KR100813915B1 - Cell culture detection apparatus - Google Patents

Cell culture detection apparatus Download PDF

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KR100813915B1
KR100813915B1 KR1020060106142A KR20060106142A KR100813915B1 KR 100813915 B1 KR100813915 B1 KR 100813915B1 KR 1020060106142 A KR1020060106142 A KR 1020060106142A KR 20060106142 A KR20060106142 A KR 20060106142A KR 100813915 B1 KR100813915 B1 KR 100813915B1
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incubator
light
cell culture
cell
image
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홍성민
박순섭
곽연화
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전자부품연구원
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    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/06—Means for regulation, monitoring, measurement or control, e.g. flow regulation of illumination
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    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C12M41/14—Incubators; Climatic chambers

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Abstract

An apparatus for observing cell culturing is provided to observe the cells under culturing easily without opening an incubator and acquire an image of transparent cells with a CCD camera or a cheap microscope easily. An apparatus for observing cell culturing comprises: a light(310) located as the side of an incubator(200) for cell culturing; a diffused reflection plate(320) which is located at the upper side of the incubator to make light radiated from the light be diffusedly reflected toward the incubator; a CCD camera(330) which is located at the lower side of the incubator to acquire an image of cell existing in the incubator or a camera portion having a microscope; and an image displaying portion which displays the image of the cell using a signal outputted from the camera portion, wherein the light is a luminescence light such as an optical fiber light or an LED light.

Description

세포 배양 관찰 장치{Cell culture detection apparatus}Cell culture detection apparatus

도 1은 종래 기술에 따른 세포 배양 관찰 장치 단면도,1 is a cross-sectional view of a cell culture observation apparatus according to the prior art,

도 2 및 도 3은 본 발명에 따른 세포 배양 관찰 장치 단면도.2 and 3 are cross-sectional view of the cell culture observation device according to the present invention.

<도면의 주요부분에 대한 부호의 설명><Description of the symbols for the main parts of the drawings>

200: 인큐베이터 310: 조명200: incubator 310: lighting

320: 반사판 340: 이미지 표시부320: reflector 340: image display unit

350: 자동 제어부350: automatic control unit

본 발명은 세포 배양 관찰 장치에 관한 것으로서, 더욱 상세하게는 세포의 배양과 동시에 배양되는 과정의 세포 이미지를 인큐베이터를 개방하지 않고 획득할 수 있는 장치 및 방법에 관한 것이다.The present invention relates to an apparatus for observing a cell culture, and more particularly, to an apparatus and a method for acquiring a cell image of a process of being cultured simultaneously with culturing a cell without opening the incubator.

최근 유전자 해석 기술의 진보에 수반하고, 사람을 포함하는 많은 생물에 있어서 유전자 배열이 밝혀지는 것과 동시에 해석되는 단백질 등의 유전자 산물과 질병과의 인과관계도 조금씩 해명되기 시작하고 있다. 또한, 향후 각종 단백질이나 유전자 등을 망라적 또한 통계적으로 해석하기 위하여 세포를 이용한 다양한 검사 방법이나 장치에 대한 연구가 시작되고 있다.With the recent advances in the technology of genetic analysis, the causal relationship between gene products such as proteins and diseases that are interpreted at the same time as the gene sequence is revealed in many organisms including humans is gradually being elucidated. In addition, in the future, studies on various test methods and apparatuses using cells have been started to comprehensively and statistically interpret various proteins and genes.

통상적으로 세포는 플라스틱 재질의 용기 또는 유리 접시(dish) 및 플라스크(flask)등에 파종되어, 인큐베이터(incubator)내에서 배양된다.Typically, cells are seeded in plastic containers or glass dishes and flasks, and cultured in an incubator.

이와 같은 인큐베이터의 내부는 일례로 이산화탄소 농도 5%, 온도 37℃, 습도 100%에 설정되어 세포의 배양에 적당한 환경으로 유지된다. 또한, 인큐베이터 내부에는 세포에 양분을 주는 것과 동시에 배양에 적당한 산도(pH)를 지키기 위하여 매일 혹은 수일에 한번씩 배양액의 교환이 이루어진다.The inside of such an incubator is set at, for example, a carbon dioxide concentration of 5%, a temperature of 37 ° C., and a humidity of 100% to maintain an environment suitable for culturing cells. In addition, the culture medium is exchanged every day or every few days in order to nourish the cells and to keep the pH (pH) suitable for the culture.

