KR20160123047A

KR20160123047A - Apparatus for counting of cells

Info

Publication number: KR20160123047A
Application number: KR1020150053064A
Authority: KR
Inventors: 임태규
Original assignee: 주식회사 지비에스코리아; 임태규
Priority date: 2015-04-15
Filing date: 2015-04-15
Publication date: 2016-10-25

Abstract

Disclosed is a cell counting device. The cell counting device according to an embodiment of the present invention comprises: a film-type cell cartridge having a chamber for feeding cells to be observed; a light source unit disposed on the cell cartridge, and irradiating light toward the cell cartridge; an image sensor disposed at the bottom of the cell cartridge, sensing light passing through the cell cartridge, and converting the light into images; and an image analyzing unit for counting the number of cells included in the images by analyzing the images. The overall area of the chamber is scanned through light irradiated by the light source unit so as to obtain images corresponding to the overall area of the chamber.

Description

[0001] APPARATUS FOR COUNTING OF CELLS [0002]

The present invention relates to a cell counting apparatus, and more particularly, to a cell counting apparatus capable of improving the accuracy of cell counting compared to existing apparatuses by scanning the entire region of a chamber (cell cartridge) accommodating cells to be observed Counting device.

Cell kinetic analysis refers to the analysis of temporal changes in the shape, growth rate, etc. of cell populations. For example, cell activity analysis is essential to confirm the effects of anti-cancer drugs and other factors in the cancer cell field, and to determine whether hepatocytes during culturing exhibit normal proliferative capacity in hepatocyte analysis.

In cell activity analysis, cell density measurement is the main indicator, and cell density measurement is based on the measurement of cell number, so-called 'cell counting'.

The cell counting device refers to a device that performs cell counting automatically by optical sensing or the like, out of the conventional manual counting method using microscopic observation.

As a representative cell counting apparatus, there is a cell counting apparatus including a flow cytometer for measuring excitation light to cells passing through a capillary column and detecting fluorescence generated therefrom, And image cytometry in which excitation light is irradiated to a certain region and a fluorescence image emitted therefrom is detected by a CCD.

However, there is a limit to the fact that flow cytometry is not an image sensing method, and image cytometry estimates the total number of cells through image of a part of the cell chamber, However, there is also the disadvantage that the devices are too expensive to be sold widely.

FIG. 12 schematically shows a conventional cell counting apparatus 10 in which a CMOS sensor is adopted, unlike the flow cytometry or image cytometry described above.

The conventional cell counting apparatus 10 includes a light source unit 14 having a light source 14a, a CMOS image sensor 15, and an objective lens 19. The light source unit 14 irradiates the light L toward the cell cartridge 12 accommodating the observation target cells and the light that has passed through the cell cartridge 12 is guided by the objective lens 19 to the CMOS image sensor 15 As shown in FIG.

Although this type of cell counting apparatus 10 is advantageous in that the image of the target cells obtained through the CMOS image sensor 15 is clear because the optical focusing is performed by the objective lens 19, Since the method of estimating the number and density of all the cells is applied through the cell counting for the objective lens 19, the reliability of the accuracy is not high. In addition, the objective lens 19, together with the CMOS image sensor 15, There is a limit in reducing the manufacturing cost.

In the case of the cell counting apparatus 10 described above, the reason why most of the light L incident on the cell cartridge 12 is obliquely incident on the cell cartridge 12 is that the image obtained by the CMOS image sensor 15 The shadows of some cells may also appear on the cell, resulting in a disadvantage that the accuracy of the cell counting is degraded.

The main object of the present invention is to provide a cell counting apparatus capable of not only increasing the accuracy of cell counting but also reducing manufacturing cost.

In order to achieve the above object, the present invention provides a cell cartridge comprising: a cell-shaped cell cartridge having a chamber into which cells to be observed are injected; A light source unit disposed above the cell cartridge and irradiating light toward the cell cartridge; An image sensor disposed under the cell cartridge and detecting light passing through the cell cartridge and converting the light into an image; And an image analysis unit for counting the number of cells included in the image through analysis of the image, wherein the image is obtained through light emitted by the light source unit such that an image corresponding to the total area of the chamber is obtained And the entire area of the chamber is scanned.

