WO2019196043A1 - Cell culture device and method for detecting cell using same - Google Patents
Cell culture device and method for detecting cell using same Download PDFInfo
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- WO2019196043A1 WO2019196043A1 PCT/CN2018/082739 CN2018082739W WO2019196043A1 WO 2019196043 A1 WO2019196043 A1 WO 2019196043A1 CN 2018082739 W CN2018082739 W CN 2018082739W WO 2019196043 A1 WO2019196043 A1 WO 2019196043A1
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- 238000004113 cell culture Methods 0.000 title claims abstract description 50
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- 238000000799 fluorescence microscopy Methods 0.000 description 2
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- the invention belongs to the technical field of microscopic imaging, and in particular relates to a cell culture device and a method for detecting the same.
- the excitation light path is to irradiate the sample from the side with a thin layer of light, and then detect the fluorescence from the upper or lower portion of the sample, and the excitation light path and the detection light path are perpendicular to each other.
- the traditional inverted microscope used to detect living cells using a laser confocal Petri dish with a 0.17mm slide at the bottom and a plastic material on the side, about 1mm thick, beyond the depth of detection of the high magnification objective, in the excitation path and detection
- the excitation source objective lens cannot be close to the sample from the side, so the confocal cell culture dish cannot be used for cell sample culture of the light microscopy technique.
- two glass flakes are usually vertically adhered together, and samples such as cells are usually dispersed in a gel and placed on a horizontal slide in the form of a solid, wherein the objective lens of the excitation light source is attached.
- the objective lens of the detection source is attached to the horizontal slide.
- this method has significant disadvantages: 1. It is easy to damage; 2. The sample is easy to leak; 3. The slide is difficult to maintain the vertical state, making the angle of the excitation light path and the detection light path difficult to control.
- the technical problem to be solved by the present invention is to provide a cell culture device and a method for detecting the same, which aim to solve the problem that the cell culture device for the light film imaging technology in the prior art is easily damaged, the sample is easily leaked, and the excitation light path and the detection are performed. It is difficult to keep the light path vertical.
- the present invention is achieved by a cell culture device for a light film imaging technology, the cell culture device is a cubic type, including four sides and a bottom surface;
- a first window is defined in the bottom surface, and the first window is provided with a first sheet matching the shape of the first window to transmit light, and the first sheet is parallel to the bottom surface;
- a second window is disposed on at least one of the sides, and the second window is provided with a second sheet matching the shape of the second window to transmit light, the second sheet being parallel to the side thereof .
- the cell culture device further includes a top cover for snapping over the top surface of the cell culture device.
- the shape of the first window is a circle or a square.
- the shape of the second window is a circle or a square.
- the material of the first sheet is transparent glass or transparent plastic.
- the material of the second sheet is transparent glass or transparent plastic.
- first sheet and the second sheet each have a thickness of 0.17 mm.
- the present invention also provides a method for detecting cells using the cell culture device described above, the method comprising the steps of:
- a test objective is attached to the outer side of the first sheet of the cell culture device for detecting the state of the cell sample.
- the present invention has the beneficial effects that the cell culture device of the present invention is a cubic type, including four sides and a bottom surface, and the first window is opened on the bottom surface, and the first window is provided with the first window.
- a shape-matching first sheet that transmits light the first sheet being parallel to the bottom surface; at least one side having a second window disposed therein, the second window being provided with a second sheet matching the shape of the second window for transmitting light
- the second sheet is parallel to the side on which it is located.
- the cell culture device of the present invention adopts a cubic structure instead of a vertical adhesion of the two slides, so that the cell culture device is not easily damaged, and the sample is not easily leaked, and at the same time, since the first sheet is parallel to the bottom surface.
- the second sheet is parallel to the side on which it is located, and the bottom surface of the cube is perpendicular to the side surface. Therefore, if the excitation objective lens is closely attached to the outer side of the second sheet, and the detection objective lens is in close contact with the outer side of the first sheet, the excitation light path is perpendicular to the detection light path. Relationship, thereby improving the utility of cell culture devices for light film imaging techniques.
