CN110108627A - A kind of cell in-situ observation and operating device based on microsphere lens - Google Patents

Abstract

The invention discloses a kind of cell in-situ observation and operating device based on microsphere lens.The device includes microscope module, article carrying platform, the first microoperation manipulator, the second microoperation manipulator and microsphere lens.Cell in-situ observation and operating device provided by the embodiment of the present invention based on microsphere lens is using microsphere lens as the tool of cellular micromanipulation, it realizes and picks up almost not damaged to cell in cell processes and do not change the effect of cellular morphology, improve the success rate of cellular micromanipulation.

Description

A device for in situ observation and manipulation of cells based on microsphere lenses

technical field

The invention relates to the technical field of biomedical related surgical devices, in particular to a cell in situ observation and operation device based on a microsphere lens.

Background technique

With the advancement of science and technology, scientific research and clinical applications in the fields of biology and medicine have gradually developed towards microcosm and precision, among which cell micromanipulation is the technical guarantee to promote this development trend. Artificial fertilization, cloning, genetic engineering and drug discovery all rely on the precise manipulation of biological cells. The most effective and easiest way to make transgenic animals is to use microinjection technology, that is, to inject exogenous genes into the active fertilized eggs of animals with a micro-operating system, so as to cultivate new varieties. This micromanipulation process is an important link in the research and cultivation of transgenic animals in the biological field, and it is also a link that requires high operating precision. In this microscopic environment, manipulation of these cells requires submillimeter tools and must be done under a microscope, which is currently done manually.

Another large class of micromanipulation processes in biological and genetic engineering is microdissection. The basic structure and driving principle of the device for completing the cutting operation are the same as those for the micro-injection operation, and the micro-cutting device only needs a micro-cutting knife. The tip portion of the microcutting knife is also submicron in size. The displacement and positioning accuracy of the micro-manipulation device are higher than those of the micro-injection device. Micro-cutting operation has very important applications in biomedical research, especially in high-tech fields such as botany research, genetic engineering research, and establishment of DNA libraries of animals and plants (including humans). Like microinjection, microdissection is currently performed manually. Since the microdissection operation requires higher precision than the microinjection operation, it is more difficult to perform manually.

The technical level of cell micromanipulation directly determines the sophistication and success of relevant scientific research and clinical applications. The key to cell micromanipulation is the efficient pick-up, release, and movement of cells, as well as the firm fixation of cells during manipulation. At present, the most widely used cell manipulation tool is the micropipette. The earliest Yasuyuki Kimura et al. used the micropipette to achieve egg cell single sperm injection. The micropipette picks up, transfers and immobilizes cells by negative pressure adsorption. The cell puncture and injection operations rely on the local adsorption of the micropipette port to the cells to fix the cells, and cooperate with the puncture needle or injection needle to complete the operation task. The control of negative pressure depends entirely on the experience and proficiency of the operator. The operation process requires long-term concentration, and it is difficult to achieve a successful one-time picking and release of cells. In addition, during the operation, it is easy to cause excessive local negative pressure and damage cells, causing the operation to fail. Furthermore, in the process of operation, the adsorption effect of local negative pressure cannot firmly fix the cells, let alone limit the deformation of the cells, the operation precision is low, and it is impossible to realize finer cell operations inside the cells. In addition, the single operation mode of the micropipette cannot meet more complex, flexible and diverse cell operations. In addition to the objective factors of easy deformation and damage of the manipulated cells themselves, these operational difficulties are ultimately due to the fact that the operation mode, flexibility and motion control of cell micromanipulation tools cannot meet the requirements of cell micromanipulation.

Therefore, in view of the above technical problems, it is necessary to provide a cell in situ observation and manipulation device based on microsphere lenses that can avoid the low flexibility, low operation precision and high failure rate of the negative pressure adsorption method.

Contents of the invention

In view of this, the purpose of the embodiments of the present invention is to provide a cell in situ observation and manipulation device based on a microsphere lens that can avoid the low flexibility, low operation precision and high failure rate of the negative pressure adsorption method. The cell in-situ observation and manipulation device based on the microsphere lens provided by the embodiment of the present invention uses the microsphere lens as a tool for cell micromanipulation to achieve the effect of almost no damage to the cell and no change in the cell shape during the process of picking up the cell, which improves the Success rate of cellular micromanipulation.