이와 같은 방법으로 배양중에 있는 세포의 관찰은 인큐베이터로부터 플라스틱 재질의 용기 또는 유리 접시 및 플라스크 등을 꺼내어 위상차 현미경등의 도립형 현미경을 이용한 방법이 적용된다.In this way, the observation of the cells in the culture is carried out using an inverted microscope such as a phase contrast microscope by removing a plastic container or glass dish and flask from the incubator.

그러나, 이러한 종래의 배양중에 있는 세포의 관찰 방법은 가능한 신속히 세포의 관찰을 수행하여야하며, 관찰 종류 후 세포를 인큐베이터내에 되돌려야하므로 세포의 활성이 손상되는 문제점이 있다. 이는 세포의 활성이 불안정하여 정확한 평가를 수행하는 것이 곤란한 문제를 내포하며, 세포를 인큐베이터로부터 꺼낼 경우 기타 유해 물질에 의한 오염을 배제할 수 없다.However, such a conventional method for observing cells in culture has to perform cell observation as soon as possible, and there is a problem in that cell activity is impaired since the cells must be returned to the incubator after the type of observation. This implies that the activity of the cells is unstable and it is difficult to carry out an accurate assessment, and when taken out of the incubator the contamination by other harmful substances cannot be excluded.

또한, 배양 세포는 보통 무색투명에 가까워 투과형 현미경에서는 관찰이 곤 란하다. 이로 인하여 배양액과 세포의 굴절율 차를 명암차로 변환하는 위상차 방식을 적용하여 관찰한다.In addition, cultured cells are usually colorless and transparent, which is difficult to observe under a transmission microscope. For this reason, it is observed by applying a phase difference method of converting the difference in refractive index between the culture medium and the cell to the contrast difference.

도 1은 이와 같은 위상차 현미경의 모식도도로서, 이를 참조하여 더욱 상세하게 설명하면 시료 즉, 세포가 담긴 접시(110)등을 끼운 후 상부에는 조명장치(130) 및 조명장치에 빛을 공급하는 조명 램프(140), 거울통(160), 접안 렌즈 또는 카메라(150)를 배치하고, 하측에는 대물 렌즈(120)를 배치한다. 1 is a schematic diagram of such a phase contrast microscope, which will be described in more detail with reference to the illumination that supplies light to the lighting device 130 and the lighting device at the top after inserting a sample, that is, a dish 110 containing a cell. The lamp 140, the mirror barrel 160, the eyepiece or the camera 150 are disposed, and the objective lens 120 is disposed under the lamp 140.

그러나, 이와 같은 위상차 현미경은 대물 렌즈로부터 시료를 끼우는 대향측에 설치되는 투과형이나, 공간상의 제약으로 인하여 소형 자동 배양 장치에 적용하는 것은 곤란하며, 그 가격이 매우 고가인 단점이 있다. However, such a phase contrast microscope is a transmission type provided on the opposite side to which the sample is inserted from the objective lens, but it is difficult to apply to a small automatic culture apparatus due to space limitations, and the price thereof is very expensive.

따라서, 본 발명은 난반사하는 반사판과 저가의 일반 카메라 또는 현미경을 이용하여 세포를 관찰할 수 있는 세포 배양 관찰 장치를 제공함에 그 목적이 있다.Accordingly, an object of the present invention is to provide a cell culture observing apparatus capable of observing cells using a diffusely reflecting plate and a low-cost general camera or microscope.

본 발명은 인큐베이터를 개봉하지 않고도 세포의 배양과정을 관찰 할 수 있는 세포 배양 관찰 장치를 제공함에 다른 목적이 있다.Another object of the present invention is to provide a cell culture observation apparatus capable of observing a cell culture process without opening the incubator.

