CN115572670A

CN115572670A - Cell micromanipulation device and multi-cell management method

Info

Publication number: CN115572670A
Application number: CN202211480914.3A
Authority: CN
Inventors: 陈方鑫; 郭汝海; 韦俊杰; 毕海
Original assignee: Ji Hua Laboratory
Current assignee: Ji Hua Laboratory
Priority date: 2022-11-24
Filing date: 2022-11-24
Publication date: 2023-01-06
Anticipated expiration: 2042-11-24
Also published as: CN115572670B

Abstract

The invention relates to the technical field of micromanipulation, and discloses a cell micromanipulation device and a multi-cell management method. The cell micromanipulation apparatus includes: the mobile control mechanism comprises a visual component, an operating platform and a plurality of mobile structures, wherein the operating platform is provided with a culture dish, and the mobile structures are arranged around the operating platform; the visual component is arranged on one side of the culture dish; a micro-manipulation mechanism including a storage needle for storing a cell culture solution, an adsorption needle for moving cells, a storage needle for storing treated cells, and an injection needle for injecting cells, the micro-manipulation mechanism being provided on the moving structure; and the servo driving mechanism is used for outputting the specified pressure. According to the invention, a small amount of cells are controlled by the micro-operation mechanism once, and a large amount of cells can be managed by repeatedly using the micro-operation mechanism for many times, so that the automatic management of multiple cells is realized.

Description

Cell micromanipulation device and multi-cell management method

Technical Field

The invention relates to the technical field of micromanipulation, in particular to a cell micromanipulation device and a multi-cell management method.

Background

A micromanipulation device, also known as a micromanipulation robot, is a device used to microscopically perform microsurgical operations and injections of cells. The device is controlled by a mechanical structure or a circuit, and a tiny tool is used for grasping, cutting, extruding, injecting, retracting and the like under the guidance of a microscope. The micromanipulation device can be used for nuclear transfer, gene injection, embryo cutting and other operations, and can also be used for separating single cells and operating or managing single or multiple cells.

In the prior art, a chinese patent application No. 201110359226.7 entitled "a method for repositioning cells in batches by a micro-operation robot system" discloses a method for identifying and positioning relative positions of multiple cells, and the method establishes a corresponding relationship for the multiple cells simultaneously by using visual information. The method is applied to a micro-operation robot system, and has convenience in batch operation. The Chinese invention patent with the application number of '201910449314.2' and the name of 'a micro-channel-based robotic somatic cell nuclear transfer operation method' introduces the micro-channel for storing cells and introduces parallel double needles for cell enucleation and nuclear injection respectively. Compared with the traditional somatic cell nuclear transfer process, the method saves time, has the same survival rate and success rate as manual somatic cell nuclear transfer, and effectively improves the efficiency of somatic cell nuclear transfer. The method can realize batch management or operation of cells, but the first patent completely depends on visual information, and the essence of the method is to group and classify the cells in a visual field by using a visual algorithm, so that the management of the cells is realized at a program level, namely, only the cells in the visual field can be effectively managed, the cells outside the visual field of a microscope can be lost, and no method is available for realizing effective management. The second patent uses a micro flow channel to confine cells at a specific position, and this method, although the position of the cells is ordered, achieves the effect of managing cell groups, requires that the cells are put into the micro flow channel in sequence and arranged orderly in the early stage, and the process is complex and has a large workload, and the position of the cells may drift away to be controlled along with the disturbance of the fluid during the operation.

Disclosure of Invention

The invention mainly aims to provide a cell micromanipulation device and a multi-cell management method, aiming at solving the technical problem that cells leaving the visual field cannot be managed.

To achieve the above object, the present invention provides a cell micromanipulation apparatus comprising: the mobile control mechanism comprises a visual component, an operating platform and a plurality of mobile structures, wherein a culture dish is arranged on the operating platform, and the mobile structures are arranged around the operating platform; the visual component is arranged on one side of the culture dish; a micro-manipulation mechanism provided on the moving structure, the micro-manipulation mechanism including a storage needle for storing a cell culture solution, an adsorption needle for moving cells, a storage needle for storing processed cells, and an injection needle for injecting cells; the servo driving mechanism is used for outputting driving force, and the micro-operation mechanism is connected with the output end of the servo driving mechanism.

