KR102639623B1 - Time-lapse cell imaging apparatus - Google Patents

Abstract

The present invention relates to a time-lapse culture cell imaging device that can monitor a culture dish containing cells by photographing them directly from inside the chamber without taking them out of the chamber of the culture device. The time-lapse culture cell imaging device according to the present invention is a A case in which an opening is formed on the surface to be open upward and downward; A mount member disposed inside the opening of the case and on which a culture dish for cultivating cells is mounted; An X-Y-Z stage installed below the mount member inside the case to move the mount member in the X, Y, and Z axes; an imaging unit installed inside the case to penetrate the X-Y-Z stage and having an objective lens to photograph cells inside the culture dish through the lower side of the mount member; a lighting unit installed outside the case to emit light for photography toward the petri dish; And, an airtight maintenance portion that seals the space between the inner periphery of the opening of the case and the outer surface of the mount member while supporting the mount member to be movable in the X, Y, and Z axes.

Description

Time-lapse culture cell imaging device {TIME-LAPSE CELL IMAGING APPARATUS}

The present invention relates to a device for photographing cells in a culture dish, and more specifically, to monitor the culture state of cells by moving a culture dish with a plurality of wells for culturing cells in the horizontal and vertical directions. It relates to a time-lapse cultured cell imaging device.

Cell culture is a very important technology in biological research, including molecular biology, and refers to cultivating specific cells for purposes such as diagnosis or treatment of human diseases. Generally, cell culture is performed using a cell imaging device, and a conventional cell imaging device includes a cover glass, a culture dish, and a cover.

The method of culturing and observing cells using a conventional cell imaging device is as follows. Cells treated with drugs or cells with recombinant specific genes and culture medium are placed on a thin glass cover glass, placed in a culture dish, covered with a cover, and the covered culture dish is placed in an incubator, thereby forming cells. Complete preparations for culture. The incubator proliferates cells by creating a specific environment suitable for the purpose of cell culture. Once cell proliferation is completed, the cover glass is taken out with tweezers, a glass slide is placed on it, and the cells are observed through a microscope. However, this conventional method not only has the inconvenience of having to separately prepare a cover glass, a culture dish, and a cover, but also has the problem that there is a high possibility that cells will be contaminated during the cell culture preparation process.

Republic of Korea Patent Publication No. 10-2019-0080747 discloses a time-lapse cell imaging device that controls an environment suitable for cell culture and periodically photographs the cell culture process.

In the time-lapse cell imaging device of the above-mentioned published patent, the culture dish containing the cells is fixed, and the imaging unit for photographing cells is moved in the It is configured for monitoring.

The time-lapse cell imaging device of the published patent has the advantage of improving the reliability and efficiency of cell imaging without shaking or entangling the cells by moving the imaging unit without moving the culture dish.

However, conventional cell imaging devices, including the time-lapse cell imaging device of the above published patent, are installed outside the culture chamber where cell culture is performed, so cells are contaminated during the process of taking the culture dish out of the culture chamber and transferring it to the imaging device and imaging. There is always a possibility that you will become stressed or become severely stressed.

Accordingly, there is a demand for the development of a cell imaging device that can be installed inside a culture chamber to capture cells in a controlled environment inside the chamber without transporting the culture dish to the outside of the culture chamber.

However, the time-lapse cell imaging device of the published patent has a problem in that it is difficult to install inside a culture chamber because the configuration for moving the imaging unit in the X, Y, and Z axes is complicated and large.

In addition, since the inside of the culture chamber is an environment where the humidity is controlled to be somewhat high for cell culture, when an imaging device is installed inside the culture chamber, the lens of the imaging unit, the drive motor for moving the imaging unit, and electronic components must be sealed in an airtight space. However, as the size of the device becomes larger and more complex, the configuration for maintaining confidentiality becomes more complex, and maintaining confidentiality becomes difficult.

