posture of the probe 100, thereby achieving better extraction and release of cells and improving the operating experience.

[0046] The arc guide rail 310 and the sliding seat 320 can realize the fixed-point adjustment of the front end of the probe 100 through various matching modes. For example, the extension path of the arc guide rail 310 is located on the conical surface of a vertebra, and the front end of the probe 100 is located at the apex of the vertebra. The vertebra can be an actual component or an imaginary virtual body. Figure 4 In the illustrated embodiment, the extension path of the arc guide rail 310 is located in a plane, the front end of the probe 100 is located at the center of the arc guide rail 310 , and the distance between the connecting seat 350 and the front end of the probe 100 is equal to the radius of the arc guide rail 310 .

[0047] It is understandable that in different embodiments, the arc guide rail 310 and the sliding seat 320 can be interchanged. Figure 2 To Attachment Figure 7 The middle connecting seat 350 is arranged on the sliding seat 320, and the arc guide rail 310 is connected to the mobile platform 50 through the base 301 to illustrate the coordination details of the two. In other embodiments, the connecting seat 350 can also be arranged on the arc guide rail 310, and the sliding seat 320 is connected to the mobile platform 50. Those skilled in the art can clearly identify the corresponding technical solutions without any doubt, which will not be repeated below.

[0048] Attached Figure 2 To Attachment Figure 7 In the embodiment, the arc guide rail 310 cooperates with the sliding seat 320 through two opposite side edges, and the two opposite side edges are respectively located on both sides of the arc guide rail 310 in the extension direction of the probe 100 (for example, 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 processing accuracy of the arc guide rail 310 and the sliding seat 320, the present application is attached. Figure 5 and attached Figure 7 In the illustrated embodiment, a clearance elimination assembly 340 is further provided between the arc guide rail 310 and the sliding seat 320. The clearance elimination assembly 340 includes a sliding piece 341 provided on the side edge and an adjusting piece 342 provided on the arc guide rail 310 or the sliding seat 320. The adjusting piece 342 can adjust its position relative to the side edge and press 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 pieces 342 are provided and arranged at intervals. In the figure, the adjusting piece 342 can be positioned and movably arranged on the sliding seat 320, for example, it is screwed, and the adjusting piece 342 can be extended out or retracted into 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 piece 342 is adjusted independently. The sliding piece 341 can protect the side edge of the arc guide rail 310 to avoid wear while achieving uniform force, thereby ensuring the adjustment accuracy.

[0049] Reference Figure 5 To Attachment Figure 7 In the illustrated embodiment, the arc-shaped guide rail 310 has a front side facing the probe 100 and an opposite back side, the cross section of the sliding seat 320 is C-shaped and surrounds the arc-shaped guide rail 310, the C-shaped cross section of the sliding seat 320 accommodates the front side inside and the two ends of the C-shaped cross section opening pass through the side edge of the arc-shaped guide rail 310 and are close to the back side. Among them, the two ends of the C-shaped cross section opening of the sliding seat 320 can be set to be close to the back side of the arc-shaped guide rail 310 or to be set with a gap between the back side of the arc-shaped guide rail 310. When the two ends of the C-shaped cross section opening are close to the back side of the arc-shaped guide rail 310, they can be set to be stress-free close to provide a smooth sliding experience or mutually squeezed close to achieve mutual positioning. In the figure, the two ends of the C-shaped opening of the sliding seat 320 are movably provided with positioning pieces 321, and the sliding seat 320 cooperates with the back side of the arc-shaped guide rail 310 through the positioning pieces 321. Preferably, the positioning pieces 321 are arranged in pairs and correspond to the upper side edge and the lower side edge of the sliding seat 320 respectively. The positioning piece 321 is connected to the body of the sliding seat 320 through fasteners and adjusts the matching clearance between the sliding seat 320 and the arc guide rail 310, such as the mutual abutment or clearance matching mentioned above.

[0050] Further, the sliding seat 320 is provided with a cavity 322 facing the front and a plurality of sliding blocks 323 arranged at intervals around the cavity 322. The sliding block 323 is provided with a smooth sliding surface facing the front, and the sliding seat 320 cooperates with the front through the sliding surface of the sliding block 323. The sliding block 323 can improve the positioning accuracy of the two while maintaining the sliding seat 320 and the front sliding cooperation, avoiding the problem of reduced positioning accuracy or excessive cooperation resistance due to excessive contact area. In cooperation with it, the front part area of ​​the arc guide rail 310 is recessed compared to its side edge to form a recessed area, the recessed area corresponds to the cavity 322, and the sliding block 323 cooperates 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 posture of the probe 100. The position mark is located in the recessed area to avoid wear due to the relative movement of the sliding seat 320 and the arc guide rail 310.