- FIG. 1 is a schematic structural view of a cell culture apparatus according to an embodiment of the present invention.
- Figure 2 is a schematic view showing the structure of another angle of Figure 1;
- Figure 3 is a schematic view showing the structure of still another angle of Figure 1.
- a cell culture device As shown in FIG. 1 to FIG. 3, a cell culture device according to an embodiment of the present invention is mainly applied to a light film imaging technology, and the cell culture device is a cubic type, including a bottom surface 1 and four side surfaces 2 .
- a first window is defined in the bottom surface 1.
- the first window is provided with a first sheet 101 matching the shape of the first window to transmit light.
- the first sheet 101 is parallel to the bottom surface 1.
- a second window is defined in the at least one side 2, and the second window is provided with a second sheet 201 matching the shape of the second window to transmit light, and the second sheet 201 is parallel to the side 2 on which it is located.
- the second window may be opened on one side 2 or the second window may be opened on the four sides 2, which may be selected according to specific needs, and is not limited herein.
- the above cell culture device further includes a top cover 3 for capping on the top surface of the cell culture device to keep the cell culture device clean.
- the shapes of the first window and the second window can be designed as a circle, a square, a triangle, or other various shapes, which are not limited herein.
- the material of the first sheet 101 and the second sheet 102 may be transparent glass or transparent plastic.
- the thickness of the first sheet 101 and the second sheet 102 can both be set to 0.17 mm.
- the method for detecting cells by using the cell culture device comprises the following steps:
- the detection objective is attached to the outer side of the first sheet of the cell culture device, and the detection objective is used to detect the state of the cell sample.
- the cell culture device provided by the embodiment and the method for detecting the same, wherein the cell culture device adopts a cubic structure instead of the vertical adhesion of the two slides, so that the cell culture device is not easily damaged, and the sample is not easily leaked.
- the first sheet is parallel to the bottom surface
- the second sheet is parallel to the side thereof, and the bottom surface of the cube is perpendicular to the side surface. Therefore, if the excitation objective lens is closely attached to the outer side of the second sheet, the detecting objective lens is closely attached to the outer side of the first sheet.
- the excitation light path is perpendicular to the detection light path, thereby improving the utility of the cell culture device for the light film imaging technique.
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Abstract
A cell culture device and a method for detecting a cell using same. The cell culture device has a cubic shape, and comprises a bottom face (1) and four side faces (2); a first window is provided on the bottom face (1), and the first window is internally provided with a first sheet (101) which has a shape matching the shape of the first window and allows light to be transmitted, the first sheet (101) being parallel to the bottom face (1); a second window is provided on at least one side face, and the second window is internally provided with a second sheet (201) which has a shape matching the shape of the second window and allows light to be transmitted, the second sheet (201) being parallel to the side face (2) where the second sheet is located. The cell culture device has a cubic structure rather than a structure where two glass slides are adhered perpendicularly to each other, so that the cell culture device is not easily damaged, and a sample does not easily leak, also perpendicularity between an excitation light path and a detection light path can be ensured, thereby improving the practicability of the cell culture device for light sheet imaging technology.
Description
本发明属于显微成像技术领域，尤其涉及一种细胞培养装置及应用其检测细胞的方法。The invention belongs to the technical field of microscopic imaging, and in particular relates to a cell culture device and a method for detecting the same.
