CN113583847A - Cell microinjection device and robust impedance control method thereof - Google Patents
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- 238000000520 microinjection Methods 0.000 title claims abstract description 69
- 238000000034 method Methods 0.000 title claims abstract description 26
- 230000007246 mechanism Effects 0.000 claims abstract description 56
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- 238000006073 displacement reaction Methods 0.000 claims description 12
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- 238000007789 sealing Methods 0.000 claims description 2
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- 230000006378 damage Effects 0.000 abstract description 5
- 210000004027 cell Anatomy 0.000 description 40
- 239000011148 porous material Substances 0.000 description 12
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- 239000007924 injection Substances 0.000 description 9
- 230000033001 locomotion Effects 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 5
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- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 210000000287 oocyte Anatomy 0.000 description 2
- 238000005299 abrasion Methods 0.000 description 1
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- 238000005452 bending Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
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- 238000010367 cloning Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 230000002900 effect on cell Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
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- 210000003463 organelle Anatomy 0.000 description 1
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- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
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Abstract
The invention discloses a cell microinjection device, which comprises a connecting rod, an encAN _ SNsulator connected with the connecting rod, a piezoelectric driver fixed in the encAN _ SNsulator, a microinjection needle, an injector and a hub type compliant mechanism, wherein two ends of the injector are respectively connected with the encAN _ SNsulator and the hub type compliant mechanism; the hub type compliant mechanism is provided with a reed type compliant unit for transmitting axial vibration to the micro-injection needle. The invention can reduce the transverse vibration of the needle head in the microinjection device and reduce the damage of cells in the process of cell puncture.
Description
Technical Field
The invention relates to the technical field of precise control and the field of biomedical engineering, in particular to a cell microinjection device and a robust impedance control method thereof.
Background
The microinjection technology is a technology for controlling a micromanipulator to operate cells or early embryos by operating the micromanipulator under a high-power inverted microscope, and is widely applied to the fields of drug development, in-vitro fertilization, biological cloning, transgenosis and the like. The core of microinjection technology is efficient, low-damage cell or early embryo puncture. Due to the low success rate and efficiency of manual puncture, the puncture process begins to develop towards automation. The automatic puncture technology widely adopts a piezoelectric driver to drive a micro-injection needle to complete the puncture process. However, studies have shown that piezoelectric signals induce needle tip vibration, resulting in cell deformation, where lateral vibration has a greater effect on cell deformation than axial vibration. Excessive cell deformation can stress and change the relative positions of organelles inside the cell, and further damage the cell.
At present, structurally, a compliant guide mechanism can effectively inhibit lateral vibration of a microinjection needle tip. The flexible mechanism mainly depends on the deformation of a flexible component in the mechanism to realize the main movement and function of the mechanism, and is widely applied to the field of precision transmission due to the advantages of high resolution, high precision and the like. The flexible guide mechanism is a flexible mechanism for guiding movement and has a guide effect on input displacement, so that the transverse vibration of the needle tip can be effectively inhibited, and further, the damage of cells is reduced. Therefore, the degree of cell damage in the current automatic puncturing technology depends greatly on the structural design of the compliant mechanism. The compliant mechanisms can be divided into a centralized compliant mechanism and a distributed compliant mechanism, and the difference is that the stress of the compliant section of the centralized compliant mechanism is distributed in a concentrated manner, and the stress of the compliant section of the distributed compliant mechanism is distributed in a dispersed manner; the compliant mechanism can be further divided into a space compliant mechanism, a plane compliant mechanism and the like according to the structural form of the compliant mechanism. The compliant mechanism applied to the micro-injector at present widely adopts a plane centralized compliant mechanism, and has the following problems:
(1) the fatigue problem is as follows: fatigue is a big cause of failure of the compliant mechanism, and is especially considered for compliant guide mechanisms that adopt high-frequency vibration strategies for puncturing. The stress concentration problems inherent in concentrated compliant mechanisms are more likely to cause fatigue failure of the mechanism.
(2) The problem of rigidity: the transverse vibration of the compliant mechanism depends on transverse rigidity in principle, and the plane mechanism can only increase the rigidity in the direction perpendicular to the plane by increasing the thickness, which can cause the coupling of the rigidity in each direction and can not enable the compliant mechanism to achieve an ideal rigidity ratio.
(3) Parasitic motion problems: the plane mechanism is asymmetric in circumferential direction, so that circumferential coupling errors caused by input cannot be eliminated, transverse parasitic motion is caused, and mechanical efficiency and transmission precision are reduced.