Optionally, the servo driving mechanism includes a three-way valve, a fourth linear motion assembly and an air pressure sensor, the fourth linear motion assembly includes a servo driving module and a piston type pneumatic pump, the servo driving module is connected with the piston type pneumatic pump, the piston type pneumatic pump is connected with the micro-operation mechanism, the piston type pneumatic pump is used for driving the micro-operation mechanism, the air pressure sensor, the piston type pneumatic pump and the micro-operation mechanism are respectively communicated with three channels of the three-way valve, and the air pressure sensor is electrically connected with the fourth linear motion assembly.

Optionally, the mobile structure includes a first linear motion assembly, a second linear motion assembly, a third linear motion assembly and an electric rotary table, the second linear motion assembly is arranged on the third linear motion assembly, the first linear motion assembly is arranged on the second linear motion assembly, the electric rotary table is arranged on the first linear motion assembly, and the micro-operation mechanism is arranged on the electric rotary table.

Optionally, the operating platform includes a driving motor, a first tooth-shaped member, a base and a second tooth-shaped member, the first tooth-shaped member is rotatably disposed on the base, the second tooth-shaped member is engaged with the first tooth-shaped member, the second tooth-shaped member is rotatably disposed on the base, a mounting hole corresponding to the shape of the culture dish is disposed on the second tooth-shaped member, and the culture dish is mounted in cooperation with the mounting hole.

Optionally, the visual component includes a first camera, a lens and a light source, the light source is disposed directly above the operating platform, the lens is disposed on a surface of the operating platform deviating from the light source, and the first camera is connected to the lens.

Optionally, the visual component includes a first camera, a second camera, a lens, a coaxial corner module and a light source, the light source is disposed over the operating platform, the lens is movably disposed on a surface of the operating platform facing away from the light source, the coaxial corner module is connected with the lens, the first camera is connected with the coaxial corner module, and the second camera is connected with the coaxial corner module.

Optionally, the visual assembly further comprises a lifting structure, the lifting structure is fixedly arranged below the operating platform, the visual assembly is arranged on the lifting structure, and the lifting structure is used for adjusting the distance between the lens and the culture dish.

The invention also provides a multi-cell management method applied to the cell micromanipulation device, and the multi-cell management method comprises the following steps:

s1: controlling the moving structure to move the storage needle into the culture dish, and controlling the servo driving mechanism to drive the storage needle to suck cell culture solution containing cells;

s2: controlling the moving structure to move the storage needles to the appointed positions in the culture dish, and controlling the servo driving mechanism to drive the storage needles to release cells in the culture dish one by one;

s3: the visual assembly is controlled to acquire the position information of the cell to be operated in the visual field, the moving structure is controlled to move the adsorption needle to the cell to be operated, the servo driving mechanism is controlled to drive the adsorption needle to adsorb the cell, and the cell is moved to a specified position;

s4: controlling the moving structure to move the injection needle until the needle head of the injection needle punctures the cell membrane of the cell, and controlling the servo driving mechanism to drive the injection needle to inject solution into the cell;

s5: the vision assembly is controlled to acquire the position information of the cell injected with the solution, the moving structure is controlled to move the accommodating needle to the position of the cell injected with the solution, and the servo driving mechanism is controlled to drive the accommodating needle to recover the cell injected with the solution.

Optionally, in step S2, the minimum pressure P required by the storage needle when the cells are released is ₁ The calculation formula of (2) is as follows:

;

wherein gamma is the fluid surface tension coefficient, theta is the contact angle, d is the diameter of the needle tube, rho is the liquid density, V is the fluid velocity, L is the fluid length, and lambda is the pipeline friction coefficient.

Optionally, in step S5, the minimum recovery pressure P required for accommodating the needle is set ₂ The calculation formula of (c) is:

;

wherein gamma is the fluid surface tension coefficient, theta is the contact angle, d is the diameter of the needle tube, rho is the liquid density, V is the fluid velocity, L is the fluid length, xi is the loss coefficient, and lambda is the pipeline friction coefficient.