The cell counting device may further include a cell cartridge driving unit for moving the cell cartridge in a horizontal direction for scanning the entire area of the chamber.

The cell cartridge driving unit may further include: a cell cartridge supporting member on which the cell cartridge is mounted; And a cell cartridge driving device for horizontally driving the cell cartridge support.

The cell counting device comprises: an image sensor support for supporting the image sensor; And an image sensor driving device for horizontally driving the image sensor support so that an image corresponding to the entire area of the chamber can be obtained.

The light source unit may irradiate parallel light toward the cell cartridge. An air layer may be present as an optical element between the cell cartridge and the image sensor so that light passing through the cell cartridge can be incident on the image sensor in a parallel light form.

The light source unit includes: a light source for providing the light; And an optical component for converting the light provided by the light source into a parallel light form. The optical component may be an optical lens.

The distance between the cell cartridge and the image sensor is preferably within 2 mm.

The cell counting apparatus may further include a main casing in which a cell cartridge slot for receiving the cell cartridge is formed, and the light source unit and the image sensor may be spaced apart from each other with the cell cartridge interposed therebetween .

According to the cell counting apparatus of the present invention, since the optical sensing is performed on the entire area of the chamber in the cell cartridge, the accuracy of the cell counting can be greatly improved as compared with the conventional cell counting apparatuses.

In addition, since the light irradiated toward the cell cartridge by the light source unit is provided in a parallel light form, a cell image having a resolution sufficient for cell counting can be obtained, and the number of light sources required for optical sensing can be minimized.

In addition, since the parallel light provided by the light source unit is vertically incident on the cell cartridge, shadows of some of the cells appearing in the conventional cell counting device are not displayed on the image obtained by the image sensor, The accuracy of the counting can be further improved.

Since the objective lens for focusing the light on the image sensor is not used, the manufacturing cost of the cell counting device can be reduced compared with the conventional case.

1 is a perspective view schematically illustrating a cell counting apparatus according to an embodiment of the present invention.
Figure 2 is a schematic cross-section of the cell counting device of Figure 1;
3 is a schematic plan view of a cell catridge provided in the cell counting apparatus of FIG.
4 is a schematic cross-sectional view along the line I-I of the cell cartridge of Fig. 3;
5 is a cross-sectional view illustrating a cell counting process performed by the cell counting apparatus of FIG.
6 is a plan view of the cell cartridge of Fig. 5 in which cell counting is being performed.
Figures 7 to 10 are schematic diagrams for explaining alternative embodiments of the cell counting apparatus according to the present invention.
11 is an enlarged view showing an example of an image of animal cells obtained by the cell counting device of FIG.
12 is a view schematically showing an example of a conventional cell counting apparatus employing a CMOS image sensor.

1 is a schematic perspective view of a cell counting apparatus according to an embodiment of the present invention, FIG. 2 is a schematic cross-sectional view of the cell counting apparatus of FIG. 1, and FIG. 3 is a cross- FIG. 4 is a schematic cross-sectional view of the cell cartridge of FIG. 3. FIG.

1 to 4, a cell counting apparatus 100 according to an embodiment of the present invention includes a main body casing 110, a cell cartridge 120, a cell cartridge driving unit 130, a light source unit 140, A sensor 150, an image sensor support 160, an image analysis unit 170, and an image display unit 180.

The main body casing 110 provides a space for optical sensing of cells in the cell cartridge 120. The main body casing 110 includes a cell cartridge driving unit 130, a light source unit 140, And an image sensor 150 are mounted and supported. Although the main body casing 110 has a substantially rectangular parallelepiped shape in this embodiment, it is needless to say that the shape of the main body casing 110 can be variously changed.

