阅读说明：本技术 一种用于太赫兹波辐照的贴壁细胞培养器 (Adherent cell culture device for terahertz wave irradiation ) 是由 周欢 彭晓昱 于 2021-07-19 设计创作，主要内容包括：本发明公开了一种用于太赫兹波辐照的贴壁细胞培养器,包括用于容纳细胞培养液并培养细胞的柱形腔室,柱形腔室底壁能够供太赫兹波聚焦波束穿过,培养细胞的环境为37℃、5％CO2、饱和湿度的细胞生长微环境,在柱形腔室内设置有凸起部,凸起部底面呈圆形并与柱形腔室底壁固定、重合并呈密封连接,位于凸起部侧壁与柱形腔室内壁之间的柱形腔室底壁作为细胞的环形贴壁生长面。本发明主要以结构简单的贴壁细胞培养器巧妙地解决了细胞太赫兹波生物效应一致性差的问题。(The invention discloses an adherent cell culture device for terahertz wave irradiation, which comprises a cylindrical chamber for containing a cell culture solution and culturing cells, wherein the bottom wall of the cylindrical chamber can be penetrated by a terahertz wave focused beam, the environment for culturing the cells is a cell growth microenvironment with 37 ℃, 5% CO2 and saturated humidity, a bulge part is arranged in the cylindrical chamber, the bottom surface of the bulge part is circular and is fixed with the bottom wall of the cylindrical chamber, the bulge part is overlapped and hermetically connected with the bottom wall of the cylindrical chamber, and the bottom wall of the cylindrical chamber positioned between the side wall of the bulge part and the inner wall of the cylindrical chamber is used as an annular adherent growth surface of the cells. The invention skillfully solves the problem of poor consistency of the terahertz wave biological effect of the cells by mainly using the adherent cell culture device with a simple structure.)

1. The utility model provides an adherent cell culture ware for terahertz wave irradiation, is including being used for holding cell culture solution and cultivateing cylindrical cavity (2) of cell, and cylindrical cavity (2) diapire can supply terahertz wave focusing wave speed to pass, cultivates the cell environment be 37 ℃, 5% CO2, the cell growth microenvironment of saturation humidity, its characterized in that: be provided with bellying (3) in cylindricality chamber (2), bellying (3) bottom surface is circular and fixed, coincide and be sealing connection with cylindricality chamber (2) diapire, and cylindricality chamber (2) diapire that is located between bellying (3) lateral wall and cylindricality chamber (2) inner wall is as the annular adherence growth surface of cell.

2. The adherent cell culture device for terahertz wave irradiation of claim 1, wherein: the bottom wall of the cylindrical chamber (2) is a hard polystyrene plastic sheet with the thickness not more than 0.5mm after surface modification treatment.

3. The adherent cell culture device for terahertz wave irradiation of claim 2, wherein: the annular adherent growth surface of the cell is completely projected in the radial section range of the strong irradiation; the terahertz focusing light velocity is generated by a double-color-field laser air plasma terahertz wave irradiation system and is focused by a necessary optical instrument.

4. The adherent cell culture device for terahertz wave irradiation of claim 3, wherein: the energy value of the strong irradiation is greater than 1/2 of the maximum value of the energy of the focused wave velocity of the terahertz waves.

5. The adherent cell culture device for terahertz wave irradiation of any one of claims 1 to 4, wherein: the adherent cell culture device comprises a cylindrical shell (7), the cylindrical chamber (2) is axially arranged on the shell (7), the shell (7) is clamped in a through hole (8) of a pore plate (9), and the pore plate (9) is fixed on an irradiation support; the convex part (3) is cylindrical or conical.

6. The adherent cell culture device for terahertz wave irradiation of any one of claims 1 to 4, wherein: the adherent cell culture device comprises a cylindrical shell (7), the cylindrical chamber (2) is axially arranged on the shell (7), and the shell (7) is directly fixed on the irradiation support; the boss (3) is composed of a cylindrical portion (32) and a conical portion (31) which are integrally formed and coaxially arranged.