The cell micromanipulation device is provided with a micromanipulation mechanism with a pressure servo function, a storage needle, a containing needle, an adsorption needle and an injection needle in the micromanipulation mechanism are all modified based on a pneumatic microinjector, and a servo driving mechanism outputs air pressure to control the action of the micromanipulation mechanism. Wherein the adsorption needle and the injection needle are provided with needle heads for operating cells, and the storage needle are provided with cavities for accommodating cells and cell culture solution. The storage needle is used for storing cells to be operated, so that cell searching time is reduced; the injection needle and the adsorption needle are used for performing operations such as injection, movement and the like on cells; the storage needle is used for collecting cells injected with a solution, and management confusion caused by mixing of the operated cells and the cells to be operated is avoided. In the traditional method, the multi-cell operation mostly depends on a visual algorithm, and the cells can not be managed and controlled after leaving the visual field and losing control. Cells outside the field of view will not be managed and controlled. In this scheme, when the cell quantity is more, when the field of vision can't hold, all unoperated cells are deposited to the accessible memory needle, accomodate the needle and accomodate all cells after the operation, release the cell again when needing to manage the cell, release the cell operation by adsorbing the needle. An operator can stably manage and control single or multiple cells according to the steps by using the cell micromanipulation device, and can automatically manage the cells in a large batch by repeating the steps, so that the problem that the cells cannot be managed in a visual field is solved, and the real full-automatic multi-cell management is realized.

Drawings

One or more embodiments are illustrated in drawings corresponding to, and not limiting to, the embodiments, in which elements having the same reference number designation may be represented as similar elements, unless specifically noted, the drawings in the figures are not to scale.

FIG. 1 is a schematic view of the overall structure of one embodiment of a cell micromanipulation apparatus according to the present invention;

FIG. 2 is an enlarged view of a portion of FIG. 1 at A;

FIG. 3 is a schematic left side view of a cell micromanipulation apparatus according to an embodiment of the present invention;

FIG. 4 is a schematic view showing the overall structure of a servo driving mechanism according to an embodiment of the cell micromanipulation apparatus of the present invention;

FIG. 5 is a schematic flow chart of a method for managing multiple cells in a cell micromanipulation apparatus according to the present invention;

FIG. 6 is a schematic view of cell collection and release according to an embodiment of the present invention;

FIG. 7 is a microscopic view of a receiving needle according to an embodiment of the present invention;

FIG. 8 is a microscopic view of an initial stage of releasing cells from a receiving needle according to an embodiment of the present invention;

FIG. 9 is a microscopic view of an intermediate stage of releasing cells from a receiving needle according to an embodiment of the present invention;

FIG. 10 is a microscopic view of the end stage of receiving a needle releasing cells according to an embodiment of the present invention.

Wherein, 1, a cell micromanipulation device; 2. a movement control mechanism; 21. a visual component; 211. a first camera; 212. a second camera; 213. a lens; 214. a light source; 215. a coaxial corner module; 216. a lifting structure; 22. an operating platform; 221. a culture dish; 222. a drive motor; 223. a first tooth-shaped member; 224. a base; 225. a second toothed member; 2251. mounting holes; 23. a moving structure; 231. a first linear motion assembly; 232. a second linear motion assembly; 233. a third linear motion assembly; 234. an electric rotating table; 3. a micro-operation mechanism; 31. a storage needle; 32. an adsorption needle; 33. a receiving needle; 34. an injection needle; 4. a servo drive mechanism; 41. a three-way valve; 42. a fourth linear motion assembly; 421. a servo drive module; 422. a piston type pneumatic pump; 4221. a piston; 43. an air pressure sensor; 5. a cell; 6. and a processing module.

Detailed Description

In order to facilitate an understanding of the invention, reference will now be made in detail to the present embodiments of the invention, examples of which are illustrated in the accompanying drawings. It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may be present. The terms "vertical," "horizontal," "left," "right," "inner," "outer," and the like as used herein are for purposes of description only. In the description of the present invention, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating relative importance or as implicitly indicating the number of technical features indicated. Thus, unless otherwise specified, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature; "plurality" means two or more. The terms "comprises" and any variations thereof, are intended to cover a non-exclusive inclusion, which may have the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or combinations thereof.

Furthermore, unless expressly stated or limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly and may include, for example, fixed connections, removable connections, and integral connections; can be mechanically or electrically connected; either directly or indirectly through intervening media, or through both elements. All technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Furthermore, the technical features mentioned in the different embodiments of the invention described below can be combined with each other as long as they do not conflict with each other.