In order to achieve the above object, the technical solution provided by an embodiment of the present invention is as follows: a microsphere lens-based cell in situ observation and operation device, including: a microscope module, used to magnify the cell to a preset multiple; an object loading platform, Located under the eyepiece of the microscope module, it is used to place the vessel; the first micro-manipulation manipulator is located on the first side of the microscope module, and is used to perform the first operation on the cells in the vessel; the second micro-manipulation manipulator , located on the second side of the microscope module, the first side and the second side are symmetrical about the central axis of the microscope module; the microsphere lens is arranged on the probe of the second micro-manipulator A needle end for performing a second manipulation of the cells within the vessel.

As a further improvement of the present invention, the object loading platform has degrees of freedom in two directions.

As a further improvement of the present invention, the object loading platform can move along the X-axis direction and along the Y-axis direction respectively.

As a further improvement of the present invention, the structure of the first micro-manipulator is the same as that of the second micro-manipulator.

As a further improvement of the present invention, the first micro-manipulator includes: a probe, a component for cell manipulation; a probe extension rod, one end of the probe extension rod is fixedly connected to the probe, and to extend the length of the probe; the angle adjustment mechanism is fixedly connected to the other end of the probe extension rod for adjusting the angle of the probe; the main body is fixedly connected to the angle adjustment mechanism; the first mobile platform, connected with the main body, used to adjust the position of the probe in the first direction; the second mobile platform, connected with the first mobile platform, used to adjust the position of the probe in the second direction; the third mobile platform , connected with the second mobile platform, for adjusting the position of the probe in the third direction.

As a further improvement of the present invention, the first direction, the second direction and the third direction are respectively perpendicular to each other.

As a further improvement of the present invention, the microsphere lens is fixed on the probe end of the second micro-manipulator by glue.

As a further improvement of the present invention, the first operation includes a cell puncture operation, a cell removal operation and/or a cell removal operation.

As a further improvement of the present invention, the second operation includes bonding and picking up cells.

As a further improvement of the present invention, the microsphere lens is a barium titanate glass lens.

The present invention has the following advantages:

The in-situ cell observation and manipulation device based on the microsphere lens provided by the embodiment of the present invention uses the microsphere lens as a tool for cell micromanipulation, and realizes picking up cells with almost no damage and no shape change, and conveniently realizes sample sampling , and get rid of the shackles of the substrate in the prior art. Further, the cell in situ observation and manipulation device based on the microsphere lens utilizes the microsphere lens as a tool for cell micromanipulation, providing sufficient space for manipulation of the observed cell. Furthermore, the in situ cell observation and manipulation device based on the microsphere lens uses the microsphere lens as a tool for cell micromanipulation. The microsphere lens can realize super-resolution imaging of the imaging sample under the microscope, and can observe the cells more finely. Furthermore, the cell in situ observation and manipulation device based on the microsphere lens uses the microsphere lens as a tool for cell micromanipulation. Through the microsphere lens, the shape of the cell can be observed in real time, and then the first micromanipulator can be used to in situ Realize convenient operations such as screening, puncturing, and elimination of cells.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only These are some embodiments described in the present invention. Those skilled in the art can also obtain other drawings based on these drawings without creative work.

Fig. 1 is a partial structural schematic diagram of a cell in situ observation and operation device based on a microsphere lens provided by an embodiment of the present invention;

Fig. 2 is a schematic structural view of the loading platform in the embodiment shown in Fig. 1;

Fig. 3 is the structural representation of the first micro-manipulator in the embodiment shown in Fig. 1;

Fig. 4 is a schematic structural view of the angle adjustment mechanism in the embodiment shown in Fig. 3;

Fig. 5 is the structural representation of microsphere lens and probe in the embodiment shown in Fig. 1;

Fig. 6 (a) is the state diagram of the second operation;

Fig. 6 (b) is the enlarged schematic view of the part shown in b in Fig. 6 (a);

FIG. 7( a ) to FIG. 7( c ) are schematic diagrams of states of different first operations.

Explanation of the marks in the attached drawings:

100. Cell in situ observation and manipulation device based on microsphere lens 10. Microscope module

21. The first micro-manipulator 22. The second micro-manipulator 30. Vessels

40. Object loading platform 50. Microsphere lens 42. Platform

43. Y-axis micrometer 45. X-axis micrometer 210. Main rod

211, the first mobile platform 213, the second mobile platform 215, the third mobile platform

212, angle adjustment mechanism 2121, first connecting part 2123, second connecting part

2124, angle adjustment knob 214, probe extension rod 216, first probe

60. Glue 213. The second mobile platform 215. The third mobile platform

226. Second Probe

Detailed ways

In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described The embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts shall fall within the protection scope of the present invention.