[0051] The locking assembly also has a variety of configurations, for example, Figure 5As shown in , the locking assembly includes a sliding piece 341 and a locking rod 331 pressed against the sliding piece 341, wherein the locking assembly can share the same sliding piece 341 with the anti-backlash assembly 340. The locking rod 331 cooperates 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 achieving the locking or releasing of the sliding seat 320 and the arc guide rail 310. The locking assembly can also be implemented 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 achieve the clamping or releasing of the arc guide rail 310, thereby achieving the locking or releasing of the sliding seat 320 and the arc guide rail 310. The setting method of the locking assembly mentioned above can be implemented independently or in coordination.

[0052] In addition to motion guidance, the arc guide rail 310 can also provide a limit for the sliding seat 320. The two ends of one side edge of the arc guide rail 310 are raised to form a limit end 311, which abuts against the limit end 311 when the sliding seat 320 moves to the extreme position. In the direction of gravity, the limit end 311 and the anti-backlash component 340 can be arranged on the opposite side of the arc guide rail 310, for example, the limit end 311 is arranged on the side edge located below in the direction of gravity, and the anti-backlash component 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 311 and the anti-backlash component 340 can also be arranged on the same side of the arc guide rail 310.

[0053] The arc guide 310 and the base 301 are positioned by inserting and matching the positioning pins 312 and the positioning holes 313, and are connected to each other by fasteners. The positioning holes 313 are arranged in groups of two, and each group of positioning holes 313 is respectively located on the arc guide 310 and the base 301. There are multiple positioning pins 312 and they are respectively matched with a group of positioning holes 313. In one group of positioning holes 313, a positioning hole 313 located on the base 301 or on the arc guide 310 is a waist hole to allow the positioning pin 312 to fine-tune the relative position.

[0054] For details about the cooperation between the connection base 350 and the clamp 200, please refer to the attached Figure 5 In the illustrated embodiment, the connecting seat 350 includes clamping members 351 located on both radial sides of the clamp 200 and locking members 352 cooperating with the clamping members 351. The clamping members 351 form a clamping area, the clamp 200 is located in the clamping area, and the locking members 352 extend into the clamping area and position the clamp 200. Figure 5In the embodiment, the clamping member 351 is a 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, i.e., the clamping area. The locking member 352 and / or the clamping member 351 can cooperate with the outer peripheral surface of the clamp 200 to achieve the positioning of the clamp 200. Further, the clamp 200 is provided with an adjustable positioning ring 213, and the positioning ring 213 abuts against the clamping member 351 to pre-position the relative position of the clamp 200 relative to the connecting seat 350. In the direction of gravity, the positioning ring 213 cooperates with the upper end surface of the clamp 351.

[0055] In order to ensure that the front end of the probe 100 is in a certain spatial position compared to the holder 200, the holder 200 in this embodiment can achieve accurate positioning of the probe 100. Figure 8 In the illustrated embodiment, the holder 200 includes a housing, which includes a head 211 and a body 212 that are axially relatively movable. 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. The limiting portion 2122 is provided with a pressure channel 2123 that is connected to the inside of the probe 100. The limiting portion 2122 can be an annular structure arranged inside the body 212. In the specific implementation of the limiting portion 2122, referring to an embodiment, the body 212 is a multi-layer structure and includes:

[0056] The first cylinder 2124 is located at the outer layer and is 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 equipment;

[0057] The second cylinder 2125 is located inside the first cylinder 2124. The second cylinder 2125 includes an open end close to the clamping end 2127 and a closed end located inside the first cylinder 2124. The closed end forms a limiting portion 2122.

[0058] The third cylinder 2126 is located inside the first cylinder 2124 and is open at both ends, with one end abutting against the closed end of the second cylinder 2125 and the other end close to the pressure end 2128 of the first cylinder 2124 .

[0059] By embedding the second cylinder 2125 and the third cylinder 2126 in the first cylinder 2124, the positioning accuracy of the limit portion 2122 compared to the probe assembly can be guaranteed while reducing the processing difficulty, thereby ensuring the spatial position of the front end of the probe 100 compared to the probe assembly, thereby cooperating with the fixed-point rotating device 300 to realize the fixed-point adjustment of the front end of the probe 100.