In addition, in the automatic puncturing process, due to the characteristics of high elasticity of cell membranes, geometric nonlinearity of the puncturing process and complex internal environment of cells, cell puncturing failure is caused or cell damage is large in the puncturing process, in order to improve the success rate of cell puncturing and the survival rate of the punctured cells, the cells need to be punctured in a shorter displacement and time, and the puncturing force and the puncturing speed are very high, so that force and position control is needed in cell puncturing. The existing force and position control applied to cell puncture needs to be switched between two different control laws, which may cause instability of the whole system, further cause long convergence time and cannot carry out force and position following control under the condition of high-frequency vibration; the disturbances such as hysteresis and friction of the piezoelectric actuator also present a great challenge to the existing force and position control, and these external disturbances greatly increase the steady-state force error and the steady-state position error. Therefore, the existing force and position control system has the defects of long convergence time, instability and large steady-state error, and cannot be well applied to cell puncture, particularly to high-frequency vibration puncture with more development prospects at present.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a cell microinjection device, which can reduce the transverse vibration of a needle head in the microinjection device and reduce the damage of cells in the process of cell puncture based on the guiding function of a hub type compliant mechanism. Meanwhile, the invention also provides a robust impedance control method, which is easier to control the cell puncture force and position and improves the stability of a control system.
The technical scheme is as follows: a cell microinjection device comprises a connecting rod, an encAN _ SNsulator connected with the connecting rod, a piezoelectric driver fixed in the encAN _ SNsulator, a microinjection needle, an injector and a hub type compliant mechanism, wherein two ends of the injector are respectively connected with the encAN _ SNsulator and the hub type compliant mechanism, and the microinjection needle is fixedly connected with the hub type compliant mechanism; the hub type compliant mechanism is provided with a reed type compliant unit for transmitting axial vibration to the micro-injection needle.
Has the advantages that: compared with the prior art, the invention has the advantages that: by adopting the distributed flexible mechanism, the rigidity ratio of the axial direction to the transverse direction can be increased, so that the transverse vibration caused by various errors such as assembly errors, processing errors and the like is reduced, the cell damage caused by the transverse vibration is fundamentally avoided, and the success rate of cell puncture is improved.
Furthermore, the hub type compliant mechanism is provided with two concentric hollow cylinders, the end surfaces of the two hollow cylinders are connected through a plurality of reed type compliant units, and the reed type compliant units are uniformly distributed along the circumferential direction of the two hollow cylinders. Compared with a planar compliant mechanism, parasitic motion is reduced.
Furthermore, the bending angle of the needle tip head of the micro-injection needle is 0 degree, and additional transverse vibration is prevented from being generated.
Furthermore, an L-shaped pore channel is arranged in the injector, one port of the pore channel is arranged on the side surface of the injector, and the microinjection needle penetrates through the compliant mechanism and is inserted into the other port of the pore channel. The liquid injection to the cells is realized through the arrangement of the pore channels.
Furthermore, a fixed chuck is arranged on the outer surface of the hub type compliant mechanism, the fixed chuck is movably connected with one end of the connecting rod in a limiting mode, and a pre-tightening force is applied to a piezoelectric driver fixed on the packaging device by adjusting the positions of the fixed chuck and the connecting rod.
Furthermore, a sealing ring is arranged at the interface of the pore channel and the micro-injection needle to prevent liquid leakage.
The invention also provides a technical scheme of the robust impedance control method of the cell microinjection device, which comprises the following steps:
a micro-force sensor is arranged at the bottom of the micro-injection needle, a displacement sensor is integrated in a piezoelectric driver, and the puncture force and the puncture position of the micro-injection needle are adjusted by a controller; the control system of the controller is provided with a control loop of the micro-injection needle, the control loop of the micro-injection needle is a position variable x, and when the needle tip of the micro-injection needle is contacted with an injected object, contact force f is generated, and f is contacted with ideal contact force fdDifference is made to obtain contact force error ef，efAs input to the impedance model, the position error e of the response is obtainedpFeeding back to a control loop of the micro-injection needle, and adjusting position input to ensure that the position of the micro-injection needle and the contact force between the micro-injection needle and the environment meet a preset ideal track; and a sliding mode function is designed in the impedance model to carry out self-adaptive control compensation so as to reduce the influence of external disturbance.