Referring to FIG. 1 , the first embodiment of the present invention provides a device 100 for in situ observation and manipulation of cells based on a microsphere lens. The device 100 includes a microscope module 10 , an object loading platform 40 , a first micro-manipulation manipulator 21 , a second micro-manipulation manipulator 22 and a microsphere lens 50 .

The microscope module 10 is used to magnify the cells to a preset multiple. The microscope module 10 can adopt a general optical microscope. Optical microscope specifically includes eyepiece, objective lens, coarse quasi-focus helix, fine quasi-focus helix, clamp, aperture, shutter, converter, mirror, mirror arm, lens barrel, mirror base, condenser, diaphragm (In Fig. 1, each part is not labeled separately, and not all parts are fully shown).

Continuing to refer to FIG. 1 , the object loading platform 40 is located below the eyepiece of the microscope module 10 . The loading platform 40 is used for placing the vessel 30 . In this embodiment, the vessel 30 contains an aqueous solution, and cells dispersed in the aqueous solution. Referring to FIG. 2 , the loading platform 40 has degrees of freedom in two directions, specifically, the loading platform 40 can move along the X-axis direction and along the Y-axis direction respectively. The loading platform 40 includes a platform 42 , an X-axis micrometer 45 located on one side of the platform 42 , and a Y-axis micrometer 43 located on the other side adjacent to the platform 42 . By rotating the X-axis micrometer 45, the position of the platform 42 in the X-axis can be precisely moved, thereby driving the position of the vessel 30 on the platform 42, and facilitating the picking operation of the cells by the microsphere lens 50; by rotating the Y-axis micrometer 43, it can be precisely The position of the platform 42 in the Y-axis is moved accordingly, thereby driving the position of the vessel 30 on the platform 42 to facilitate the picking operation of the cells by the microsphere lens 50 . In this embodiment, the X axis is in the direction of the X double-headed arrow marked in FIG. 2 , and the Y axis is in the direction of the Y double-headed arrow marked in FIG. 2 .

Continuing to refer to FIG. 1 , the first micro-manipulation manipulator 21 is located on the first side of the microscope module 10 for performing a first operation on the cells in the vessel 30 . The second micro-manipulator 22 is located on the second side of the microscope module 10 , wherein the first side and the second side are symmetrical about the central axis of the microscope module 10 . The first micro-manipulation manipulator 21 and the second micro-manipulation manipulator 22 are respectively located on two opposite sides of the microscope module 10 , and they are opposite to each other. The object loading platform 40 is located between the first micro-manipulation manipulator 21 and the second micro-manipulation manipulator 22 .

In this embodiment, the structure of the first micro-manipulator 21 is the same as that of the second micro-manipulator 22 . Therefore, only the first micro-manipulation manipulator 21 is taken as an example to introduce the structure in detail. For convenience, the structure numbers in the first micro-manipulation manipulator 21 and the structure numbers in the second micro-manipulation manipulator 22 also correspond. Specifically, for example: the number of the probe of the first micro-manipulation manipulator 21 is the first probe 216 , and the number of the probe of the second micro-manipulation manipulator 22 is the second probe 226 .

Referring to Fig. 3, the first micro-manipulator 21 includes a first probe 216 for cell manipulation, a probe extension rod 214, an angle adjustment mechanism 212, a main body 210, a first mobile platform 211, and a second mobile platform 213 , the third mobile platform 215 . One end of the probe extension rod 214 is fixedly connected to the first probe 216 , and the other end of the probe extension rod 214 is fixedly connected to the angle adjustment mechanism 212 . The probe extension rod 214 is used to extend the length of the first probe 216 in a preset direction. The angle adjustment mechanism 212 is used to adjust the angle of the first probe. The main body 210 is fixedly connected with the angle adjustment mechanism 212 .