Republic of Korea Patent Publication No. 10-2019-0080747

The present invention is intended to solve the above-described problem, and the purpose of the present invention is to monitor the culture dish containing the cells by taking pictures directly from inside the chamber without taking them out of the chamber of the culture device, and to provide a configuration for taking pictures and moving the position. The aim is to provide a time-lapse culture cell imaging device that has a simple configuration of elements and can ensure confidentiality.

A time-lapse cultured cell imaging device according to the present invention for achieving the above object includes a case having an opening on the upper surface that opens upward and downward; A mount member disposed inside the opening of the case and on which a culture dish for cultivating cells is mounted; An X-Y-Z stage installed below the mount member inside the case to move the mount member in the X, Y, and Z axes; an imaging unit installed inside the case to penetrate the X-Y-Z stage and having an objective lens to photograph cells inside the culture dish through the lower side of the mount member; a lighting unit installed outside the case to emit light for photography toward the petri dish; And, an airtight maintenance portion that seals the space between the inner periphery of the opening of the case and the outer surface of the mount member while supporting the mount member to be movable in the X, Y, and Z axes.

The mounting member may be formed in the form of a cylinder open up and down.

According to one form of the present invention, the airtightness maintenance unit includes a ring-shaped seal housing fixed to the inner periphery of the opening; an outer fixing ring in the form of a ring fixed to the upper side of the seal housing at regular intervals; And, an inner side that is in the form of a ring and is horizontally slidable between the outer fixing rings while maintaining airtightness with respect to the outer fixing ring, and is in close contact with the inner peripheral portion so that the outer surface of the mount member can slide up and down while maintaining airtightness. It may include a sliding ring.

The seal housing includes a first seal housing in the form of a ring that is fixed to a lower side of the inner periphery of the opening and has an 'L'-shaped cross section on which an inner slide ring disposed at the lowermost side of the inner slide rings is slidably seated, It may include a second chamber housing that is installed to extend upward from the first chamber housing and supports the outer fixing ring.

A time-lapse cultured cell imaging device according to another form of the present invention includes a ring-shaped coupling portion that is in close contact with the upper outer surface of the mount member on the outside of the case, the coupling portion being formed to extend radially, and an outer peripheral portion. It may further include an external airtight cover including a plane movable airtight portion that is connected to the upper surface of the case and seals between the top of the mount member and the periphery of the opening.

A time-lapse cultured cell imaging device according to another form of the present invention is connected to the outer surface of the coupling portion of the external airtight cover at the upper end of the mount member, while maintaining airtightness with respect to the outer surface of the coupling portion when the mount member moves up and down. It may further include a sealing flange that slides up and down.

According to the present invention, when the electrical components such as the imaging unit, It can be supported while moving smoothly in the X-axis, Y-axis, and Z-axis directions by the holding part.

Therefore, with the imaging unit and lighting unit fixed without moving in the Miniaturization is possible by significantly reducing the overall size of the device.

In this way, airtightness can be continuously maintained when photographing cultured cells by moving the mount member, and miniaturization is possible, so the cell imaging device can be easily installed inside the chamber of the culture device, rather than transporting the culture dish outside the chamber. Since imaging can be done right away, contamination can be prevented during the process of transporting the culture dish to the outside, and cells can be prevented from being stressed.

Figure 1 is a perspective view showing a time-lapse cultured cell imaging device according to an embodiment of the present invention.
Figure 2 is a perspective view showing a cutaway time-lapse culture cell imaging device according to an embodiment of the present invention.
Figure 3 is a cross-sectional view of main parts of a time-lapse culture cell imaging device according to an embodiment of the present invention.
Figure 4 is an enlarged cross-sectional view of part A of Figure 3.

Hereinafter, the present invention will be described with reference to the attached drawings. However, the present invention may be implemented in various different forms and, therefore, is not limited to the embodiments described herein. In order to clearly explain the present invention in the drawings, parts that are not related to the description are omitted, and similar parts are given similar reference numerals throughout the specification.