A cell cartridge slot 111 is formed on one side of the main casing 110. The cell cartridge 120 can be moved in and out of the main body casing 110 through the cell cartridge slot 111. [

The cell cartridge 120 is a part for accommodating and transporting observation target cells to be counted in a cell. As shown in FIGS. 3 and 4, the cell cartridge 120 can be made of a transparent film having a chamber 121 in which cells to be observed are injected and accommodated. Two openings 123 and 124 communicating with the chamber 121 may be formed in the cell cartridge 120. The cells to be observed can be injected into the chamber 121 through one of the two openings 123 and 124 while the other opening serves as an air outlet through which the air in the chamber 121 is exhausted.

The cell cartridge driving unit 130 serves to support and move the cell cartridge 120 into which the observation target cells are injected. The cell cartridge driving unit 130 includes a cell cartridge support 131 and a cell cartridge driving device 132.

The cell cartridge support 131 supports the cell cartridge 120 and is mounted in the main casing 110 movably in the horizontal direction. For example, the cell cartridge support 131 may be movably provided along the X direction or may be provided movably along the X and Y directions. Thus, the cell cartridge support 131 can move the cell cartridge 120 supported thereon in the X direction through movement in the horizontal direction (X direction in the drawing). The cell cartridge 120 supported by the cell cartridge support 131 can be moved in and out of the main body casing 110 through the cell cartridge slot 111 as well as moved in the main casing 110 in the horizontal direction do.

The cell cartridge driving device 132 drives the cell cartridge support 131 and is provided as a linear motor in the present embodiment. However, the cell cartridge driving device 132 may be a linear motor, Of course, can be applied.

The light source unit 140 is disposed on the cell cartridge 120 and irradiates light toward the chamber 121 of the cell cartridge 120. As shown in FIG. 2, the light source unit 140 may be mounted on the upper portion of the main body casing 110.

The light source unit 140 includes a light source 141 for providing light and an optical component 142 for converting the light into a parallel light form. As an example, the light source 141 may be provided with an LED, and the optical component 142 may be provided with an optical lens. By providing such an optical component 142, the optical unit 140 irradiates parallel light to the cell cartridge 120.

The image sensor 150 is disposed below the cell cartridge 120 and at a position facing the light source unit 140 described above. The light emitted from the light source unit 140 passes through the chamber 121 of the cell cartridge 120 and is then sensed by the image sensor 150. Through the light sensing, It is possible to generate an image of cells.

In this embodiment, the image sensor 150 is provided with a complementary metal-oxide semiconductor (CMOS) sensor. However, the present invention is not limited thereto, and other types of image sensors capable of generating images of cells to be observed through optical sensing may be applied.

The distance d between the image sensor 150 and the above-described cell cartridge 120 (see FIG. 5) is preferably as close as possible. If the distance d between the image sensor 150 and the cell cartridge 120 is too long, the resolution of the image sensed by the image sensor 150 may be too low to reduce the accuracy of the cell counting. Therefore, in this embodiment, the distance d between the image sensor 150 and the cell cartridge 120 is set to 2 mm or less. However, for convenience of illustration, the distance d is somewhat exaggerated in the drawing.

The image sensor support 160 is a part for supporting the image sensor 150 and can be mounted on the lower side of the main body casing 110 in the main body casing 110 as shown in Fig. In an alternative embodiment, the image sensor support 160 may be omitted by replacing the lower surface of the main body casing 110 with the image sensor support 160.

An image sensor drive device (e.g., a linear motor) may be connected to the image sensor support 160 according to an embodiment. In this case, since the image sensor support 160 is driven in the horizontal direction, the image sensor 150 mounted on the image sensor support 160 can be moved in the same direction. In this case, for example, the image sensor support 160 may be movably provided along the X direction and the Y direction by the image sensor driving device, or may be provided movably along the X direction and the Y direction .

The image analysis unit 170 analyzes the image generated by the image sensor 160 to count the number of cells included in the image. In this image analysis process, the image analysis unit can use a known image analysis program (algorithm). Here, since the image analysis program itself is not an essential feature of the present invention, a detailed description thereof will be omitted.