随着科技的进步，新一代的光学显微成像技术得到了巨大的发展，如光片荧光显微技术。在光片荧光显微技术中，激发光路是利用薄层光从侧面照射样本，然后从样本的上部或下部检测荧光，激发光路与检测光路相互垂直。传统的倒置显微镜检测活体细胞时，使用的是激光共聚焦培养皿，其底部为0.17mm的玻片，侧面为塑料材质，厚度约为1mm，超出了高倍物镜的探测深度，在激发光路与检测光路相互垂直的情况下，激发光源物镜无法从侧面贴近样品，因此共聚焦细胞培养皿不能用于光片显微技术的细胞样品培养。目前制备光片成像的细胞样品时，通常是用两个玻璃片垂直粘连在一起，细胞等样品通常是分散在凝胶中，以固体的形式放置在水平的玻片上，其中激发光源的物镜贴在垂直方向的玻片上，而探测光源的物镜贴在水平方向的玻片上。不过这种方法存在显著的缺点：1.容易损坏；2.样品容易泄露；3.玻片难以保持垂直状态，使激发光路与检测光路的角度很难控制。With the advancement of technology, a new generation of optical microscopy imaging technology has been greatly developed, such as light sheet fluorescence microscopy technology. In the light sheet fluorescence microscopy technique, the excitation light path is to irradiate the sample from the side with a thin layer of light, and then detect the fluorescence from the upper or lower portion of the sample, and the excitation light path and the detection light path are perpendicular to each other. The traditional inverted microscope used to detect living cells, using a laser confocal Petri dish with a 0.17mm slide at the bottom and a plastic material on the side, about 1mm thick, beyond the depth of detection of the high magnification objective, in the excitation path and detection When the optical paths are perpendicular to each other, the excitation source objective lens cannot be close to the sample from the side, so the confocal cell culture dish cannot be used for cell sample culture of the light microscopy technique. At present, when preparing a cell sample for photolithography, two glass flakes are usually vertically adhered together, and samples such as cells are usually dispersed in a gel and placed on a horizontal slide in the form of a solid, wherein the objective lens of the excitation light source is attached. On the slide in the vertical direction, the objective lens of the detection source is attached to the horizontal slide. However, this method has significant disadvantages: 1. It is easy to damage; 2. The sample is easy to leak; 3. The slide is difficult to maintain the vertical state, making the angle of the excitation light path and the detection light path difficult to control.
本发明所要解决的技术问题为提供一种细胞培养装置及应用其检测细胞的方法，旨在解决现有技术中用于光片成像技术的细胞培养装置容易损坏，样品容易泄露，激发光路与检测光路难以保持垂直的问题。The technical problem to be solved by the present invention is to provide a cell culture device and a method for detecting the same, which aim to solve the problem that the cell culture device for the light film imaging technology in the prior art is easily damaged, the sample is easily leaked, and the excitation light path and the detection are performed. It is difficult to keep the light path vertical.
为解决上述技术问题，本发明是这样实现的，一种用于光片成像技术的细胞培养装置，所述细胞培养装置为立方体型，包括四个侧面及一个底面；In order to solve the above technical problem, the present invention is achieved by a cell culture device for a light film imaging technology, the cell culture device is a cubic type, including four sides and a bottom surface;
所述底面上开设有第一窗口，所述第一窗口内设置有与所述第一窗口形状匹配的可使光线透射的第一薄片，所述第一薄片与所述底面平行；a first window is defined in the bottom surface, and the first window is provided with a first sheet matching the shape of the first window to transmit light, and the first sheet is parallel to the bottom surface;
至少一个所述侧面上开设有第二窗口，所述第二窗口内设置有与所述第二窗口形状匹配的可使光线透射的第二薄片，所述第二薄片与其所在的所述侧面平行。a second window is disposed on at least one of the sides, and the second window is provided with a second sheet matching the shape of the second window to transmit light, the second sheet being parallel to the side thereof .
进一步地，所述细胞培养装置还包括顶盖，用于扣盖在所述细胞培养装置的顶面。Further, the cell culture device further includes a top cover for snapping over the top surface of the cell culture device.
进一步地，所述第一窗口的形状为圆形或方形。Further, the shape of the first window is a circle or a square.
进一步地，所述第二窗口的形状为圆形或方形。Further, the shape of the second window is a circle or a square.
进一步地，所述第一薄片的材料为透明玻璃或者透明塑料。Further, the material of the first sheet is transparent glass or transparent plastic.
进一步地，所述第二薄片的材料为透明玻璃或者透明塑料。Further, the material of the second sheet is transparent glass or transparent plastic.
