阅读说明：本技术 一种精确控制药物分子的装置及方法 (Device and method for accurately controlling drug molecules ) 是由 袁志山 吴丹丹 王成勇 于 2019-09-04 设计创作，主要内容包括：本发明提供一种精确控制药物分子的装置,所述装置包括上粗下细的筒状玻璃针、纳米孔芯片及控制驱动电路；控制驱动电路包括用于统计药物分子数量的计数测量电路、用于驱动药物分子穿过纳米孔芯片的第一驱动电路及用于驱动药物分子注入待检测细胞的第二驱动电路；纳米孔芯片水平内嵌于玻璃针的内侧壁上,玻璃针的内腔填充有电解质溶液。本发明还提供一种精确控制药物分子的方法,操作方法简单、工作效率高且又能从药物分子级别实现药物用量的精确控制。(The invention provides a device for accurately controlling drug molecules, which comprises a cylindrical glass needle with a thick upper part and a thin lower part, a nanopore chip and a control drive circuit; the control driving circuit comprises a counting measurement circuit for counting the number of the drug molecules, a first driving circuit for driving the drug molecules to pass through the nanopore chip and a second driving circuit for driving the drug molecules to be injected into the cell to be detected; the nanopore chip is horizontally embedded in the inner side wall of the glass needle, and the inner cavity of the glass needle is filled with electrolyte solution. The invention also provides a method for accurately controlling the drug molecules, which has simple operation method and high working efficiency and can realize accurate control of the drug dosage from the level of the drug molecules.)

1. The device for accurately controlling the drug molecules is characterized by comprising a cylindrical glass needle (1), a nanopore chip (2) and a control drive circuit (3); the control drive circuit (3) comprises a counting measurement circuit (31) for counting the number of the drug molecules (5), a first drive circuit (32) for driving the drug molecules (5) to pass through the nanopore chip (2), and a second drive circuit (33) for driving the drug molecules (5) to be injected into the cell (6) to be detected; the nanopore chip (2) is horizontally fixed on the inner side wall of the glass needle (1), an electrolyte solution (4) is filled in the inner cavity of the glass needle (1), and drug molecules (5) are dissolved in the electrolyte solution (4).

2. The device for precisely controlling drug molecules according to claim 1, wherein the glass needle (1) is thick at the top and thin at the bottom and comprises an upper end barrel (11) and a lower end barrel (12), the upper end barrel (11) is hollow cylindrical and has a diameter of 2-5 mm; the lower end cylinder (12) is in a hollow circular truncated cone shape with a thick upper part and a thin lower part, and the caliber of the tip of the lower end cylinder (12) is 1-10 mu m.

3. The device for precisely controlling drug molecules according to claim 2, wherein the nanopore chip (2) is any one of a silicon chip, a silicon nitride chip, and a silicon dioxide chip; a nanopore (21) is drilled at the center of the nanopore chip (2), and the diameter of the nanopore (21) is 10-200 nm.

4. The device for precisely controlling drug molecules according to claim 3, wherein the glass needle (1) is provided with a micropump, and the micropump comprises a first driving micropump (321) for driving the drug molecules (5) to pass through the nanopore (21) and a second driving micropump (333) for driving the drug molecules (5) to be injected into the cell (6) to be detected; the first driving micropump (321) is arranged at the top end of the glass needle (1); the second driving micropump (333) is arranged on the lower end cylinder (12), and the distance between the second driving micropump and the tip of the lower end cylinder (12) is no more than 1/3 of the vertical height of the lower end cylinder (12).

5. The device for precise control of drug molecules according to claim 4, wherein the counting and measuring circuit (31) comprises a first power supply (311), a current measuring device (312), and a first electrode (313) and a second electrode (314), wherein the first electrode (313) and the second electrode (314) are located on the same axis with the center of the nanopore (21); the first electrode (313) arranged above the nanopore chip (2) is electrically connected with the positive potential end of the first power supply (311) through the current measuring device (312), and the second electrode (314) arranged below the nanopore chip (2) is electrically connected with the negative potential end of the first power supply (311).

6. The device for precisely controlling drug molecules according to claim 5, wherein the first driving circuit (32) comprises a first switch (322) and a micro valve (323) disposed in the inner cavity of the glass needle (1), the first driving micro pump (321) is electrically connected to the positive potential terminal of the first power source (311) through the first switch (322), and the negative potential terminal of the first power source (311) is electrically connected to one terminal of the micro valve (323).