7. The adherent cell culture device for terahertz wave irradiation of claim 6, wherein: a through hole is formed in the shell (7), and an arc-shaped ring groove (11) is formed in the inner side wall of the shell (7) close to the bottom of the through hole; conical part (31) bottom is provided with plectane (10) that are fit for the cell adherent growth, is provided with elasticity annular sealing element (12) at plectane (10) border fixed, and this elasticity annular sealing element (12) can imbed in arc annular (11), by plectane (10) conical part (31) lateral wall with casing (7) inside wall encloses synthetic splendid attire cell culture liquid's space jointly, plectane (10) surface does cylindricality cavity (2) diapire.

8. The adherent cell culture device for terahertz wave irradiation of any one of claims 1 to 4, wherein: the adherent cell culture device comprises a cylindrical shell (7), the cylindrical chamber (2) is axially arranged on the shell (7), and the shell (7) is directly fixed on the irradiation support; the boss (3) is cylindrical, and a support lug (33) is arranged on the top surface of the boss (3); a through hole is formed in the shell (7), and an arc-shaped ring groove (11) is formed in the inner side wall of the shell (7) close to the bottom of the through hole; bellying (3) bottom is provided with plectane (10) that are fit for the cell adherent growth, is provided with elasticity annular sealing element (12) at plectane (10) border fixed, and this elasticity annular sealing element (12) can imbed in arc annular (11), by plectane (10) bellying (3) lateral wall with casing (7) inside wall encloses the space of synthetic splendid attire cell culture liquid jointly, plectane (10) surface does cylindricality chamber (2) diapire.

9. The adherent cell culture device for terahertz wave irradiation of claim 1, wherein: the bottom wall of the cylindrical chamber (2) adopts a horizontal hard polystyrene plastic film with the thickness not more than 0.2mm after surface modification treatment, and the bulge (3) adopts a capsule filled with liquid and fully expanded; the bottom wall of the cylindrical cavity (2) is connected with the bottom surface of the bulge (3) in a sealing bonding mode.

10. The adherent cell culture device for terahertz wave irradiation of any one of claims 2 to 9, wherein: the bulge (3) and the shell (7) are both made of hydrophobic polystyrene materials.

Technical Field

The invention belongs to the technical field of terahertz wave biological effects, and particularly relates to an adherent cell culture device for terahertz wave irradiation.

Background

In recent years, the application of terahertz technology is increasingly widespread, and the application research of imaging and sensing technology in the aspects of medical treatment, military affairs and security is continuously intensive. In order to safely and effectively apply the terahertz technology, particularly to the fields of diagnosis, living body imaging, treatment, security inspection and the like, the biological effect and the safety of terahertz radiation need to be researched and evaluated, the biological effect characteristic and the dose-effect relationship are determined, and the injury threshold is obtained. Therefore, the research on the biological effect of terahertz waves becomes a focus of great attention in the field.

In the prior art, document CN111239377A discloses a system and a method for researching the biological effect of terahertz waves of cells isolated from human bodies for a long time, which can realize the long-time in vitro irradiation of terahertz waves on various primary cells and cell lines growing adherent to the human bodies, and provide a stable experimental platform and a stable method for researching the biological effect of terahertz waves. The system and the method only ensure the growth environment of the cells during in vitro irradiation from the constant-temperature sample chamber, namely the temperature, the humidity and the carbon dioxide concentration are all consistent with the incubator environment, thereby eliminating the possible effect of the change of the external environment on the growth state of the cells. However, researches show that the factors influencing the consistency of the cellular effect under the terahertz radiation are not only growth environments, but also the problem of poor consistency of the biological effect of the terahertz waves of the cells still exists by adopting the system and the method.

Disclosure of Invention

The invention aims to provide an adherent cell culture device for terahertz wave irradiation, which is at least used for solving the problem of poor consistency of biological effects of terahertz waves of cells.