进一步地，所述第一薄片和所述第二薄片的厚度均为0.17mm。Further, the first sheet and the second sheet each have a thickness of 0.17 mm.
本发明还提供了一种应用上述的细胞培养装置检测细胞的方法，所述方法包括以下步骤：The present invention also provides a method for detecting cells using the cell culture device described above, the method comprising the steps of:
a. 将细胞样品放置于所述细胞培养装置内；a. placing a cell sample in the cell culture device;
b.将激发物镜紧贴在所述细胞培养装置的第二薄片的外侧面上，所述激发物镜用于向所述细胞样品发射激发光；b. attaching an excitation objective to the outer side of the second sheet of the cell culture device, the excitation objective for emitting excitation light to the cell sample;
c.将检测物镜紧贴在所述细胞培养装置的第一薄片的外侧面上，所述检测物镜用于检测所述细胞样品的状态。c. A test objective is attached to the outer side of the first sheet of the cell culture device for detecting the state of the cell sample.
本发明与现有技术相比，有益效果在于：本发明中的细胞培养装置为立方体型，包括四个侧面及一个底面，底面上开设有第一窗口，第一窗口内设置有与第一窗口形状匹配的可使光线透射的第一薄片，第一薄片与底面平行；至少一个侧面上开设有第二窗口，第二窗口内设置有与第二窗口形状匹配的可使光线透射的第二薄片，第二薄片与其所在的侧面平行。相较于现有技术，本发明的细胞培养装置采用了立方体的结构，而不是两玻片垂直粘连的方式，使细胞培养装置不容易损坏，且样品不易泄露，同时由于第一薄片与底面平行，第二薄片与其所在的侧面平行，立方体的底面与侧面为垂直关系，因此若将激发物镜紧贴第二薄片的外侧，检测物镜紧贴第一薄片的外侧，则激发光路与检测光路为垂直关系，从而提高了用于光片成像技术的细胞培养装置的实用性。Compared with the prior art, the present invention has the beneficial effects that the cell culture device of the present invention is a cubic type, including four sides and a bottom surface, and the first window is opened on the bottom surface, and the first window is provided with the first window. a shape-matching first sheet that transmits light, the first sheet being parallel to the bottom surface; at least one side having a second window disposed therein, the second window being provided with a second sheet matching the shape of the second window for transmitting light The second sheet is parallel to the side on which it is located. Compared with the prior art, the cell culture device of the present invention adopts a cubic structure instead of a vertical adhesion of the two slides, so that the cell culture device is not easily damaged, and the sample is not easily leaked, and at the same time, since the first sheet is parallel to the bottom surface. The second sheet is parallel to the side on which it is located, and the bottom surface of the cube is perpendicular to the side surface. Therefore, if the excitation objective lens is closely attached to the outer side of the second sheet, and the detection objective lens is in close contact with the outer side of the first sheet, the excitation light path is perpendicular to the detection light path. Relationship, thereby improving the utility of cell culture devices for light film imaging techniques.
图1是本发明实施例提供的细胞培养装置的结构示意图；1 is a schematic structural view of a cell culture apparatus according to an embodiment of the present invention;
图2是图1另一角度的结构示意图；Figure 2 is a schematic view showing the structure of another angle of Figure 1;
图3是图1又一角度的结构示意图。Figure 3 is a schematic view showing the structure of still another angle of Figure 1.
为了使本发明的目的、技术方案及优点更加清楚明白，以下结合附图及实施例，对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It is understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
如图1至3所示，为本发明实施例提供的一种细胞培养装置，该细胞培养装置主要应用于光片成像技术，该细胞培养装置为立方体型，包括一个底面1及四个侧面2。As shown in FIG. 1 to FIG. 3, a cell culture device according to an embodiment of the present invention is mainly applied to a light film imaging technology, and the cell culture device is a cubic type, including a bottom surface 1 and four side surfaces 2 .