7. The apparatus for precisely controlling drug molecules according to claim 6, wherein the second driving circuit (33) comprises a second power source (331) and a second switch (332), one end of the micro valve (323) is further connected to a positive potential terminal of the second power source (331) through the second switch (332), and a negative potential terminal of the second power source (331) is electrically connected to one end of the second driving micro pump (333).

8. The device for precisely controlling drug molecules according to claim 5, wherein the current measuring device (312) is a picoampere meter, and when any one drug molecule (5) passes through the nanopore (21) of the nanopore chip (2), the reading of the current measuring device (312) is reduced, so that the current for reducing the reading of the current measuring device (312) is a blocking current.

9. The device for precisely controlling drug molecules in accordance with claim 7, wherein the voltages of the first power source (311) and the second power source (331) are 100-300 mV.

10. A method for precisely controlling a drug molecule, which is performed by the device for precisely controlling a drug molecule according to any one of claims 7 to 9, comprising the steps of:

s1, injecting an electrolyte solution (4) containing drug molecules (5) into an inner cavity of a glass needle (1) by using an injection needle;

s2, closing a first switch (322) of a first driving circuit (32), driving the electrolyte solution (4) by a first driving micropump (321) and a micro valve (323) together, and driving the drug molecules (5) to downwards penetrate through the nano holes (21);

s3, observing the reading of the current measuring device (312), and recording the number of times of reading reduction of the current measuring device (312) as the number of the drug molecules (5) passing through the nanopore (21);

s4, when the number of the drug molecules (5) passing through the nano holes (21) reaches the number of the needed drug molecules (5), disconnecting the first switch (322) of the first driving circuit (32), and stopping the downward movement of the drug molecules (5) which are positioned above the nano holes (21) and do not pass through the nano holes (21);

s5, a second switch (332) of a second driving circuit (33) is closed, a second driving micropump (333) and the micro valve (4) jointly drive the electrolyte solution (323) and drive the drug molecules (5) which penetrate through the nano holes (21) to move downwards, and the tip of the glass needle (1) is used for conveying the drug molecules (5) into the cell (6) to be detected.

Technical Field

The invention relates to the technical field of medical appliance manufacturing, in particular to a device and a method for accurately controlling drug molecules.

Background

In recent years, with the importance of bioengineering, cell engineering and genetic engineering techniques being increased, the importance of the bioengineering, cell engineering and genetic engineering techniques is increasing in the development of national economy.

In the engineering technologies, such as cell operation technologies of cell nucleus transplantation, intracellular drug injection, embryo injection, cell hybridization and the like, drug injection needs to be carried out on cells, the basic injection tool is a glass microneedle, namely, the drugs are injected into the glass microneedle, then, the needle point is penetrated into the cells through an accurate positioning and displacement adjusting system to realize micro-injection of the drugs, and the optimal dose is determined according to the accurate comparison of physical therapy effects of different drug doses applied in the process. The length-diameter ratio of the glass microneedle for injection is large, large resistance needs to be overcome during injection, and if the glass microneedle is manually operated in the traditional mode, the problem that the operation pressure is difficult to control occurs. Therefore, in the modern fields of cytology and molecular biology, how to efficiently and conveniently carry out quantitative drug injection on cells becomes a current research hotspot.

At present, the cell medicine quantitative control injection method mainly comprises two methods:

1. quantitative injection of the drug is achieved in the cell injector using pneumatic means. In the process, manual adjustment and microscopic observation of workers are required to be carried out simultaneously, and the manual adjustment and microscopic observation are mainly realized by the technology of the operators, so that the method has the problems of poor accuracy, low efficiency and high cost in controlling the dosage of the drug molecules.

2. The drug quantitative injection is realized by means of a micro-fluid system. The method has higher precision of controlling the dosage of the medicine, but has higher manufacturing cost and is difficult to meet the requirement of industrialization.

Therefore, it is very important to research a device which has simple operation method and high working efficiency and can realize accurate control of the drug dosage from the drug molecular level.

Disclosure of Invention

In order to overcome the defects that the prior art is inconvenient to operate, has poor control dosage precision on medicines, and has high production cost in the technology capable of realizing accurate control of the medicine dosage, the invention provides a device and a method for accurately controlling medicine molecules, and the purposes of convenient operation, low production cost and accurate control of the dosage of the medicine molecules are achieved.