[0060] In another embodiment, the air pressure source assembly 60 is used as a power source for aspirating and spitting out substances (cells, reagents, etc.). Fig. 9 In the illustrated embodiment, the air pressure source assembly includes a plunger pump 61, a solenoid valve 62, a three-way connector 63, a first one-way valve 64, a second one-way valve 65, an air filter 66, a muffler 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; and the three-way connector 63 has three connection ports.

[0061] The plug 68 is screwed into the inlet end 61a of the plunger pump, tightened and sealed, and the air intake and exhaust of the plunger pump are 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 muffler 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 connector 63 by a pipeline, and one of the other two ports of the three-way connector 63 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 then connected to the probe 100, so that the entire air pressure source assembly is connected to the probe 100.

[0062] The air filter 66 has a muffler 67 connected to the air inlet end to reduce the noise of the air path system of the entire operating cell. The air filter 66 has an exhaust end connected to the "IN" port of the first one-way valve 64, which prevents the gas in the entire air 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.

[0063] Before operation, the common end 62a and the normally open end 62b of the solenoid valve 62 are first connected, the normally closed end 62c is closed, and the plunger 61c first moves upward to the uppermost end, during which the residual gas in the plunger pump 61 is exhausted through the three-way 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 which the air is filtered by the air filter 66 and enters the plunger pump 61 through the first one-way valve 64 and the three-way joint 63. The initial stop position of the plunger 61c is set at the middle of the plunger pump, and it can move downward to provide negative air pressure, and can also move upward to provide positive air pressure.

[0064] In some embodiments, Fig.10 As shown, the plunger pump 61 provides positive or negative air pressure to the probe 100 in the form of pulses, and the plunger 61c moves intermittently within a single working stroke. When the plunger 61c moves intermittently upward, each movement distance 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 intermittently downward, each movement distance 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.

[0065] The origin O in the figure indicates that the plunger 61c is in the initial position, and the air pressure value in the plunger is close to the atmospheric pressure. Generally speaking, the pulse air pressure value is selected to be -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 practical needs.

[0066] The following uses cell extraction as an example to introduce the working principle of the air pressure source component:

[0067] The solenoid valve 62 is energized, the normally open end 62b is closed, and the normally closed end 62c is opened, so that the plunger pump 61 is connected to the probe 100, and the plunger 61c in the plunger pump 61 is controlled to move downward. At this time, the plunger pump 61 provides a negative air pressure pulse to the probe 100, and the probe 100 can suck the cells in the culture dish. If a single pulse cannot suck the cells into the probe 100, multiple pulses can be sent continuously until the cells are sucked in. During the process of the probe 100 sucking the cells, when the internal and external air pressures are balanced, the cells no longer move, so they can be kept in the probe 100.

[0068] After the cells are sucked in, if the plunger 61c has not moved to the limit position, it can continue to move to provide pulses to suck in more cells.

[0069] When the plunger 61c moves downward to the limit position, the normally closed end 62c of the solenoid valve 62 is closed and the normally open end 62b is connected, and the plunger 61c moves upward to reset. During the reset process, the gas in the plunger pump 61 is discharged through the three-way connector 63 and the second one-way valve 65. After the plunger 61c is reset, the normally open end 62b is closed and the normally closed end 62c is opened, and cells can be continuously sucked in. The reciprocating operation can be repeated to suck more cells at different sites into the probe 100 in batches.

[0070] After the cells are sucked in, the plunger 61c is reset first (or not reset), and the plunger 61c in the plunger pump 61 is controlled to move upward, and a positive air pressure pulse is provided to the probe 100 at this time, and the probe 100 spits out the cells into the culture dish. Since the positive air pressure is also provided in the form of pulses, the cells can be spewed out one by one until all of them are spewed out. The working process is opposite to the process of sucking cells, and will not be repeated here.

[0071] By using positive or negative air pressure pulses, at most one cell can be ejected or sucked in each operation. Providing a negative air pressure pulse may prevent the cell from entering the probe due to insufficient suction. By continuously outputting multiple pulses, the cell can eventually enter the probe 100. The ejection of cells is similar to this.

[0072] Since the probe has a large space inside, it can temporarily store more cells. The probe 100 can continuously suck in multiple cells and then spit out the cells at once, which significantly improves the operation efficiency. In addition, the pulse probe 100 can spit out cells one by one, so that multiple cells are distributed in different positions, eliminating the redistribution operation.