底面1上开设有第一窗口，第一窗口内设置有与第一窗口形状匹配的可使光线透射的第一薄片101，第一薄片101与底面1平行。A first window is defined in the bottom surface 1. The first window is provided with a first sheet 101 matching the shape of the first window to transmit light. The first sheet 101 is parallel to the bottom surface 1.
至少一个侧面2上开设有第二窗口，第二窗口内设置有与第二窗口形状匹配的可使光线透射的第二薄片201，第二薄片201与其所在的侧面2平行。根据不同检测的需要，可以在一个侧面2上开设第二窗口，也可以在四个侧面2上均开设第二窗口，可根据具体需要进行选择，在此不做限定。A second window is defined in the at least one side 2, and the second window is provided with a second sheet 201 matching the shape of the second window to transmit light, and the second sheet 201 is parallel to the side 2 on which it is located. Depending on the needs of the different detections, the second window may be opened on one side 2 or the second window may be opened on the four sides 2, which may be selected according to specific needs, and is not limited herein.
上述细胞培养装置还包括顶盖3，顶盖3用于扣盖在细胞培养装置的顶面上，以保持细胞培养装置的洁净。The above cell culture device further includes a top cover 3 for capping on the top surface of the cell culture device to keep the cell culture device clean.
第一窗口及第二窗口的形状均可以设计为圆形、方形、三角形或者其它各种形状，在此不做限定。The shapes of the first window and the second window can be designed as a circle, a square, a triangle, or other various shapes, which are not limited herein.
第一薄片101及第二薄片102的材料可以用透明玻璃或者透明塑料。The material of the first sheet 101 and the second sheet 102 may be transparent glass or transparent plastic.
第一薄片101和第二薄片102的厚度均可设置为0.17mm。The thickness of the first sheet 101 and the second sheet 102 can both be set to 0.17 mm.
本实施例中应用该细胞培养装置检测细胞的方法，包括以下步骤：In the embodiment, the method for detecting cells by using the cell culture device comprises the following steps:
a. 将细胞样品放置于细胞培养装置内；a. placing the cell sample in a cell culture device;
b.将激发物镜紧贴在细胞培养装置的第二薄片的外侧面上，激发物镜用于向细胞样品发射激发光；b. attaching the excitation objective lens to the outer side surface of the second sheet of the cell culture device, and the excitation objective lens is for emitting excitation light to the cell sample;
c.将检测物镜紧贴在细胞培养装置的第一薄片的外侧面上，检测物镜用于检测细胞样品的状态。c. The detection objective is attached to the outer side of the first sheet of the cell culture device, and the detection objective is used to detect the state of the cell sample.
本实施例提供的细胞培养装置及应用其检测细胞的方法，其中的细胞培养装置采用了立方体的结构，而不是两玻片垂直粘连的方式，使细胞培养装置不容易损坏，且样品不易泄露，同时由于第一薄片与底面平行，第二薄片与其所在的侧面平行，立方体的底面与侧面为垂直关系，因此若将激发物镜紧贴第二薄片的外侧，检测物镜紧贴第一薄片的外侧，则激发光路与检测光路为垂直关系，从而提高了用于光片成像技术的细胞培养装置的实用性。The cell culture device provided by the embodiment and the method for detecting the same, wherein the cell culture device adopts a cubic structure instead of the vertical adhesion of the two slides, so that the cell culture device is not easily damaged, and the sample is not easily leaked. At the same time, since the first sheet is parallel to the bottom surface, the second sheet is parallel to the side thereof, and the bottom surface of the cube is perpendicular to the side surface. Therefore, if the excitation objective lens is closely attached to the outer side of the second sheet, the detecting objective lens is closely attached to the outer side of the first sheet. The excitation light path is perpendicular to the detection light path, thereby improving the utility of the cell culture device for the light film imaging technique.
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。The above is only the preferred embodiment of the present invention, and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection of the present invention. Within the scope.