Further, the kinetic equation of the impedance model is:
wherein m isiIs a virtual effective mass, biIs the virtual effective damping coefficient, kiIs the virtual effective stiffness coefficient, kfIs the coefficient of external force, epIs the position error, efIs the error in the force or forces,
is the velocity error obtained by time-deriving the position error,
is the acceleration error obtained by time quadratic derivation of the position error.
Further, the steady-state position error and the steady-state contact error are analytically expressed as:
wherein k isiIs the virtual effective stiffness coefficient, kfIs an external force coefficient for adjusting the force error efThe weight of (a) is determined,
is the effective stiffness coefficient of the external environment, fdIs the ideal required force, xdIs the ideal desired displacement, xeIs the equilibrium position of the environment in the absence of interaction forces, epssIs the steady state position error, efssIs the steady state force error and n is the coefficient that varies according to the impedance model.
Furthermore, the slope k of the given displacement-time reference signal and the maximum value f of the given force-time signalmaxSatisfies the following conditions: k is more than or equal to 0.5 and less than or equal to 1, and f is more than or equal to 10mNmax≤40mN。
Has the advantages that: compared with the prior art, the robust impedance control method provided by the invention comprises the following steps: the problem of unstable system caused by control switching is avoided, the control on the cell puncture force and the cell puncture position is easier to complete by using the impedance module, the stability of the control system is improved, and the damage to the cells is reduced.
Drawings
FIG. 1 is a schematic perspective view of a microinjection apparatus according to the present invention;
FIG. 2 is a schematic perspective view of a connecting rod according to the present invention;
FIG. 3 is a schematic perspective view of an injector according to the present invention;
FIG. 4 is a schematic perspective view of a hub-type compliant mechanism according to the present invention;
FIG. 5 is a flow chart of the puncturing and injecting process of the cell microinjection apparatus according to the present invention;
FIG. 6 is a control framework of the robust impedance control method of the present invention;
FIG. 7 shows the ideal required force f in the present inventiond-time t diagram and ideal required force displacement xd-a time t-plot.
Detailed Description
The technical scheme of the invention is further explained by combining the attached drawings.
As shown in fig. 1 to 4, the cell microinjection apparatus of the present invention includes a connection rod 1, an encapsulator 2, a piezoelectric driver 3, an injector 4, a hub-type compliance mechanism 5, a fixed chuck 6, and a microinjection needle 7.
The connecting rod 1 comprises a threaded part 11, a connecting part 12 and a pressing part 13, wherein the threaded part 11 is connected with the fixed chuck 6, the fixed chuck 6 is fixed at the outer side of the hub type compliance mechanism 5, the threaded part 11 is also provided with a nut, and the relative position of the connecting rod 1 and the fixed chuck 6 is adjusted by rotating the nut; the pressing part 13 is abutted against one end of the packaging device 2, when the nut is screwed, a pre-tightening force is applied to the piezoelectric driver 3 in the packaging device 2, and the piezoelectric driver 3 is made of piezoelectric ceramics with a flexible operational amplifier mechanism.
The hub type compliant mechanism 5 is provided with two hollow concentric cylinders, namely an inner hollow cylinder threaded part 53 and an outer hollow cylinder 54, wherein the outer hollow cylinder 54 is fixedly connected to the fixed chuck 6 under the clamping force of the fixed chuck 6; the hub type compliance mechanism 5 further comprises a threaded portion 51 and a reed type compliance unit 52. The threaded part 51 is in threaded connection with the threaded part 41 to transmit high-frequency vibration of the piezoelectric driver 3, and the connection part of the threaded part 51 and the threaded part 41 is properly lubricated to reduce abrasion; the reed type compliant units 52 are respectively connected with the inner end surface of the outer hollow cylinder 54 and the outer end surface of the inner hollow cylinder 53, and are uniformly distributed along the circumferential direction, and the reed type compliant units 52 totally have 6 pieces and perform axial guide action on the movement direction transmitted to the hub type compliant mechanism 5; the micro-injection needle 7 penetrates through the inner hollow cylinder 53 and is attached to the inner side surface of the inner hollow cylinder 53, so that the micro-injection needle 7 is fixedly connected with the hub type compliant mechanism 5, and the micro-injection needle 7 vibrates along with the hub type compliant mechanism 5.