As shown in fig. 1 to 3, the present invention provides a cell micromanipulation apparatus 1, the cell micromanipulation apparatus 1 including: the movement control mechanism 2 comprises a vision assembly 21, an operating platform 22 and a plurality of moving structures 23, wherein a culture dish 221 is arranged on the operating platform 22, and the plurality of moving structures 23 are arranged around the operating platform 22; the vision assembly 21 is arranged on one side of the culture dish 221; a micromanipulation mechanism 3, the micromanipulation mechanism 3 being provided on the movement mechanism 23, the micromanipulation mechanism 3 including a storage needle 31 for storing a cell culture solution, an adsorption needle 32 for moving a cell, a housing needle 33 for housing a processed cell, and an injection needle 34 for injecting a cell; the servo driving mechanism 4 is used for outputting driving force, and the micro-operation mechanism 3 is connected with the output end of the servo driving mechanism 4.

In the present invention, a micromanipulator 3 having a pressure servo function is incorporated in a cell micromanipulator 1, a storage needle 31, a storage needle 33, an adsorption needle 32, and an injection needle 34 in the micromanipulator 3 are modified based on a pneumatic microinjector, and a servo driver 4 outputs air pressure to control the operation of the micromanipulator 3. The suction needle 32 and the injection needle 34 have needle heads for manipulating cells, and the storage needle 33 and the storage needle 31 have cavities for accommodating cells. The storage needle 31 is used for storing cells to be operated, so that the cell searching time is reduced; the injection needle 34 and the adsorption needle 32 are used for injecting and moving cells; the storage needle 33 is used for collecting the operated cells, and avoids management confusion caused by mixing the operated cells and the cells to be operated. In the traditional method, the multi-cell operation mostly depends on a visual algorithm, and the cells can not be managed and controlled after leaving the visual field and losing control. Cells outside the field of view will not be managed and controlled. In this embodiment, when the number of cells is large and the field of view cannot be stored, all the cells that have not been operated can be stored in the storage needle 31, all the operated cells can be stored in the storage needle 33, and the cells can be released and operated by the adsorption needle 32 and the injection needle 34 when the cells need to be managed. An operator can stably manage and control single or multiple cells by using the cell micromanipulation device 1 according to the steps, and can automatically manage a large number of cells by repeating the steps, so that the problem that the cells cannot be managed in a visual field is solved, and the full-automatic multi-cell management in the true sense is realized.

As shown in fig. 1 and 4, the servo driving mechanism 4 includes a three-way valve 41, a fourth linear motion assembly 42 and a pneumatic pressure sensor 43, the fourth linear motion assembly 42 includes a servo driving module 421 and a piston type pneumatic pump 422, the servo driving module 421 is connected to the piston type pneumatic pump 422, the piston type pneumatic pump 422 is pneumatically connected to the micro-operation mechanism 3, the specified pneumatic pressure can be output by pushing the piston type pneumatic pump 422 to control the storage needle 31 and the storage needle 33 to absorb or release cells, the absorption needle 32 absorbs or releases cells, and the injection needle 34 injects a solution into the cells. Each micro-actuator 3 is connected to a servo drive 4. The air pressure sensor 43, the piston type air pressure pump 422 and the micro-operation mechanism 3 are respectively communicated with three channels of the three-way valve 41, and the air pressure sensor 43 is electrically connected with the fourth linear motion assembly 42. The servo driving module 421 includes parts such as a servo motor, a coupling, a transmission platform, etc., and can be electrically connected to the servo motor through an external industrial computer or a programmable controller to control the rotation speed and the output torque of the servo motor, so as to push the piston 4221 in the piston type pneumatic pump 422 to move forward or backward. The air pressure sensor 43, which is in pipe communication with the three-way valve 41, is used for detecting real-time air pressure in the pipe, and then the servo controller controls the servo driving module 421 to push the piston 4221 of the piston type air pressure pump 422, so as to change the air pressure in the pipe, so that the real-time air pressure approaches to the specified air pressure, and the micro-operation mechanism 3 can be accurately controlled to operate the cells on the premise of not damaging the cells.