The purpose of the invention is realized by adopting the following technical scheme.

The utility model provides an adherent cell culture ware for terahertz wave irradiation, is including the cylindricality chamber that is used for holding cell culture liquid and cultivates the cell, and cylindricality chamber diapire can supply terahertz wave focusing beam to pass, cultivates the cell environment be 37 ℃, 5% CO2, the cell growth microenvironment of saturation humidity, its characterized in that: the cylindrical chamber is internally provided with a bulge, the bottom surface of the bulge is circular and is fixed with the bottom wall of the cylindrical chamber, overlapped and hermetically connected, and the bottom wall of the cylindrical chamber between the side wall of the bulge and the inner wall of the cylindrical chamber is used as an annular adherent growth surface of cells.

In order to improve the stability of the terahertz wave irradiation process, the bottom wall of the cylindrical cavity is a hard polystyrene plastic thin plate with the thickness not greater than 0.5mm after surface modification treatment.

In order to further improve the biological effect consistency of the cell terahertz waves, the cell adherent growth surface is completely projected in the radial section range of the strong irradiation position by controlling the diameter of the bulge part, namely the width H of the cell adherent growth surface is smaller than the width of the radial section of the strong irradiation position, and the terahertz focused beam is generated by a bicolor field laser air plasma terahertz wave irradiation system and is focused by a necessary optical instrument. The necessary optics include, but are not limited to, off-axis parabolic mirrors, THz total reflection lenses, THz lenses.

Preferably, the energy value of the strong irradiation is greater than 1/2 of the energy maximum value of the terahertz wave focused beam.

For convenient operation, the adherent cell culture device comprises a cylindrical shell, the cylindrical cavity is axially arranged on the shell, the shell is used for being clamped in a through hole of a pore plate, and the pore plate is fixed on an irradiation support; the convex part is cylindrical or conical.

Furthermore, the adherent cell culture device comprises a cylindrical shell, the cylindrical chamber is axially arranged on the shell, and the shell is directly fixed on the irradiation support; the convex part is composed of a cylindrical part and a conical part which are integrally formed and coaxially arranged.

As a preferred embodiment of the invention, the shell is provided with a through hole, and the inner side wall of the shell close to the bottom of the through hole is provided with an arc-shaped annular groove; the cone bottom is provided with the plectane that is fit for the cell adherent growth, is provided with elasticity annular sealing element at the plectane border is fixed, and this elasticity annular sealing element can imbed in the arc annular, by the plectane the cone lateral wall with the space of synthetic splendid attire cell culture liquid is enclosed jointly to the casing inside wall, the plectane surface does the cylindricality cavity diapire. Adopt such structure can also improve the casing utilization ratio, be favorable to practicing thrift the consumptive material. In the invention, the elastic annular sealing elements are all made of materials meeting medical requirements.

As another preferred embodiment of the present invention, the adherent cell culture vessel comprises a cylindrical housing, the cylindrical chamber is axially disposed on the housing, and the housing is directly fixed on the irradiation support; the lug boss is cylindrical, and a support lug is arranged on the top surface of the lug boss; the shell is provided with a through hole, and the inner side wall of the shell close to the bottom of the through hole is provided with an arc-shaped ring groove; the bellying bottom is provided with the plectane that is fit for the cell adherent growth, is provided with elasticity annular sealing element at the plectane border is fixed, and this elasticity annular sealing element can imbed in the arc annular, by the plectane the bellying lateral wall with the space of synthetic splendid attire cell culture liquid is enclosed jointly to the casing inside wall, the plectane surface does the cylindricality chamber diapire. By adopting the structure, the cell culture auxiliary operation is facilitated, for example, the circular plate for cell adherent growth is disassembled and assembled, and the growth condition of the irradiated cells after terahertz irradiation is conveniently observed.

In order to further improve the stability in the terahertz wave irradiation process, the bottom wall of the cylindrical cavity adopts a horizontal hard polystyrene plastic film with the thickness not more than 0.2mm after surface modification treatment, and the bulge adopts a capsule filled with liquid and fully expanded; the bottom wall of the cylindrical cavity is connected with the bottom surface of the bulge in a sealing bonding mode.