As shown in the flow chart of fig. 5, when cell puncture starts, the signal generator generates a high-frequency sinusoidal signal and obtains a filtered signal through the band-stop filter, the piezoelectric driver 3 fixed in the packaging device 2 starts to generate high-frequency vibration after receiving the signal, the vibration is transmitted to the injector 4 connected with the packaging device 2 under the action of the pre-tightening force of the connecting rod 1 in threaded connection with the fixed chuck 6, the injector 4 transmits the vibration to the hub type compliant mechanism 5 in threaded connection, and the axial vibration motion is transmitted to the micro-injection needle 7 under the guiding action of the reed type compliant unit 52, so that the needle tip of the micro-injection needle 7 generates high-frequency axial vibration and pierces cells to complete the cell puncture process. In the cell injection process, the liquid to be injected is injected into the pore channel of the injector 4 through an external injection device, the liquid is transferred to the hollow cavity inside the micro-injection needle 7 communicated with the pore channel 42 through the pore channel 42, and the liquid to be injected enters the cell which is subjected to cell puncture through the inner cavity of the micro-injection needle 7, so that the cell injection process is completed.
As shown in fig. 6, the robust impedance control method of the present invention is to use a controller to simultaneously adjust the penetration force and position of the micro-injection needle 7; a micro-force sensor is arranged at the bottom of the micro-injection needle 7, and a displacement sensor is integrated in the piezoelectric driver to obtain ideal required displacement xdAnd the ideal required piercing force fd(ii) a The control frame of the controller is provided with two closed-loop controls, wherein the inner loop is a position feedback loop, the position x of the microinjection needle is subjected to closed-loop control through the feedback controller, and the position x of the microinjection needle is a position variable; when the needle tip of the injection needle contacts with the injected object, a contact force f is generated, and the contact force f between the microinjection needle and the cell and the ideal demand force f are obtained through the force sensordObtaining delta x through an impedance model after difference is made, and taking the delta x as an input signalFeeding to a position feedback loop, and adjusting the position input to enable the position of the injection needle and the contact force of the injection needle and the environment to meet a preset ideal track.
Wherein, the difference between the contact force f and the ideal demand force f is obtained by an impedance model to obtain Δ x, which is a dynamic equation using model impedance, that is
(wherein, miIs a virtual effective mass, biIs the virtual effective damping coefficient, kiIs the virtual effective stiffness coefficient, kfIs the coefficient of external force, epIs the position error, efIs the error in the force or forces,
is the velocity error obtained by time-deriving the position error,
acceleration error obtained by performing time quadratic derivation on the position error), force error e in the puncture processfConverted into a position error epTherefore, the high-robustness puncture force and position control of the microinjection needle is achieved.
Specifically, the position error e in the actual process of cell puncturepSum force error efIs a relatively variable value, so that e in a steady state is generally usedpssAnd efssDescribing the steady state position error e in the process of the microscope injection needle puncturing the cell by the corresponding expressionpssSteady state force error efss. Wherein e ispssAnd efssThe expression is satisfied:
wherein k isiIs the virtual effective stiffness coefficient, kfIs an external force coefficient for adjusting the force error efThe weight of (a) is determined,
is the effective stiffness coefficient of the external environment, fdIs the ideal required force, xdIs the ideal desired displacement, xeIs the equilibrium position of the environment in the absence of interaction forces, epssIs the steady state position error, efssIs the steady state force error and n is the coefficient that varies according to the impedance model. In addition, a sliding mode function is designed in the impedance model to carry out adaptive control compensation so as to reduce the influence of external disturbance.
FIG. 7 is a graph showing the time-dependent changes of the input parameters of the control system, for example, in the case of a mouse oocyte puncture. The left graph is the ideal required force fdTime t diagram, right diagram ideal required force displacement xd-a time t-plot. For the left panel, the ideal force required to puncture mouse oocytes should be maintained at 20 mN; for the right plot, the slope k of the ideal desired displacement over time, i.e., the magnitude of the ideal puncture velocity, should be maintained at 0.7 m/s.
Claims (10)
1. A cell microinjection apparatus, comprising: the device comprises a connecting rod (1), an encAN _ SNsulator (2) connected with the connecting rod (1), a piezoelectric driver (3) fixed in the encAN _ SNsulator (2), and a micro-injection needle (7), and is characterized by further comprising an injector (4) and a hub type compliant mechanism (5), wherein two ends of the injector (4) are respectively connected with the encAN _ SNsulator (2) and the hub type compliant mechanism (5), and the micro-injection needle (7) is fixedly connected with the hub type compliant mechanism (5); the hub type compliant mechanism (5) is provided with a reed type compliant unit (52) for transmitting axial vibration to the micro-injection needle (7).