As shown in fig. 1 and 2, in the present embodiment, the moving structure 23 includes a first linear motion assembly 231, a second linear motion assembly 232, a third linear motion assembly 233 and an electric rotating table 234, the second linear motion assembly 232 is disposed on the third linear motion assembly 233, the first linear motion assembly 231 is disposed on the second linear motion assembly 232, the electric rotating table 234 is disposed on the first linear motion assembly 231, and the micro-operation mechanism 3 is disposed on the electric rotating table 234. The first linear motion assembly 231, the second linear motion assembly 232 and the third motion assembly are common linear motion modules formed by components such as a motor, a ball screw and a guide rail, the electric rotating platform 234 is a single-degree-of-freedom rotating bearing platform formed by the motor and transmission parts, and the micro-operation mechanism 3 is arranged on the bearing platform in the electric rotating platform 234. The micro-manipulator 3 moves in three directions, i.e., x-axis, y-axis and z-axis, based on the moving structure 23 formed by assembling the first linear motion module 231, the second linear motion module 232 and the third linear motion module, and moves the micro-manipulator 3 to any region in the culture dish 221 to manipulate cells by using the electric rotating table 234 capable of rotating in one degree of freedom.

As shown in fig. 1 and fig. 2, in the present embodiment, the operation platform 22 includes a driving motor 222, a first tooth member 223, a base 224, and a second tooth member 225, the first tooth member 223 is rotatably disposed on the base 224, the second tooth member 225 is engaged with the first tooth member 223, the second tooth member 225 is rotatably disposed on the base 224, a mounting hole 2251 corresponding to the shape of the culture dish 221 is disposed on the second tooth member 225, and the culture dish 221 is mounted in cooperation with the mounting hole 2251. When the driving motor 222 works, the placing direction of the culture dish 221 can be rotated, the first tooth-shaped member 223 and the second tooth-shaped member 225 are in gear transmission, and the micro-operation mechanisms 3 can move to any area in the culture dish 221 by matching with the movement control mechanism 2, and the plurality of micro-operation mechanisms 3 cannot be interfered with each other.

As shown in fig. 1 and fig. 3, in the present embodiment, the visual component 21 includes a first camera 211, a lens 213 and a light source 214, the light source 214 is disposed right above the operation platform 22, the lens 213 is disposed on a surface of the operation platform 22 away from the light source 214, and the first camera 211 is connected to the lens 213. The vision assembly 21 is primarily arranged in a back-lit light source arrangement for viewing detailed features of the object to be observed.

As shown in fig. 1 and fig. 3, in the present embodiment, the visual component 21 includes a first camera 211, a second camera 212, a lens 213, a coaxial rotation module 215 and a light source 214, the light source 214 is disposed directly above the operation platform 22, the lens 213 is movably disposed on a surface of the operation platform 22 away from the light source 214, the coaxial rotation module 215 is connected to the lens 213, the first camera 211 is connected to the coaxial rotation module 215, and the second camera 212 is connected to the coaxial rotation module 215. The first camera 211 and the second camera 212 are laboratory biomicroscopic cameras, wherein the second camera 212 is a small-magnification camera for a large field of view for searching for cells to be manipulated in the field of view; the first camera 211 is a large-power camera for observing detailed features of the cells. The coaxial rotation angle module 215 distributes the light path to the two cameras, so as to achieve the purpose that the two cameras work simultaneously.

As shown in fig. 1 and fig. 3, in the present embodiment, the vision assembly 21 further includes a lifting structure 216, the lifting structure 216 is fixedly disposed below the operation platform 22, the vision assembly 21 is disposed on the lifting structure 216, and the lifting structure 216 is used for adjusting a distance between the lens 213 and the culture dish 221. Elevation structure 216 is shear type vertical lift, and visual component 21 installs on elevation structure 216's platform, and elevation structure 216 drives visual component 21 vertical movement from top to bottom, and elevation structure 216 can change the distance between camera lens 213 and the culture dish 221 for the imaging distance between adjustment camera and the culture dish 221.

As shown in fig. 4 to 10, the present invention further provides a multi-cell management method applied to the cell micromanipulation apparatus 1, the multi-cell management method including the steps of:

s1: the movement structure is controlled to move the storage needle 31 into the culture dish 221, and the servo driving mechanism 4 is controlled to drive the storage needle 31 to suck cell culture solution containing cells;

in this step, the storage needle 31 stores all the cells 5 to be managed together in advance, thereby facilitating the uniform management.