The image analysis unit 170 may distinguish between living and dead cells among the cells accommodated in the cell cartridge 120 through the image analysis described above. For example, when animal cells immersed in a biochemical staining solution such as a trypan blue solution are housed in the cell cartridge 120, dead cells in which cell membranes are destroyed are stained blue On the other hand, living cells without destruction of cell membranes are not stained so that they are colored differently. In this case, the image analysis unit 170 analyzes the color of the cells to determine the number of living cells and the number of dead cells You can count.

For example, the image analysis unit 170 may be provided with a microprocessor equipped with the above-described image analysis program, and the microprocessor may be provided in the form of a printed circuit board (PCB), for example.

The image display unit 180 displays an image of cells generated by the image sensor 150. The cell counting result performed by the image analysis unit 170 may also be displayed as an image. The image display unit 180 may be implemented by various displays such as an LCD and an OLED, and may be provided as a PC monitor (personal computer monitor), for example.

2, the image analysis unit 170 and the image display unit 180 are shown disposed outside the body casing 110, but in an alternative embodiment, the image analysis unit 170 may include a body casing (not shown) 110, while the image display unit 180 may be mounted in one area of the outer surface of the main body casing 110.

5 is a cross-sectional view for explaining a cell counting process performed by the cell counting apparatus 100, and FIG. 6 is a plan view of the cell cartridge of FIG. 5 in which cell counting is performed. The cell counting process will be described with reference to the drawings.

First, a cell cartridge 120 containing cells to be observed in a chamber 121 is inserted into the main casing 110 (see FIG. 2) while being mounted on the cell cartridge support 131 (FIG. 2).

Next, the light source unit 140 is used to illuminate the cell cartridge 120 with the light L for image sensing of the cells to be observed. The light source unit 140 is provided with the optical component 142 that converts the light generated by the light source 141 into the parallel light so that the light L emitted from the light source unit 140 is in the form of a parallel light . Since the air layer exists between the cell cartridge 120 and the image sensor 150 and no other optical element exists, the parallel light passing through the chamber 121 of the cell cartridge 120 is still parallel light And may be received by the image sensor 150 in the form of an image.

The cell cartridge 120 is moved in the horizontal direction (X direction in this embodiment) through the cell cartridge drive unit 130 (see FIG. 2) while irradiating the parallel light through the light source unit 140. Optical scanning of the entire region of the chamber 121 in the cell cartridge 120 is performed through the horizontal movement of the cell cartridge 120 so that the cells extending over the entire region of the chamber 121 by the image sensor 150 May be generated.

Next, the image analysis unit 170 analyzes the image obtained by the image sensor 150 and counts the number of cells included in the image. Since the image obtained by the image sensor 150 is obtained by scanning the entire area of the chamber 121, the image analysis unit 170 can accurately count the total number of cells existing in the chamber 121. [ The image analysis unit 170 may also calculate the cell density based on the number of counted cells. The number of cells or the cell density calculated by the image analysis unit 170 may be provided to the user by being displayed through the image display unit 180.

Figures 7 to 10 are schematic diagrams for explaining alternative embodiments of the cell counting apparatus according to the present invention.

7, the light source unit 140 includes a plurality of light sources 141 and a plurality of optical components 142, and is arranged in parallel to cover the entire area of the chamber 121 toward the cell cartridge 120. In this embodiment, Light. Therefore, the cell cartridge 120 does not need to be moved horizontally for optical sensing of the entire area of the chamber 121. [ Instead, the image sensor 150 disposed under the cell cartridge 120 is horizontally moved along the longitudinal direction (X direction) of the cell cartridge 10.

8, the light source unit 140 includes a plurality of light sources 141 and a plurality of optical components 142, and is arranged in parallel to cover the entire area of the chamber 121 toward the cell cartridge 120. In other words, Light. A number of image sensors 150 are provided on the lower side of the cell cartridge 120 to cover the entire area of the chamber 121. Therefore, in the case of the embodiment of FIG. 8, horizontal movement of the image sensor 150 as well as horizontal movement of the cell cartridge 120 is not required for optical sensing of the entire area of the chamber 121.