2. The cell microinjection apparatus according to claim 1, wherein the hub-type compliance mechanism (5) has two concentric hollow cylinders, the end surfaces of the two hollow cylinders are connected by a plurality of reed-type compliance units (52), and the plurality of reed-type compliance units (52) are uniformly distributed along the circumferential direction of the two hollow cylinders.
3. Cell microinjection apparatus according to claim 1, wherein the tip head of the microinjection needle (7) is bent at an angle of 0 °.
4. The cell microinjection apparatus according to claim 1, wherein the injector (4) is provided therein with an "L" shaped channel (42), one end of the channel (42) is disposed at a side of the injector (4), and the microinjection needle (7) penetrates the compliant mechanism (5) and is inserted into the other end of the channel (42).
5. The cell microinjection apparatus according to claim 1, further comprising a fixed collet (6) located on an outer surface of the hub-type compliance mechanism (5), wherein the fixed collet (6) is movably connected to one end of the connecting rod (1) in a limited manner.
6. Cell microinjection apparatus according to claim 4, wherein a sealing ring is installed at the interface of the duct (42) and the microinjection needle (7).
7. A robust impedance control method for cell microinjection apparatus according to any of claims 1 to 6, characterized in that a micro force sensor is installed at the bottom of the microinjection needle (7), a displacement sensor is integrated inside the piezoelectric actuator (3), and a controller is used to adjust the puncture force and position of the microinjection needle (7) at the same time; the control system of the controller is provided with a control loop of the micro-injection needle, the control loop of the micro-injection needle is a position variable x, and when the needle tip of the micro-injection needle is contacted with an injected object, contact force f is generated, and f is contacted with ideal contact force fdDifference is made to obtain contact force error ef，efAs input to the impedance model, the position error e of the response is obtainedpFeeding back to a control loop of the micro-injection needle, and adjusting position input to ensure that the position of the micro-injection needle and the contact force between the micro-injection needle and the environment meet a preset ideal track; and a sliding mode function is designed in the impedance model to carry out self-adaptive control compensation so as to reduce the influence of external disturbance.
8. The robust impedance control method of claim 7, wherein the dynamical equation of the impedance model is:
wherein m isiIs a virtual effective mass, biIs the virtual effective damping coefficient, kiIs the virtual effective stiffness coefficient, kfIs the coefficient of external force, epIs the position error, efIs the error in the force or forces,
is the velocity error obtained by time-deriving the position error,
is the acceleration error obtained by time quadratic derivation of the position error.
9. The robust impedance control method of claim 8, wherein the analytical expression of the steady state position error and the steady state contact error is:
wherein k isiIs the virtual effective stiffness coefficient, kfIs an external force coefficient for adjusting the force error efThe weight of (a) is determined,
is the effective stiffness coefficient of the external environment, fdIs the ideal required force, xdIs the ideal desired displacement, xeIs the equilibrium position of the environment in the absence of interaction forces, epssIs the steady state position error, efssIs the steady state force error and n is the coefficient that varies according to the impedance model.
10. The robust impedance control method of a cell microinjection apparatus according to claim 7, wherein the given displacement-time reference signal slope k and the given maximum value f of the force-time signalmaxSatisfies the following conditions: k is more than or equal to 0.5 and less than or equal to 1, and f is more than or equal to 10mNmax≤40mN。
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| CN114107023A (en) * | 2021-11-23 | 2022-03-01 | 南京航空航天大学 | Piezoelectric driving cell microinjection device and self-adaptive compliance control method thereof |
| CN114369516A (en) * | 2021-12-27 | 2022-04-19 | 华南理工大学 | Membrane puncturing device based on piezoelectric superstructure strong-modal damping compliant guide mechanism |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114107023A (en) * | 2021-11-23 | 2022-03-01 | 南京航空航天大学 | Piezoelectric driving cell microinjection device and self-adaptive compliance control method thereof |
| CN114107023B (en) * | 2021-11-23 | 2024-03-19 | 南京航空航天大学 | Piezoelectric driving cell microinjection device and self-adaptive compliant control method thereof |
| CN114369516A (en) * | 2021-12-27 | 2022-04-19 | 华南理工大学 | Membrane puncturing device based on piezoelectric superstructure strong-modal damping compliant guide mechanism |
| CN114369516B (en) * | 2021-12-27 | 2023-12-22 | 华南理工大学 | Film puncturing device based on piezoelectric superstructure strong modal damping compliant guide mechanism |
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Application publication date: 20211102 |