Claims (8)
- 一种细胞培养装置，其特征在于，所述细胞培养装置为立方体型，包括四个侧面及一个底面；
所述底面上开设有第一窗口，所述第一窗口内设置有与所述第一窗口形状匹配的可使光线透射的第一薄片，所述第一薄片与所述底面平行；
至少一个所述侧面上开设有第二窗口，所述第二窗口内设置有与所述第二窗口形状匹配的可使光线透射的第二薄片，所述第二薄片与其所在的所述侧面平行。A cell culture device, characterized in that the cell culture device is of a cubic type, comprising four sides and a bottom surface;
a first window is defined in the bottom surface, and the first window is provided with a first sheet matching the shape of the first window to transmit light, and the first sheet is parallel to the bottom surface;
a second window is disposed on at least one of the sides, and the second window is provided with a second sheet matching the shape of the second window to transmit light, the second sheet being parallel to the side thereof . - 如权利要求1所述的细胞培养装置，其特征在于，所述细胞培养装置还包括顶盖，用于扣盖在所述细胞培养装置的顶面。The cell culture apparatus according to claim 1, wherein said cell culture apparatus further comprises a top cover for clasping a top surface of said cell culture apparatus.
- 如权利要求1所述的细胞培养装置，其特征在于，所述第一窗口的形状为圆形或方形。The cell culture apparatus according to claim 1, wherein said first window has a circular shape or a square shape.
- 如权利要求1所述的细胞培养装置，其特征在于，所述第二窗口的形状为圆形或方形。The cell culture apparatus according to claim 1, wherein said second window has a circular or square shape.
- 如权利要求1所述的细胞培养装置，其特征在于，所述第一薄片的材料为透明玻璃或者透明塑料。The cell culture apparatus according to claim 1, wherein the material of the first sheet is transparent glass or transparent plastic.
- 如权利要求1所述的细胞培养装置，其特征在于，所述第二薄片的材料为透明玻璃或者透明塑料。The cell culture apparatus according to claim 1, wherein the material of the second sheet is transparent glass or transparent plastic.
- 如权利要求1所述的细胞培养装置，其特征在于，所述第一薄片和所述第二薄片的厚度均为0.17mm。The cell culture apparatus according to claim 1, wherein each of said first sheet and said second sheet has a thickness of 0.17 mm.
- 一种应用如权利要求1至7任意一项所述的细胞培养装置检测细胞的方法，其特征在于，所述方法包括以下步骤：
a. 将细胞样品放置于所述细胞培养装置内；
b.将激发物镜紧贴在所述细胞培养装置的第二薄片的外侧面上，所述激发物镜用于向所述细胞样品发射激发光；
c.将检测物镜紧贴在所述细胞培养装置的第一薄片的外侧面上，所述检测物镜用于检测所述细胞样品的状态。A method for detecting cells by using the cell culture apparatus according to any one of claims 1 to 7, characterized in that the method comprises the following steps:
a. placing a cell sample in the cell culture device;
b. attaching an excitation objective to the outer side of the second sheet of the cell culture device, the excitation objective for emitting excitation light to the cell sample;
c. A test objective is attached to the outer side of the first sheet of the cell culture device for detecting the state of the cell sample.
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| WO2017094451A1 (en) * | 2015-12-04 | 2017-06-08 | 公立大学法人大阪府立大学 | Cell culture vessel and sample cell for observation use |
| CN108485967A (en) * | 2018-04-12 | 2018-09-04 | 深圳大学 | A kind of cell culture apparatus and the method using its detection cell |
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| US4321330A (en) * | 1980-04-04 | 1982-03-23 | Baker Fraser L | Tissue culture device |
| CN101528912A (en) * | 2005-09-26 | 2009-09-09 | 快速微型生物***公司 | Cassette containing growth medium |
| CN202530087U (en) * | 2012-05-06 | 2012-11-14 | 李仁哲 | Sampling device for cell culture |
| WO2017094451A1 (en) * | 2015-12-04 | 2017-06-08 | 公立大学法人大阪府立大学 | Cell culture vessel and sample cell for observation use |
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