S2: controlling the moving mechanism 23 to move the storage needle 31 to a specified position in the culture dish 221, and controlling the servo driving mechanism 4 to drive the storage needle 31 to release the cells 5 in the culture dish 221 one by one;

in this step, the servo driving mechanism 4 controls the real-time air pressure through the three-way valve 41 and the air pressure sensor 43, so as to prevent the servo driving mechanism 4 from providing too much pressure into the storage needle 31 to damage the cell 5 when the storage needle 31 releases the cell 5, or prevent the servo driving mechanism 4 from providing too little pressure to cause the cell 5 to be stuck in the storage needle 31.

S3: the control vision assembly 21 acquires the position information of the cell 5 to be operated in the visual field, the control moving structure 23 moves the adsorption needle 32 to the cell 5 to be operated, the control servo driving mechanism 4 drives the adsorption needle 32 to adsorb the cell 5, and the cell 5 is moved to a specified position;

in this step, the cells 5 are moved to a designated position by the suction needle 32 guided by the vision unit 21, and are laid down for further operation.

S4: controlling the moving structure 23 to move the injection needle 34 until the needle head of the injection needle 34 punctures the cell membrane of the cell 5, and controlling the servo driving mechanism 4 to drive the injection needle 34 to inject the solution into the cell 5;

in this step, the injection needle 34 can perform operations such as saline, cell marking, and cell nucleus injection on the cells 5 according to the experiment requirements.

S5: the control vision assembly 21 acquires the position information of the cell 5 injected with the solution, the control moving mechanism 23 moves the storage needle 33 to the position where the cell 5 is operated, and the control servo driving mechanism 4 drives the storage needle 33 to suck the cell 5 injected with the solution.

The steps S2 to S5 are repeated until all the cells 5 in the storage needle 31 are released and recovered into the storage needle 33. The cell micromanipulation device performs stable control on a small number of cells in each round of S2 to S5 steps, and repeats the S2 to S5 steps for multiple times until the operation on all cells in the cell culture solution in the storage needle 31 is completed, so that automatic management on a large number of cells is realized.

The cell micromanipulation device 1 is internally provided with a processing module 6, the processing module 6 can receive a real-time image obtained by shooting the culture dish 221 by the first camera 211 or the second camera 212 in the vision component 21, the processing module 6 processes the image of the obtained coordinates of the cell 5, the processing module 6 controls corresponding motors in the first linear motion component 231, the second linear motion component 232, the third linear motion component 233 or the electric rotating platform 234 in the moving structure 23 to work after the coordinates of the cell 5 are obtained, each motor outputs torque to convey the micromanipulation mechanism 3 to a specified position coordinate, and the guidance of the micromanipulation mechanism 3 is completed. In the whole process from S1 to S5, the processing module 6 includes a servo driver, which can control the servo driving mechanism 4, the processing module 6 sends out a signal according to a specified action to control a servo motor in the servo driving mechanism 4 to drive a piston type pneumatic pump 422 to output a specified driving force, which is used for controlling the storage needle 31 to suck the cell 5 culture solution or discharge the cells 5 one by one, and controlling the suction needle 32 to suck or discharge the cells 5, and also controlling the injection needle 34 to inject the solution, controlling the containing needle 33 to suck the cells 5, and after the whole process is finished, the servo driving mechanism 4 and the movement control mechanism 2 are reset. After the treatment of the cells 5 is completed, the receiving needle 33 releases the treated cells 5, so as to rapidly provide the treated cells 5 in the subsequent experiments, as shown in fig. 8 to 10, which are images of each stage of releasing the cells 5 by the receiving needle 33.

Optionally, in step S2, the minimum pressure P required to store needle 31 when cell 5 is released is ₁ The calculation formula of (c) is:

；

wherein gamma is the fluid surface tension coefficient, theta is the contact angle, d is the diameter of the needle tube, rho is the liquid density, V is the fluid velocity, L is the fluid length, and lambda is the pipeline friction coefficient. The minimum recovery pressure P is provided by the servo drive 4 connected to the storage needle 31 ₁ 。

Optionally, in step S5, the minimum recovery pressure P required to house the needle 33 is set ₂ The calculation formula of (c) is:

；

wherein gamma is the fluid surface tension coefficient, theta is the contact angle, d is the diameter of the needle tube, rho is the liquid density, V is the fluid velocity, L is the fluid length, xi is the loss coefficient, and lambda is the pipeline friction coefficient. The minimum recovery pressure P is provided by the servo-drive mechanism 4 connected to the receiving needle 33 ₂ 。

The above examples are only intended to illustrate the technical solution of the present invention, and not to limit it; within the idea of the invention, also technical features in the above embodiments or in different embodiments may be combined, steps may be implemented in any order, and there are many other variations of the different aspects of the invention as described above, which are not provided in detail for the sake of brevity; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and these modifications or substitutions do not depart from the spirit of the corresponding technical solutions of the embodiments of the present invention.