The first mobile platform 211 is connected with the main body 210, and the first mobile platform 211 can move along a first direction. The first moving platform 211 moves along the first direction, drives the main body 210 to move along the first direction, thereby drives the angle adjustment mechanism 21 , the probe extension rod 214 and the first probe 216 to move along the first direction. The first moving platform 211 is used to adjust the position of the first probe 216 in the first direction. The second mobile platform 213 is connected with the first mobile platform 211, and the second mobile platform 213 can move along the second direction. The second moving platform 213 moves along the second direction, which can drive the first moving platform 211 to move along the second direction, and then drive the first probe 216 to move along the second direction. The second moving platform 213 is used to adjust the position of the first probe 216 in the second direction. The third mobile platform 215 is connected to the second mobile platform 213, and the third mobile platform 215 can move along a third direction. The third mobile platform 215 moves along the third direction, which can drive the second mobile platform 213 to move along the third direction, and further drives the first mobile platform 211 to move along the third direction, thereby driving the first probe 216 to move along the third direction. The third moving platform 215 is used to adjust the position of the first probe 216 in the third direction. In this embodiment, the first direction, the second direction and the third direction are respectively perpendicular to each other. Specifically, if the Cartesian coordinate system is established on the paper shown in FIG. 3 , the first direction is the Z direction, the second direction is the Y direction, and the third direction is the X direction. The first mobile platform 211 , the second mobile platform 213 and the third mobile platform 215 constitute a three-dimensional manipulator, and by moving the three-dimensional manipulator, the probe can reach any position in space. In other embodiments, the first mobile platform 211 may also be fixedly connected to the angle adjustment mechanism 212 directly, that is, the main body 210 is a part of the first mobile platform 211 or the main body 210 is omitted.

As shown in FIG. 4 , the first probe 216 and the probe extension rod 214 are installed at the end of the angle adjustment mechanism 212 . The angle adjustment mechanism 212 includes a first connection part 2121 , a second connection part 2123 and an angle adjustment knob 2124 . One end of the first connecting part 2121 is fixedly connected to the main body 210 , and the other end is movably connected to one end of the second connecting part 2123 through the angle adjustment knob 2124 . One end of the second connecting part 2123 is movably connected to the other end of the first connecting part 2121 through the angle adjustment knob 2124 , and the other end is fixedly connected to the probe extension rod 214 . The first probe 216 is tilted at an appropriate angle by rotating the angle adjustment knob 2124 to ensure that the tip of the first probe 216 can touch the bottom of the vessel 30 .

As shown in FIG. 5 , the microsphere lens 50 is fixed on the end of the probe (ie, the second probe 226 ) of the second micromanipulator 22 by glue. In this embodiment, microsphere lens 50 is a barium titanate glass lens. The resolution of traditional optical microscopes is limited by the diffraction limit, and the resolution can only reach half of the wavelength of the light source; the micron-sized silicon dioxide or barium titanate glass microsphere lens is placed on the surface of the sample to be observed, and can be observed through the microscope To the submicroscopic structure, it effectively breaks through the limitation of the diffraction limit and realizes super-resolution imaging. Since cells are dispersed in aqueous solution, the barium titanate glass lens is suitable for imaging in submerged or semi-submerged liquids. The microsphere lens is bonded to the tip of the probe by light-curing glue, and the probe is bonded to the upper half of the microsphere, which can ensure that the microsphere lens can fully contact the cells. As a tool for micro-manipulation of cells, the microsphere lens 50 can not only pick up the cells, but also use the magnification effect of the microsphere lens 50 to realize magnified observation of the cells and distinguish finer surface features.

In this embodiment, the first micro-manipulation manipulator 21 performs a first operation, and the first operation includes a cell puncture operation, a cell removal operation and/or a cell removal operation. The probe end of the second micro-manipulation manipulator 22 is provided with a microsphere lens 50 to perform the second operation. The second operation includes bonding and picking up cells. As known to those skilled in the art, in other embodiments, since the structure of the first micro-manipulation manipulator 21 is the same as that of the second micro-manipulation manipulator 22, the microsphere lens 50 can also be arranged on the first micro-manipulation manipulator 21, Then the first micro-manipulation manipulator 21 performs the second operation, and the second micro-manipulation manipulator 22 performs the first operation.

As shown in Fig. 6(a) and Fig. 6(b), the second operation includes bonding and picking up cells. Move the prepared microsphere probe to the bottom of the vessel 30, move the second micro-manipulator and the loading platform 40 below the vessel 30, select the cells to be picked up, and use the adsorption of water, the microsphere lens 50 can be bonded small number of cells. The cells attached to the microsphere lens 50 are moved to the middle of the liquid, away from the bottom of the vessel 30 .

As shown in FIGS. 7( a ), 7 ( b ) and 7 ( c ), the first operation includes the operation of puncturing cells, removing cells and/or clearing cells. Adjust the microscope to observe the morphology of the cells through the microsphere lens 50 in situ. Simultaneously, the first probe 216 of the first micro-operation manipulator 21 can be used to remove the cells at the bottom of the microsphere lens 50, so that the screening of the cells can be completed; The needle 216 can stab the cells at the bottom of the microsphere lens 50 to complete the observation of different cell shapes; at the same time, the first probe 216 of the first micromanipulator 21 can also be used to clean all the cells at the bottom of the microsphere lens 50 clean so that the microsphere probe 50 can be reused.