9, a plurality of image sensors 150 arranged in a line are provided, and the light source unit 140 provides parallel light covering the entire area thereof toward the plurality of image sensors 150 . During optical scanning, the cell cartridge 120 is horizontally moved along its width direction (Y direction) so that light sensing with respect to the entire area of the plurality of chambers 121 can be achieved.

10, the cell cartridge 120 includes a plurality of chambers 121. The light source unit 140 covers the entire area of the plurality of chambers 121 toward the cell cartridge 120 Parallel light. A plurality of image sensors 150 arranged in a line are horizontally moved along the width direction (Y direction) of the cell cartridge 120 so that optical sensing with respect to the entire area of the plurality of chambers 121 can be achieved. At this time, the cell cartridge 120 is not moved during optical sensing.

8, 9, and 10, a plurality of image sensors 150 are provided. Instead of having a plurality of image sensors 150, a plurality of image sensors 150 may be provided in the cell cartridges 120, A line image sensor of a size that can cover the entire length of the line image sensor.

11 is an enlarged view of an example of an image of animal cells obtained by the above-described cell counting apparatus 100, wherein the animal cells are mixed with a trypan blue solution, ). As shown in FIG. 11, the cells (C1) that are not destroyed by cell membranes are whitened because they are not stained with trypan blue, whereas the cells (C2) that are destroyed by cell membranes are seen as blue by staining with trypan blue. The cell counting apparatus 100 can count the total number of cells C1 and C2 as well as distinguish the living ones C1 from the dead ones C2 by color analysis of the cell image, Cell counting may also be performed.

According to the cell counting apparatus of the present invention, since the light sensing is performed on the entire area of the chamber in the cell cartridge, the accuracy of the cell counting can be greatly improved as compared with the conventional cell counting apparatuses.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined in the appended claims. It can be understood that

100: Cell counting device
110: main housing
111: Cell cartridge slot
120: cell cartridge
130: cell cartridge driving unit
131: cell cartridge support
132: cell cartridge driving device
140: Light source unit
141: Light source
142: Optical component
150: Image sensor
160: Image sensor support
170: image analysis unit
180: Image processing unit

Claims

A cell-shaped cell cartridge having a chamber into which cells to be observed are injected;
A light source unit disposed above the cell cartridge and irradiating light toward the cell cartridge;
An image sensor disposed under the cell cartridge and detecting light passing through the cell cartridge and converting the light into an image; And
And an image analysis unit for counting the number of cells included in the image through analysis of the image,
Wherein the entire area of the chamber is scanned through light illuminated by the light source unit such that an image corresponding to the total area of the chamber can be obtained.
Cell counting device.

The method according to claim 1,
Further comprising a cell cartridge driving unit for moving the cell cartridge in a horizontal direction for scanning the entire area of the chamber,
Cell counting device.

3. The method of claim 2,
The cell cartridge driving unit includes:
A cell cartridge support on which the cell cartridge is mounted; And
And a cell cartridge driving device for horizontally driving the cell cartridge support,
Cell counting device.

The method according to claim 1,
An image sensor support for supporting the image sensor; And
Further comprising: an image sensor driving device for horizontally driving the image sensor support so that an image corresponding to an entire area of the chamber can be obtained;
Cell counting device.

The method according to claim 1,
And the light source unit irradiates the parallel light type light toward the cell cartridge,
Cell counting device.

6. The method of claim 5,
Wherein only an air layer exists as an optical element between the cell cartridge and the image sensor so that light passing through the cell cartridge can be incident on the image sensor in a parallel light form.
Cell counting device.

6. The method of claim 5,
The light source unit includes:
A light source for providing the light; And
And an optical component that converts the light provided by the light source into a parallel light form.
Cell counting device.

8. The method of claim 7,
Wherein the optical component is an optical lens,
Cell counting device.

The method according to claim 1,
Wherein the distance between the cell cartridge and the image sensor is within 2 mm,
Cell counting device.

The method according to claim 1,
Wherein the light source unit and the image sensor are spaced apart from each other with the cell cartridge interposed therebetween, wherein the light source unit and the image sensor are spaced apart from each other,
Cell counting device.