Claims

1. A cell micromanipulation apparatus, comprising:

the mobile control mechanism comprises a visual component, an operating platform and a plurality of mobile structures, wherein a culture dish is arranged on the operating platform, and the mobile structures are arranged around the operating platform; the visual component is arranged on one side of the culture dish;

a micro-manipulation mechanism provided on the moving structure, the micro-manipulation mechanism including a storage needle for storing a cell culture solution, an adsorption needle for moving cells, a storage needle for storing processed cells, and an injection needle for injecting cells;

the servo driving mechanism is used for outputting driving force, and the micro-operation mechanism is connected with the output end of the servo driving mechanism.

2. The cell micromanipulation apparatus according to claim 1, wherein the servo driving mechanism comprises a three-way valve, a fourth linear motion assembly and a gas pressure sensor, the fourth linear motion assembly comprises a servo driving module and a piston type gas pressure pump, the servo driving module is connected with the piston type gas pressure pump, the piston type gas pressure pump is connected with the micromanipulation mechanism, the piston type gas pressure pump is used for driving the micromanipulation mechanism, the gas pressure sensor, the piston type gas pressure pump and the micromanipulation mechanism are respectively communicated with three channels of the three-way valve, and the gas pressure sensor is electrically connected with the fourth linear motion assembly.

3. The cell micromanipulation apparatus of claim 2, wherein the moving structure comprises a first linear motion member, a second linear motion member, a third linear motion member, and a motor-driven rotation stage, wherein the second linear motion member is disposed on the third linear motion member, the first linear motion member is disposed on the second linear motion member, the motor-driven rotation stage is disposed on the first linear motion member, and the micromanipulation mechanism is disposed on the motor-driven rotation stage.

4. The cell micromanipulation apparatus of claim 1, wherein the manipulation platform comprises a driving motor, a first tooth-shaped member, a base and a second tooth-shaped member, the first tooth-shaped member is rotatably disposed on the base, the second tooth-shaped member is engaged with the first tooth-shaped member, the second tooth-shaped member is rotatably disposed on the base, a mounting hole corresponding to the shape of the culture dish is disposed on the second tooth-shaped member, and the culture dish is mounted in cooperation with the mounting hole.

5. The cell micromanipulation apparatus of claim 1, wherein the vision assembly comprises a first camera, a lens and a light source, the light source is disposed right above the operation platform, the lens is disposed on a surface of the operation platform facing away from the light source, and the first camera is connected to the lens.

6. The cell micromanipulation apparatus of claim 1, wherein the vision assembly comprises a first camera, a second camera, a lens, a coaxial rotation module and a light source, the light source is disposed directly above the operation platform, the lens is movably disposed on a surface of the operation platform facing away from the light source, the coaxial rotation module is connected to the lens, the first camera is connected to the coaxial rotation module, and the second camera is connected to the coaxial rotation module.

7. The cell micromanipulation apparatus of claim 6, wherein the vision assembly further comprises a lifting structure, the lifting structure is fixedly arranged below the operation platform, the vision assembly is arranged on the lifting structure, and the lifting structure is used for adjusting the distance between the lens and the culture dish.

8. A multi-cell management method applied to the cell micromanipulation apparatus according to any one of claims 1 to 7, wherein the multi-cell management method comprises the steps of:

s2: controlling the moving structure to move the storage needles to a specified position in the culture dish, and controlling the servo driving mechanism to drive the storage needles to release cells in the culture dish one by one;

9. The method for multi-cell management according to claim 8, wherein in the step S2, the minimum pressure P required by the storage needle when the cell is released is P ₁ The calculation formula of (2) is as follows:

;

wherein gamma is the surface tension coefficient of the fluid, theta is the contact angle, d is the diameter of the needle tube, rho is the density of the liquid, V is the velocity of the fluid, L is the length of the fluid, and lambda is the friction coefficient of the pipeline.

10. The method for managing multiple cells according to claim 8, wherein in step S5, the minimum recovery pressure P required for the needle to be housed is set to ₂ The calculation formula of (2) is as follows:

;