Throughout the specification, when a part is said to be "connected (connected, contacted, combined)" with another part, this means not only "directly connected" but also "indirectly connected" with another member in between. "Includes cases where it is. Additionally, when a part is said to “include” a certain component, this does not mean that other components are excluded, but that other components can be added, unless specifically stated to the contrary.

The terms used herein are only used to describe specific embodiments and are not intended to limit the invention. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as “comprise” or “have” are intended to indicate the presence of features, numbers, steps, operations, components, parts, or combinations thereof described in the specification, but are not intended to indicate the presence of one or more other features. It should be understood that this does not exclude in advance the possibility of the existence or addition of elements, numbers, steps, operations, components, parts, or combinations thereof.

Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings.

1 to 4, the time-lapse culture cell imaging device according to an embodiment of the present invention includes a case 100, a mount member 500, an X-Y-Z stage 400, an imaging unit 200, and a lighting unit ( 300), and a confidentiality department (600).

The case 100 may have a rectangular box shape, and an opening 111, where the mount member 500 and the airtight maintenance portion 600 are installed, is formed on one side of the upper surface 110 to be open up and down. The opening 111 may be formed in a circular shape. Case 100 is installed inside the culture chamber of the culture device where the culture dish 1 is accommodated.

The mounting member 500 has a cylindrical shape open up and down, and is supported inside the opening 111 of the case 100 by the airtight maintenance portion 600 so as to be movable in the X, Y, and Z axes. A culture dish 1 for cultivating cells is seated and supported on the upper end of the mount member 500. The lower end of the mount member 500 is coupled to the X-Y-Z stage 400 and moves in the X, Y, and Z axes directions by the X-Y-Z stage 400.

The culture dish 1 mounted on the mount member 500 has a plurality of wells for culturing cells. The culture dish 1 is placed on the upper part of the mount member 500 when the culture state of cells is to be monitored, and when imaging is completed, it is separated from the upper part of the mount member 500 and transported to a designated location.

The X-Y-Z stage 400 is installed on the lower side of the mount member 500 inside the case 100 and moves the mount member 500 in the X, Y, and Z axes. The X-Y-Z stage 400 can be configured by applying a known X-Y-Z stage that can precisely move to the desired direction and position of the X, Y, and Z axes by a linear motion device including You can. Since the X-Y-Z stage is a well-known component in the art, detailed description of its composition and operation will be omitted.

The center of the X-Y-Z stage 400 is formed to be open, so that the objective lens 220 of the imaging unit 200 is disposed through the open center of the ) can be photographed.

The imaging unit 200 is fixedly installed inside the case 100. The imaging unit 200 includes an optical tube 210 installed at the end of an objective lens 220 for photographing cells inside the culture dish 1 through the open lower part of the mount member 500, and the optical tube 210. It may include a camera 230 installed on the other end. The image captured by the imaging unit 200 is transmitted to the control unit (not shown) of the imaging device.

The lighting unit 300 is fixedly installed on the outside of the upper surface 110 of the case 100 and emits light for photography toward the culture dish 1 seated on the top of the mount member 500. The lighting unit 300 operates by receiving power from a control unit (not shown) of the photographing device.

The imaging unit 200 and the lighting unit 300 can also be configured by applying known imaging devices and lighting devices included in general cell imaging devices.

Since the X-Y-Z stage 400 and the imaging unit 200 are components sensitive to moisture and contamination by foreign substances, it is desirable to install them inside the case 100 while maintaining airtightness.

The airtight maintenance unit 600 airtightly seals the space between the outer surface of the mount member 500 and the inner periphery of the opening 111 so that various components installed inside the case 100 can be maintained in a sealed state. It is a component that supports the movable mount member 500 so that it can move smoothly in the X, Y, and Z axes.