본 발명은 공간상의 제약을 받지 않아 소형 자동 배양 장치 등에 다양한 분야에 적용이 용이한 세포 배양 관찰 장치를 제공함에 또 다른 목적이 있다.Another object of the present invention is to provide a cell culture observing apparatus that is easy to be applied to various fields such as a small automatic culturing apparatus without being limited in space.

본 발명에 따른 세포 배양 관찰장치는 세포 배양용 인큐베이터의 측면에 위 치하며, 상기 반사판으로 광을 조사하는 조명, 상기 인큐베이터의 상측에 위치하여 상기 조명으로부터 방사된 광을 반사시키고, 반사된 광을 인큐베이터측으로 유입되도록 하기 위한 반사판, 상기 인큐베이터의 하측에 위치하여 상기 인큐베이터의 내부에 존재하는 세포의 이미지를 획득하는 카메라부 및 상기 카메라부로부터 출력되는 신호를 이용하여 상기 세포의 이미지를 표시하는 이미지 표시부를 포함한다.Cell culture observing apparatus according to the present invention is located on the side of the cell culture incubator, the illumination to irradiate light to the reflecting plate, located on the upper side of the incubator to reflect the light emitted from the illumination, the reflected light A reflector plate for inflow to the incubator side, a camera unit positioned under the incubator to acquire an image of the cells present in the incubator, and an image display unit displaying an image of the cell using a signal output from the camera unit. It includes.

반사판은 난반사하는 재질의 것을 사용하며, 인큐베이터의 상측 및 하측은 투명한 생체 친화성 재질로 형성된 것으로서 석영 및 유리가 이에 해당한다.The reflecting plate uses a material of diffuse reflection, and the upper and lower sides of the incubator are formed of a transparent biocompatible material, and quartz and glass correspond thereto.

조명은 냉광조명으로 광섬유 조명 또는 LED 조명이 이에 해당한다.Illumination is cold light, such as fiber optic light or LED light.

이하, 첨부된 도면을 참조하여 본 발명의 바람직한 실시예를 상세히 설명하기로 한다. 이에 앞서, 본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 아니되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다. 따라서 본 명세서에 기재된 실시예와 도면에 도시된 구성은 본 발명의 가장 바람직한 일실시예에 불과할 뿐이고 본 발명의 기술적 사상을 모두 대변하는 것은 아니므로, 본 출원시점에 있어서 이를 대체할 수 있는 다양한 균등물과 변형예들이 있을 수 있음을 이해하여야 한다.Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. Prior to this, terms or words used in the specification and claims should not be construed as having a conventional or dictionary meaning, and the inventors should properly explain the concept of terms in order to best explain their own invention. Based on the principle that can be defined, it should be interpreted as meaning and concept corresponding to the technical idea of the present invention. Therefore, the embodiments described in the specification and the drawings shown in the drawings are only one of the most preferred embodiments of the present invention, and do not represent all of the technical idea of the present invention, various equivalents that can be replaced at the time of the present application It should be understood that there may be water and variations.

도 2는 본 발명에 따른 세포 배양 관찰 장치의 단면을 도시한 것으로서, 이 를 참조하여 본 발명을 더욱 상세히 설명하면 다음과 같다.2 is a cross-sectional view of a cell culture observation apparatus according to the present invention, the present invention will be described in more detail with reference to the following.

도 2에 도시된 바와 같이 조명(310)을 종래와 달리 인큐베이터(200)의 측면에 구비하여 인큐베이터(200)의 상측에 일정한 각(α)을 가지는 반사판(320)에 광을 조사하면, 반사판(320)으로부터 반사된 광이 투명한 인큐베이터(200)를 투과하여 세포의 이미지를 획득할 수 있도록 한다.As shown in FIG. 2, when the illumination 310 is provided on the side of the incubator 200, the light is irradiated onto the reflector 320 having a constant angle α on the upper side of the incubator 200. The light reflected from 320 may pass through the transparent incubator 200 to obtain an image of the cell.

이때, 반사판(320)이 가지는 각(α)은 조명으로부터 방사된 광을 인큐베이터측으로 유입하기 위한 것이다. 이러한 반사판의 각(α)은 0°~ 180°이내에서 자유롭게 설정할 수 있으며, 바람직하게는 0°초과 ~ 180°미만으로 설정한다.At this time, the angle α of the reflecting plate 320 is for introducing the light emitted from the illumination to the incubator side. The angle α of such a reflecting plate can be freely set within 0 ° to 180 °, preferably above 0 ° to less than 180 °.