In order to achieve the technical effects, the technical scheme of the invention is as follows:

a device for accurately controlling drug molecules comprises a cylindrical glass needle 1, a nanopore chip 2 and a control drive circuit 3; the control drive circuit 3 comprises a counting measurement circuit 31 for counting the number of the drug molecules 5, a first drive circuit 32 for driving the drug molecules 5 to pass through the nanopore chip 2, and a second drive circuit 33 for driving the drug molecules 5 to be injected into the cell 6 to be detected; the nanopore chip 2 is horizontally fixed on the inner side wall of the glass needle 1 by adopting the existing anodic bonding method: the inner side wall of the glass needle 1 is contacted with the edge of the nanopore chip 2 made of metal material, and after direct current is introduced during heating, the edge of the nanopore chip 2 is firmly connected to the inner side wall of the glass needle 1; the inner cavity of the glass needle 1 is filled with electrolyte solution 4, and drug molecules 5 are dissolved in the electrolyte solution 4.

Preferably, the glass needle 1 is thick at the top and thin at the bottom and comprises an upper end cylinder 11 and a lower end cylinder 12, wherein the upper end cylinder 11 is in a hollow cylindrical shape and has a diameter of 2-5 mm; the lower end barrel 12 is in a hollow circular truncated cone shape with a thick upper part and a thin lower part, and the caliber of the tip of the lower end barrel 12 is 1-10 mu m.

Preferably, the nanopore chip 2 is any one of a silicon chip, a silicon nitride chip and a silicon dioxide chip; the technology for punching the nanopore 21 at the center of the nanopore chip 2 can be any one of a focused ion beam, a high-energy electron beam and a laser, and the diameter of the nanopore 21 is 10-200 nm.

Preferably, the glass needle 1 is provided with a micropump, and the micropump comprises a first driving micropump 321 for driving the drug molecules 5 to pass through the nanopore 21 and a second driving micropump 333 for driving the drug molecules 5 to be injected into the cell 6 to be detected; the first driving micropump 321 is arranged at the top end of the glass needle 1; the second driven micropump 333 is disposed on the lower barrel 12 at a distance from the tip of the lower barrel 12 that does not exceed the vertical height 1/3 of the lower barrel 12.

Preferably, the counting and measuring circuit 31 comprises a first power source 311, a current measuring device 312, and a first electrode 313 and a second electrode 314, wherein the first electrode 313 and the second electrode 314 are located on the same axis with the center of the nanopore 21, so that the current measuring device 312 can accurately measure the measuring current I when the drug molecule 5 passes through the nanopore 21; a first electrode 313 disposed above the nanopore chip 2 is electrically connected to a positive potential terminal of the first power source 311 through a current measuring device 312, and a second electrode 314 disposed below the nanopore chip 2 is electrically connected to a negative potential terminal of the first power source 311, the current measuring device312 is a picoampere meter, when any drug molecule 5 passes through the nanopore 21 of the nanopore chip 2, the reading of the current measuring device 312 is decreased, so that the current of the decreased reading of the current measuring device 312 is a blocking current I_downAfter the inner cavity of the glass needle 1 is filled with the electrolyte solution 4, the counting and measuring circuit 31 becomes a closed and conductive loop, and when the drug molecule 5 does not pass through the nanopore 21, the reading of the current measuring device 312 is the unchanged initial state current I₀When any one drug molecule passes through the nanopore 21, the measurement current I is changed from the original initial state current I₀Down to a blocking current I_downWhen the drug molecule 5 passes through the nanopore 21, the measurement current I will be determined by the occlusion current I_downIs restored to the initial state current I₀Therefore, by observing the current reading of the current measuring device 312 and recording the number of drops of the measured current I, the worker can accurately count the number of drug molecules 5 passing through the nanopore 21, thereby further accurately controlling the amount of drug molecules.

Preferably, the first driving circuit 32 includes a first switch 322 and a micro valve 323 disposed in the inner cavity of the glass needle 1, the first driving micro pump 321 is electrically connected to the positive potential end of the first power source 311 through the first switch 322, the negative potential end of the first power source 311 is electrically connected to one end of the micro valve 323, when the first switch 322 is closed, the first driving circuit 32 is turned on, the first driving micro pump 321 and the micro valve 323 disposed in the inner cavity of the glass needle 1 are started, because the first driving micro pump 321 is disposed at the upper end of the glass needle 1, the first driving micro pump 321 is electrically connected to the positive potential end of the first power source 311 through the first switch 322, the first driving micro pump 321 generates force, and ions in the electrolyte solution 4 drive the drug molecules 5 to move downward, reach the upper side of the nanopore 21, and further sequentially pass through the nanopore 21; when the first switch 322 is turned off, the first driving circuit 32 cannot form a closed loop, the ions in the electrolyte solution 4 cannot move, and the drug molecules 5 also stop moving downward.