[0073] It can be seen that the solenoid valve 62 serves as a pressure balancing device for 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 gas path system connecting the probe 100 and the plunger pump 61 can maintain pressure balance, and the extracted cells can be completely preserved in the probe 100. The plunger 61c can then 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 plunger pump 61 performs a reset operation, which does not affect the cells preserved in the probe 100.

[0074] The microscopic imaging assembly includes a light source 23, an optical microscope objective lens 21, and an image detection unit 22 (such as a CCD camera). The light source is generally arranged directly above the stage 10, and the optical microscope objective lens 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, the solenoid valve 62, the motor in the mobile platform 50, and the motor in the stage 10 in the air pressure source assembly 60 are controlled by the 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.

[0075] Other details of the single-cell manipulation device can be implemented in combination with existing technologies and will not be described in detail here.

[0076] The technical features of the above-mentioned embodiments can be combined arbitrarily. To make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification. When the technical features in different embodiments are embodied in the same figure, it can be regarded that the figure also discloses the combination examples of the various embodiments involved.

[0077] The above-mentioned embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the patent application. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application.

Claims

1. A single cell manipulation device, characterized in that: include: A stage for fixing the container carrying the sample; A probe assembly comprises a probe and a holder for clamping the probe, wherein the holder comprises a shell, wherein the shell comprises a head and a body which are relatively movable in the axial direction, wherein the body is a multi-layer structure and comprises a first cylinder, a second cylinder and a third cylinder; the first cylinder is located in the outer layer and is open at both ends, one end is a clamping end extending into the head, and the other end is a pressure end connected to a peripheral device; the second cylinder is located inside the first cylinder, wherein the second cylinder comprises an open end close to the clamping end and a closed end located inside the first cylinder, wherein the closed end forms a limiting portion, and a pressure channel connected to the inside of the probe is provided on the limiting portion; the third cylinder is located inside the first cylinder and is open at both ends, wherein one end abuts against the closed end of the second cylinder, and the other end is close to the pressure end of the first cylinder; the probe is an elongated hollow structure, having a front end and a rear end opposite to each other, wherein the front end of the probe extends to the container when in use, and the rear end of the probe passes through the head and extends into the body and is positioned at the limiting portion; A control assembly, comprising a fixed-point rotating device and a mobile platform for moving the probe assembly in space to adjust the position of the probe, wherein the fixed-point rotating device comprises a mutually cooperating arc-shaped guide rail, a sliding seat, and a locking assembly for locking the relative positions of the two, wherein 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 is linked to the mobile 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 the three-dimensional space; A microscopic imaging component, providing a window for positioning the probe; An air pressure source component is connected to the rear end of the probe via the clamp to provide positive or negative air pressure for the probe.

2. The single cell manipulation device according to claim 1, characterized in that: 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 of the arc-shaped guide rail; or The extension path of the arc-shaped guide rail is located on the conical surface of a vertebral body, and the front end of the probe is located at the apex position of the vertebral body.

3. The single cell manipulation device according to claim 1, characterized in that: 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 two sides of the arc-shaped guide rail in the extending direction of the probe.

4. The single cell manipulation device according to claim 3, characterized in that: A gap elimination component is also provided between the arc guide rail and the sliding seat. The gap elimination component includes a sliding piece arranged on the side edge and an adjusting member arranged on the arc 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, characterized in that: The sliding sheet and the side edge extend in the same direction, and in the extending direction of the sliding sheet, a plurality of the adjusting members are provided and arranged at intervals.

6. The single cell manipulation device according to claim 1, characterized in that: The arc guide rail has a front side facing the probe and an opposite back side. The sliding seat has a C-shaped cross section and surrounds the arc guide rail. The C-shaped interior of the sliding seat accommodates the front side and both ends of the C-shaped opening pass through the side edges of the arc guide rail and are close to the back side.

7. The single cell manipulation device according to claim 6, characterized in that: The sliding seat is provided with a cavity toward the front side and a plurality of sliding blocks arranged at intervals are provided around the cavity, and the sliding seat cooperates with the front side through the sliding blocks; A position mark for marking the position of the probe is provided on the front side.

8. The single cell manipulation device according to claim 1, characterized in that: The connecting seat includes a clamping member located radially of the clamp and a locking member matched with the clamping member, the clamping member forms a clamping area, the clamp is located in the clamping area, and the locking member extends into the clamping area and positions the clamp.

9. The single cell manipulation device according to claim 8, characterized in that: The clamp is provided with an adjustable positioning ring, and the positioning ring abuts against the clamping member to pre-position the relative position of the clamp relative to the connecting seat.

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