반사판(320)에 광을 조사하는 조명(310)은 냉각조명을 이용한 간접 조명으로 측사 조명을 사용한다. Illumination 310 for irradiating light to the reflecting plate 320 uses side lighting as indirect illumination using the cooling light.

이때, 냉각조명은 광의 확산현상 및 발열량이 적은 고성능의 광섬유 조명, LED 조명 등이 적용될 수 있으며, 이러한 냉각조명 또한 반사판(320)으로 광을 조사하기 위하여 일정한 각(β)을 가지도록 구성하는 바람직하며, 이때, 효율적으로 반사판(320)에 광을 조사하기 위하여 그 위치를 변경할 수도 있다.At this time, the cooling light may be applied to a high-performance optical fiber illumination, LED lighting, etc., light diffusion and low heat generation, such cooling lighting is also preferably configured to have a constant angle (β) to irradiate light to the reflector 320. In this case, the position may be changed to efficiently irradiate light onto the reflecting plate 320.

본 발명의 일 실시예에서는 조명을 인큐베이터의 일측면에 구비하였으나, 반사판이 가지는 각(α)을 고려하여 조명을 인큐베이터의 타측면에 일정한 각(β)을 가지도록 구비할 수도 있다.In one embodiment of the present invention, the illumination is provided on one side of the incubator, but the illumination may be provided to have a constant angle β on the other side of the incubator in consideration of the angle α of the reflector.

카메라부(330)는 고해상도의 CCD 카메라 혹은 일반 현미경을 적용하여 세포의 이미지를 정확하게 획득할 수 있다.The camera unit 330 may accurately obtain an image of a cell by applying a high resolution CCD camera or a general microscope.

이와 같은 카메라부(330)는 도 3에 도시된 바와 같이 이미지 표시부(340)와 연결되어 있다.The camera unit 330 is connected to the image display unit 340 as shown in FIG.

이미지 표시부(340)는 카메라부(330)로부터 출력되는 신호를 이용하여 획득된 세포의 이미지를 육안으로 관찰할 수 있도록 디스플레이상에 표시한다.The image display unit 340 displays on the display to visually observe the image of the cell obtained by using the signal output from the camera unit 330.

상술한 카메라부, 이미지 표시부 및 인큐베이터는 자동제어부(350)에 연결하여 세포의 배양 및 세포의 관측을 모두 자동으로 수행할 수도 있다.The camera unit, the image display unit, and the incubator described above may be connected to the automatic controller 350 to automatically perform both cell culture and cell observation.

본 발명에 따른 세포 배양 관측장치의 구동방법은 다음과 같은 일련의 과정으로 이루어진다.The method for driving a cell culture observation device according to the present invention consists of a series of processes as follows.

우선 세포 배양중인 인큐베이터의 측면에 일정한 각을 가지고 구비된 조명으로부터 반사판에 조명이 조사된다.First, the illumination is irradiated to the reflector from the illumination provided at a constant angle on the side of the incubator in the cell culture.

조사된 빛은 반사판의 표면에서 난반사하여 반사판의 하측에 존재하는 투명한 인큐베이터 및 세포내에 입사되며, 이로 인하여 인큐베이터의 하측에 존재하는 카메라부에 세포의 이미지가 획득된다.The irradiated light is diffusely reflected at the surface of the reflecting plate to be incident on the transparent incubator and the cells existing on the lower side of the reflecting plate, whereby an image of the cell is obtained in the camera unit existing on the lower side of the incubator.

획득된 세포의 이미지는 이미지 표시부에 전달되어 디스플레이상에 시각화된다.The image of the obtained cells is transferred to an image display and visualized on the display.

따라서, 본 발명에 따른 세포 배양 관측장치는 인큐베이터를 개방하지 않고도 용이하게 배양중에 있는 세포를 관측할 수 있으며, CCD 카메라 혹은 저가의 일반 현미경을 사용하여도 투명한 세포의 이미지를 용이하게 획득할 수 있는 것이다.Therefore, the cell culture observation apparatus according to the present invention can easily observe the cells in culture without opening the incubator, and can easily obtain the image of the transparent cells even by using a CCD camera or a low-cost general microscope. will be.