The airtight maintenance unit 600 may include a seal housing 610, a plurality of outer fixing rings 620, a plurality of inner slide rings 630, an external airtight cover 650, and a sealing flange 660. You can.

The seal housing 610 is in the form of a circular ring with an open center and is fixed to the lower side of the inner periphery of the opening 111. The first seal housing 611 has a ring shape with an approximately 'L' shaped cross section; , and includes a second chamber housing 612 that is installed to extend upward from the first chamber housing 611 and supports the plurality of outer fixing rings 620.

On the upper surface of the first chamber housing 611, an inner slide ring 630 disposed at the lowermost position among the plurality of inner slide rings 630 is slidably seated while maintaining airtightness.

The outer fixing ring 620 is in the form of a circular ring with an open center, and the outer end is installed on the inner peripheral surface of the second chamber housing 612 at regular intervals up and down. The outer diameter of the outer fixing ring 620 is formed to be almost the same as the inner diameter of the second chamber housing 612, so that the outer circumferential surface of the outer fixing ring 620 is fixedly supported by the inner circumferential surface of the second chamber housing 612. .

The inner slide ring 630 is in the form of a circular ring with an outer diameter smaller than the inner diameter of the second chamber housing 612 but larger than the inner diameter of the outer fixing ring 620, and is located between the outer fixing ring 620 and the bottom. are each slidably disposed between the outer fixing ring 620 and the upper surface of the first chamber housing 611. A plurality of outer fixing rings 620 and a plurality of inner slide rings 630 are alternately arranged.

The inner peripheral portion of the inner slide ring 630 is in close contact with the outer surface of the mount member 500. At this time, the outer surface of the mount member 500 is in close contact with the inner periphery of the inner slide ring 630 so that it can slide up and down while maintaining airtightness.

As the mount member 500 is supported in close contact with the inner periphery of the inner slide ring 630, when the mount member 500 moves in the horizontal direction by the X-Y-Z stage 400, the inner slide ring 630 moves to the outer side. Since it slides while maintaining airtightness with respect to the fixing ring 620 and the upper surface of the first chamber housing 611, the mount member 500 can move smoothly to the desired position and at the same time maintain airtightness.

In addition, even when the mount member 500 moves up and down by the You can.

Due to the relative movement and airtightness function of the seal housing 610, the plurality of outer fixing rings 620, and the plurality of inner slide rings 630, the mount member 500 is smooth and smooth in the It is possible to maintain high confidentiality while moving freely.

As the airtightness maintenance part 600, a bellows-type airtightness maintenance part may be applied, but the bellows-type airtightness maintenance part requires space for installation flanges at both ends and has a large degree of freedom in the axial direction, In the radial direction, a certain amount of acid is required due to deformation, which may limit miniaturization in the longitudinal direction. In addition, the operating resistance is highly dependent on the number of threads of the bellows, so in order to use it with a small driving force of a small stage, a bellows with a larger number of threads must be used.

In order to further improve the airtightness by the airtightness maintenance unit 600, an external airtight cover 650 and a sealing flange 660 that primarily exert airtightness may be formed on the outside of the case 100.

The external airtight cover 650 includes a ring-shaped coupling portion 651 that is in close contact with the upper outer surface of the mount member 500 on the outside of the upper surface 110 of the case 100, and an outer periphery of the lower end of the coupling portion 651. A plane movable airtight portion 652 is formed to extend radially at the edge, and the outer peripheral portion is connected to the upper surface 110 of the case 100 and seals the space between the upper outer surface of the mount member 500 and the peripheral portion of the opening 111. ) may include.

The outer peripheral edge of the plane movable airtight portion 652 is bent downward so that the distal end is connected to the upper surface 110 of the case 100, so that when the mount member 500 moves horizontally along the X-axis or Y-axis, the case It maintains airtightness while sliding along the upper surface 110 of (100). At this time, the lower surface of the flat movable airtight portion 652 slides while being in close contact with the upper surface of the uppermost outer fixing ring 620.