In order to further improve the stability of the terahertz wave in the irradiation process, the bulge and the shell are made of hydrophobic polystyrene materials, the scheme can prevent cells from growing by being attached to the side wall of the bulge, and errors of the consistency evaluation of the terahertz wave biological effect of the irradiated cells are reduced.

Has the advantages that: the invention skillfully solves the problems of poor uniformity of irradiated cells and poor consistency of terahertz wave biological effect mainly by using the adherent cell culture device with a simple structure, and when the terahertz wave irradiation is implemented, the specific structure of the adherent cell culture device is used for avoiding the terahertz spectrum/light wave energy which does not meet the energy requirement, thereby greatly improving the consistency of the terahertz wave biological effect of the irradiated cells; the adherent cell culture device for terahertz wave irradiation provided by the invention is beneficial to improving the uniformity and stability in the terahertz wave irradiation process and the convenience of operation, and common technicians in the field can smoothly implement the operation; the adherent cell culture device for terahertz wave irradiation provided by the invention can be used for repeatedly observing adherent cells of the same batch at intervals without changing a cell culture vessel, and is favorable for improving the accuracy and efficiency of experimental results.

Drawings

FIG. 1 is a schematic sectional view of an adherent cell culture vessel for terahertz wave irradiation in example 1;

FIG. 2 is a schematic view of a circular plate and a convex portion of the adherent cell culture vessel for terahertz wave irradiation in example 1;

FIG. 3 is a schematic sectional view of an adherent cell culture vessel for terahertz wave irradiation in example 2;

FIG. 4 is a schematic view of a circular plate and a convex portion of the adherent cell culture vessel for terahertz wave irradiation in example 2;

FIG. 5 is a schematic view of an adherent cell culture vessel for terahertz wave irradiation in example 3;

FIG. 6 is a schematic view of an orifice plate in example 3;

FIG. 7 is a schematic view of an adherent cell culture vessel for terahertz wave irradiation in example 4;

FIG. 8 is a schematic view of an adherent cell culture vessel for terahertz wave irradiation in example 5;

FIG. 9 is a partial schematic view of a terahertz wave irradiation optical path in embodiments 1 to 5;

FIG. 10 is a schematic diagram of a light spot focused by the terahertz wave irradiation optical path in the embodiment 1-5;

h in FIG. 1 represents the width H of the cell adherent growth surface, and H in FIG. 9 represents the width/diameter of the weakly irradiated region.

Detailed Description

The present invention is further illustrated by the following examples, which are not to be construed as limiting the scope of the invention, and that modifications and variations that are not essential to the invention may be made by those skilled in the art in light of the teachings herein.

Example 1

Referring to fig. 1 and 2, an adherent cell culture device for terahertz wave irradiation comprises a cylindrical shell 7, wherein the shell 7 is directly fixed on an irradiation support; the cylindrical chamber 2 is axially arranged on the shell 7, the cylindrical chamber 2 is a part of a through hole on the shell 7 substantially, the cylindrical bulge 3 is arranged in the cylindrical chamber 2, the top surface of the bulge 3 is provided with a support lug 33, the support lug 33 is mainly used for clamping tools such as a clamp or a tweezers, the bottom surface of the bulge 3 is circular and is fixed with the bottom wall of the cylindrical chamber 2, the bottom wall of the cylindrical chamber 2 between the side wall of the bulge 3 and the inner wall of the cylindrical chamber 2 is superposed and connected in a sealing way, and the bottom wall of the cylindrical chamber 2 serves as an annular wall-attached growth surface of cells. An arc-shaped annular groove 11 is formed in the inner side wall of the shell 7 close to the bottom of the through hole, a circular plate 10 suitable for adherent growth of cells is arranged at the bottom of the boss 3, an elastic annular sealing element 12 is fixedly arranged at the edge of the circular plate 10, the elastic annular sealing element 12 can be embedded into the arc-shaped annular groove 11, a space for containing cell culture solution is formed by the circular plate 10, the side wall of the boss 3 and the inner side wall of the shell 7 in a surrounding mode, the surface of the circular plate 10 is the bottom wall of the cylindrical cavity 2, the bottom wall (namely the circular plate 10) of the cylindrical cavity 2 can be penetrated by terahertz focused beams, and the terahertz focused beams can penetrate through the circular plate 10 and irradiate onto the adherent cells to generate a biological effect.