또한, 구성 및 조작이 간단하여 소형화 경량화가 가능하며 이로 인하여 관측 장치의 관리와 사용이 편리하다.In addition, since the configuration and operation is simple, miniaturization and light weight are possible, and thus the management and use of the observation device is convenient.

본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적 의미로 한정해서 해석되어서는 아니되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다.The terms or words used in this specification and claims are not to be construed as being limited to their usual or dictionary meanings, and the inventors may appropriately define the concept of terms in order to best describe their invention. It should be interpreted as meaning and concept corresponding to the technical idea of the present invention based on the principle that the present invention.

따라서, 본 명세서에 기재된 실시예와 도면에 도시된 구성은 본 발명의 가장 바람직한 일실시예에 불과할 뿐이고 본 발명의 기술적 사상을 모두 대변하는 것은 아니므로, 본 출원시점에 있어서 이들을 대체할 수 있는 다양한 균등물과 변형예들이 있을 수 있음을 이해하여야 한다.Therefore, the embodiments described in the specification and the drawings shown in the drawings are only one of the most preferred embodiments of the present invention and do not represent all of the technical idea of the present invention, various modifications that can be replaced at the time of the present application It should be understood that there may be equivalents and variations.

본 발명에 따른 세포 배양 관측장치는 인큐베이터를 개방하지 않고도 용이하게 배양중에 있는 세포를 관측할 수 있으며, CCD 카메라 혹은 저가의 일반 현미경을 사용하여도 투명한 세포의 이미지를 용이하게 획득할 수 있는 효과가 있다.The cell culture observing apparatus according to the present invention can easily observe the cells in culture without opening the incubator, and it is possible to easily obtain transparent cell images even by using a CCD camera or a low-cost general microscope. have.

또한, 구성 및 조작이 간단하여 소형화 경량화가 가능하며 이로 인하여 관측 장치의 관리와 사용이 편리한 이점이 있다.In addition, since the configuration and operation is simple, miniaturization and light weight are possible, and thus there is an advantage of easy management and use of the observation device.

Claims (7)

세포 배양용 인큐베이터의 측면에 위치하는 조명; Illumination located on the side of the cell culture incubator; 상기 인큐베이터의 상측에 위치하며 상기 조명으로부터 방사된 광을 인큐베이터측으로 난반사시키는 난반사판;A diffuse reflection plate positioned above the incubator and diffusely reflecting the light emitted from the illumination to the incubator side; 상기 인큐베이터의 하측에 위치하며 상기 인큐베이터의 내부에 존재하는 세포의 이미지를 획득하는 CCD 카메라 또는 현미경을 구비한 카메라부; 및A camera unit positioned below the incubator and having a CCD camera or a microscope for acquiring an image of a cell existing inside the incubator; And 상기 카메라부로부터 출력되는 신호를 이용하여 상기 세포의 이미지를 표시하는 이미지 표시부Image display unit for displaying the image of the cell using the signal output from the camera unit 를 포함하는 세포 배양 관찰 장치.Cell culture observation device comprising a. 삭제delete 제 1 항에 있어서,The method of claim 1, 상기 인큐베이터의 상측 및 하측은 투명한 생체 친화성 재질로 형성된 세포 배양 관찰 장치.The upper and lower sides of the incubator is a cell culture observation device formed of a transparent biocompatible material. 제 3 항에 있어서,The method of claim 3, wherein 상기 투명한 생체 친화성 재질은 석영 및 유리인 세포 배양 관찰 장치.The transparent biocompatible material is a cell culture observation device is quartz and glass. 제 1 항에 있어서,The method of claim 1, 상기 조명은 냉광조명인 세포 배양 관찰 장치.The illumination is cold light illumination cell culture observation device. 제 5 항에 있어서,The method of claim 5, wherein 상기 냉광조명은 광섬유 조명 또는 LED 조명인 세포 배양 관찰 장치.The cold light is a cell culture observation device is an optical fiber light or LED light. 삭제delete
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