The sealing flange 660 is formed in an 'L' shape while extending radially outward at the upper end of the mount member 500, and is relative to the outer surface of the coupling portion 651 as the mount member 500 moves up and down. It functions to maintain airtightness as it moves up and down while sliding.

This cell imaging device is installed inside the case 100 by the airtight maintenance unit 600 with electrical components such as the imaging unit 200, X-Y-Z stage 400, and control unit (not shown) installed inside the case 100. In addition to maintaining airtightness, the mount member 500 on which the culture dish 1 is seated can be supported while moving smoothly in the X-axis, Y-axis, and Z-axis directions by the airtightness maintaining portion 600.

Therefore, while the imaging unit 200 and the lighting unit 300 are fixed without moving in the Since cultured cells can be photographed, the overall size of the cell imaging device can be greatly reduced and miniaturized, and the cell imaging device can be easily installed inside the chamber of the culture device where the culture dish (1) is accommodated. It is possible to film and monitor the culture status right inside the chamber without transporting it outside the chamber.

The description of the present invention described above is for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive. For example, each component described as single may be implemented in a distributed manner, and similarly, components described as distributed may also be implemented in a combined form.

The scope of the present invention is indicated by the patent claims described below, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be construed as being included in the scope of the present invention.

1: Petri dish 100: Case
200: imaging unit 220: objective lens
300: Lighting unit 400: XYZ stage
500: Mounting member 600: Confidentiality maintenance part
610: seal housing 620: outer fixing ring
630: Inner slide ring 650: External airtight cover
660: Sealing flange

Claims (6)

A case having an opening on the upper surface that opens upward and downward;
A mount member disposed inside the opening of the case and on which a culture dish for cultivating cells is mounted;
An XYZ stage installed below the mount member inside the case to move the mount member in the X, Y, and Z axes;
An imaging unit installed inside the case to penetrate the XYZ stage and having an objective lens to photograph cells inside the culture dish through the lower side of the mount member;
a lighting unit installed outside the case to emit light for photography toward the petri dish; and,
an airtight maintenance portion that seals the space between the inner periphery of the opening of the case and the outer surface of the mount member while supporting the mount member to be movable in the X, Y, and Z axes;
Including,
The confidentiality department said,
a ring-shaped seal housing fixed to the inner periphery of the opening;
an outer fixing ring in the form of a ring fixed to the upper side of the seal housing at regular intervals; and,
An inner slide ring is arranged in the form of a ring so that it can slide horizontally between the outer fixing rings while maintaining airtightness with respect to the outer fixing ring, and the outer surface of the mount member is in close contact with the inner periphery so that it can slide up and down while maintaining airtightness. ;
Includes,
The seal housing includes a first seal housing in the form of a ring that is fixed to a lower side of the inner periphery of the opening and has an 'L'-shaped cross section on which an inner slide ring disposed at the lowermost side of the inner slide rings is slidably seated, A time-lapse culture cell imaging device comprising a second chamber housing that is installed to extend upward to the first chamber housing and supports the outer fixing ring. The time-lapse culture cell imaging device according to claim 1, wherein the mount member is in the form of a cylinder open up and down. delete delete The mount according to claim 1, wherein a ring-shaped coupling part is in close contact with the upper outer surface of the mount member on the outside of the case, the coupling part is formed to extend radially, and an outer peripheral part is connected to the upper surface of the case. A time-lapse culture cell imaging device further comprising an external airtight cover including a planar movable airtight portion that seals between the upper part of the member and the periphery of the opening. The timer according to claim 5, further comprising a sealing flange connected to the outer surface of the coupling portion of the external airtight cover at the upper end of the mounting member and sliding up and down while maintaining airtightness with respect to the outer surface of the coupling portion when the mount member moves up and down. Labs culture cell imaging device.

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