The bottom wall (circular plate 10) of the cylindrical chamber 2 is a hard polystyrene plastic thin plate with the thickness not greater than 0.3mm after surface modification treatment, the hard polystyrene plastic thin plate after surface modification treatment is favorable for adherent growth of cells, the bulge part 3 and the shell 7 are both made of hydrophobic polystyrene materials, and the hydrophobic polystyrene materials can prevent adherent growth of the cells on the side wall of the bulge part 3 and the side wall of the shell 7.

The principle of terahertz wave irradiation of adherent cells in this embodiment: a terahertz wave irradiation optical path 21 which is generated by a double-color field laser air plasma terahertz irradiation system and focused by a necessary optical instrument is distributed in a conical shape, as shown in fig. 9 and 10, along the radial section of the conical shape, the photon energy close to an axis 26 is weaker, the photon energy of the part is difficult to meet the irradiation requirement on adherent cells, a peak trough indicated by a number 25 in fig. 9 represents the energy intensity of a terahertz wave focused beam, a peak area 22 (namely a strong irradiation area, the energy value of the area is larger than 1/2 of the energy maximum value of the terahertz focused beam) has stronger energy of the terahertz spectrum/light wave, a trough area 23 (namely a weak irradiation area, the energy value of the area is smaller than 1/2 of the energy maximum value of the terahertz wave focused beam) has weaker energy of the terahertz spectrum/light wave, and the conical top part is a light spot 24 after the terahertz wave irradiation optical path is focused, the weaker energy in the central region of spot 24 corresponds to valley regions 23 and the stronger energy in regions outside the central region of spot 24 corresponds to peak regions 22. In this embodiment, the circular plate 10 is provided with the protruding portion 3, so that the cells can only perform adherent growth on the bottom wall of the cylindrical chamber 2 between the side wall of the protruding portion 3 and the inner wall of the cylindrical chamber 2, and the adherent growth surface of the cells is completely projected in the radial cross section range of the strong irradiation place by controlling the diameter of the protruding portion 3, that is, the width H of the adherent growth surface of the cells is smaller than the width of the radial cross section of the strong irradiation place, and the adherent growth area of the cells just corresponds to the peak area 22 with stronger energy, so that the terahertz spectrum/light wave energy which does not meet the energy requirement is ingeniously avoided, and the uniformity of the irradiated cells and the consistency of the terahertz wave biological effect are greatly improved.

Before terahertz wave irradiation is carried out, a circular plate 10 with a protruding part 3 is clamped in an arc-shaped annular groove 11 in a cylindrical chamber 2, a cell growth microenvironment with 37 ℃, 5% CO2 and saturated humidity is adopted as an environment for culturing cells, after the cells in the cylindrical chamber 2 grow to a proper stage in an adherent mode, the cells are transferred to a microscopic device for observation or sampled and observed under a glass slide (under a pollution-free environment usually), and the cells are continuously cultured in the next stage after observation. Because the edge of the circular plate 10 is fixedly provided with the elastic annular sealing element 12, when the circular plate 10 needs to be taken out under special conditions, the elastic annular sealing element 12 is still attached to the circular plate 10 after the circular plate 10 is detached, so that a space for storing cell growth liquid is provided for cells attached to the circular plate 10, cell growth is facilitated in an observation stage, multiple repeated observation at intervals can be performed on the attached cells of the same batch under the condition that a cell culture vessel is not replaced, and the accuracy and the efficiency of an experimental result are improved. Moreover, the process of assembling and disassembling the circular plate 10 is simple and convenient, and the operation can be smoothly carried out by a person skilled in the art.