Preferably, the second driving circuit 33 comprises a second power source 331 and a second switch 332, one end of the micro valve 323 is further connected to the positive potential end of the second power source 331 through the second switch 332, and the negative potential end of the second power source 331 is electrically connected to one end of the second driving micro pump 333; when the second switch 332 is closed, the second driving circuit 33 is turned on, the second driving micropump 333 and the microvalve 323 are started, and since the position of the second driving micropump 321 in the second driving circuit 33 relative to the microvalve 323 is opposite to the position of the first driving micropump 321 relative to the microvalve 323, and the second driving micropump 333 is disposed at the lower end of the glass needle 1, the second driving micropump 333 is electrically connected to the negative potential end of the second power source 331 through the second switch 332, the second driving micropump 333 generates force, ions in the electrolyte solution 4 drive the drug molecules 5 which have passed through the nanopore 21 to continue to move downwards, and the drug molecules 5 are transported into the cell 6 to be detected by using the tip of the glass needle 1.

Preferably, the voltages of the first power source 311 and the second power source 331 are both 100 mV to 300 mV.

The invention also provides a method for accurately controlling the drug molecules, which is realized by the device for accurately controlling the drug molecules and comprises the following steps:

s1, injecting an electrolyte solution 4 containing drug molecules 5 into an inner cavity of a glass needle 1 by using an injection needle;

s2, closing a first switch 322 of the first driving circuit 32, opening a first driving micro pump 321 and a micro valve 323, driving the electrolyte solution 4 by the first driving micro pump 321 and the micro valve 323 together, and driving the drug molecules 5 to downwards pass through the nano holes 21;

s3, observing the reading of the current measuring device 312, and recording the number of times of reading reduction of the current measuring device 312 as the number of the drug molecules 5 passing through the nanopore 21;

s4, when the number of the drug molecules 5 passing through the nano-holes 21 reaches the number of the needed drug molecules 5, disconnecting the first switch 322 of the first driving circuit 32, stopping the first driving micropump 321 and the microvalve 323, and stopping the downward movement of the drug molecules 5 which are positioned above the nano-holes 21 and do not pass through the nano-holes 21;

s5, closing a second switch 332 of the second driving circuit 33, opening a second driving micropump 333 and a micro valve 323, driving the electrolyte solution 4 by the second driving micropump 333 and the micro valve 323 together, driving the drug molecules 5 passing through the nanopores 21 to move downwards, and conveying the drug molecules 5 into the cells 6 to be detected by using the tip of the glass needle 1.

Compared with the prior art, the technical scheme of the invention has the beneficial effects that:

(1) the device provided by the invention adopts a counting and measuring circuit, records the occurrence frequency of the blocking current, namely the number of the drug molecules passing through the nanopore, by comparing the initial state current with the blocking current from the angle of observing the current change, and realizes the accurate control of the quantitative drug injection of the cells on the molecular level, thereby determining the drug dosage with the optimal physical therapy effect.

(2) The device provided by the invention can realize accurate control of the number of the drug molecule via holes only by controlling the driving circuit, does not need to use other auxiliary devices with high manufacturing cost, such as a microscope and the like, is simple to operate, has no strict requirements on the operation technology of workers, and can effectively improve the production efficiency.

Drawings

Fig. 1 is a schematic diagram of the overall structure of the device for precisely controlling drug molecules according to the present invention.

Fig. 2 is a schematic structural diagram of a glass needle of the device for precisely controlling drug molecules according to the present invention.

Fig. 3 is a schematic structural diagram of a nanopore chip according to the present invention.

FIG. 4 is a flow chart of the method for precisely controlling drug molecules according to the present invention.

Fig. 5 is a diagram of the measured current during the use of the device according to the invention.

Detailed Description

The drawings are for illustrative purposes only and are not to be construed as limiting the patent;

for the purpose of better illustrating the embodiments, certain features of the drawings may be omitted, enlarged or reduced, and do not represent the size of an actual product;

it will be understood by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted.

The technical solution of the present invention is further described below with reference to the accompanying drawings and examples.