The in-situ cell observation and manipulation device based on the microsphere lens provided by the embodiment of the present invention uses the microsphere lens as a tool for cell micromanipulation, and realizes picking up cells with almost no damage and no shape change, and conveniently realizes sample sampling , and get rid of the shackles of the substrate in the prior art.

The cell in situ observation and manipulation device based on the microsphere lens utilizes the microsphere lens as a tool for cell micromanipulation, providing sufficient space for the manipulated cells to be observed.

The cell in situ observation and manipulation device based on the microsphere lens uses the microsphere lens as a tool for cell micromanipulation. The microsphere lens can realize super-resolution imaging of the imaging sample under the microscope, and can observe the cells more finely.

The cell in situ observation and manipulation device based on the microsphere lens uses the microsphere lens as a tool for cell micromanipulation. Through the microsphere lens, the shape of the cell can be observed in real time, and then the first micromanipulator can be used to realize the cell in situ. Convenient screening, puncturing, rejecting and other operations.

It will be apparent to those skilled in the art that the invention is not limited to the details of the above-described exemplary embodiments, but that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Accordingly, the embodiments should be regarded in all points of view as exemplary and not restrictive, the scope of the invention being defined by the appended claims rather than the foregoing description, and it is therefore intended that the scope of the invention be defined by the appended claims rather than by the foregoing description. All changes within the meaning and range of equivalents of the elements are embraced in the present invention. Any reference sign in a claim should not be construed as limiting the claim concerned.

In addition, it should be understood that although this specification is described according to implementation modes, not each implementation mode only contains an independent technical solution, and this description in the specification is only for clarity, and those skilled in the art should take the specification as a whole , the technical solutions in the various embodiments can also be properly combined to form other implementations that can be understood by those skilled in the art.

Claims (10)

1. A cell in situ observation and manipulation device based on a microsphere lens, characterized in that the cell in situ observation and manipulation device comprises: Microscope module, used to magnify cells to a preset magnification; an object loading platform, located below the eyepiece of the microscope module, for placing vessels; The first micro-manipulator is located on the first side of the microscope module, and is used to perform a first operation on the cells in the vessel; The second micromanipulator is located on the second side of the microscope module, and the first side and the second side are symmetrical about the central axis of the microscope module; The microsphere lens is arranged at the end of the probe of the second micro-manipulation manipulator, and is used for performing the second operation on the cells in the vessel. 2 . The device for observing and operating cells in situ based on microsphere lenses according to claim 1 , wherein the object loading platform has degrees of freedom in two directions. 3 . 3 . The in situ cell observation and manipulation device based on microsphere lenses according to claim 2 , wherein the object loading platform can move along the X-axis direction and along the Y-axis direction respectively. 4 . 4 . The in situ cell observation and manipulation device based on microsphere lenses according to claim 1 , wherein the structure of the first micro-manipulation manipulator is the same as that of the second micro-manipulation manipulator. 5. A kind of cell in-situ observation and operation device based on microsphere lens according to claim 1, characterized in that, the first micro-operation manipulator comprises: Probes, components for performing cellular manipulations; A probe extension rod, one end of the probe extension rod is fixedly connected to the probe, for extending the length of the probe; An angle adjustment mechanism, fixedly connected with the other end of the probe extension rod, for adjusting the angle of the probe; The main body is fixedly connected with the angle adjustment mechanism; a first mobile platform, fixedly connected to the main body, for adjusting the position of the probe in the first direction; a second mobile platform, connected to the first mobile platform, for adjusting the position of the probe in the second direction; The third mobile platform is connected with the second mobile platform and is used to adjust the position of the probe in the third direction. 6 . The device for in situ observation and manipulation of cells based on a microsphere lens according to claim 5 , wherein the first direction, the second direction and the third direction are respectively perpendicular to each other. 6 . 7 . The in-situ cell observation and manipulation device based on microsphere lenses according to claim 1 , wherein the microsphere lenses are fixed on the probe end of the second micromanipulator by glue. 8 . 8 . The device for in situ observation and manipulation of cells based on a microsphere lens according to claim 1 , wherein the first operation includes puncturing cells, screening cells and/or removing cells. 9 . 9 . The device for in situ observation and manipulation of cells based on microsphere lenses according to claim 1 , wherein the second operation includes bonding and picking up cells. 10 . 10 . The device for in situ observation and manipulation of cells based on a microsphere lens according to claim 1 , wherein the microsphere lens is a barium titanate glass lens. 11 .