Example 2

An adherent cell culture device for terahertz wave irradiation, referring to example 1 and shown in fig. 3 and 4, the main difference from example 1 is the structure of the projection part 3, and the circular plate 10 of the adherent cell culture device is easier to disassemble compared with example 1: the adherent cell culture device comprises a cylindrical shell 7, a cylindrical chamber 2 is arranged on the shell 7, and the shell 7 is directly fixed on an irradiation bracket; the boss 3 is composed of a cylindrical part 32 and a conical part 31 which are integrally formed and coaxially arranged, and the cylindrical part 32 is convenient to hold in the assembling or disassembling process; a through hole is formed in the shell 7, and an arc-shaped annular groove 11 is formed in the inner side wall of the shell 7 close to the bottom of the through hole; a circular plate 10 suitable for the adherent growth of cells is arranged at the bottom of the conical part 31, an elastic annular sealing element 12 is fixedly arranged at the edge of the circular plate 10, the elastic annular sealing element 12 can be embedded into the arc-shaped annular groove 11, a space for containing cell culture solution is jointly enclosed by the circular plate 10, the side wall of the conical part 31 and the inner side wall of the shell 7, and the surface of the circular plate 10 is the bottom wall of the cylindrical chamber 2.

Example 3

An adherent cell culture device for terahertz wave irradiation is disclosed, as shown in fig. 5 and 6, and comprises a shell 7, wherein a support lug 5 which is convenient for holding a pinching part is arranged on the shell 7, the shell 7 is used for being clamped in a through hole 8 of an orifice plate 9, the orifice plate 9 is fixed on an irradiation support, a cylindrical chamber 2 which is arranged on the shell 7 and used for containing cell culture solution and culturing cells is arranged, the bottom wall of the cylindrical chamber 2 can be penetrated by a terahertz focused beam, and the environment for culturing the cells is a cell growth microenvironment with 37 ℃, 5% CO2 and saturated humidity; be provided with bellying 3 in cylindricality chamber 2, bellying 3 is cylindrically, and 3 bottom surfaces of bellying are circular and fixed, coincide and be sealing connection with 2 diapalls of cylindricality chamber, and 2 diapalls of cylindricality chamber that lie in between 3 lateral walls of bellying and the 2 inner walls of cylindricality chamber are as the annular adherence growth surface of cell.

The bottom wall of the cylindrical chamber 2 is a hard polystyrene plastic sheet with the thickness not greater than 0.3mm after surface modification treatment, the hard polystyrene plastic sheet after surface modification treatment is favorable for adherent growth of cells, the bulge 3 and the shell 7 are both made of hydrophobic polystyrene materials, and the hydrophobic polystyrene materials can prevent adherent growth of the cells on the side walls of the bulge 3 and the shell 7.

The principle of terahertz wave irradiation of adherent cells in this example is similar to that of example 1. Other advantages of the adherent cell culture apparatus of the present embodiment include mainly: conveniently get and put a single subassembly 6 that has cell culture liquid and bellying 3, orifice plate 9 can reuse, is favorable to practicing thrift the orifice plate.

Example 4

An adherent cell culture device for terahertz wave irradiation, as shown in reference to example 3 in combination with fig. 7, differs from example 1 mainly in the structure of a convex portion 3: the boss 3 is conical and the apex of the boss is lower than the top of the housing 7.

Example 5

An adherent cell culture device for terahertz wave irradiation, as shown in reference to example 3 in combination with fig. 8, differs from example 1 mainly in the structure of a convex portion 3: the boss 3 is conical and the boss apex is flush with the top of the housing 7.

In addition, a cover is provided on the top of the housing 7, and the cover is covered during the cell culture process to prevent the environment in the cylindrical chamber 2 from being contaminated.