Cell evaluation device and cell evaluation system

Abstract

To provide a cell evaluation device that can precisely evaluate a cell or a secreted material discharged from a cell.SOLUTION: The cell evaluation device of the present invention includes: a first capturing unit for capturing a secreted material secreted by a cell; and an image acquisition unit for acquiring an image obtained by imaging observation light of the cell and light emitted from a light emission material combined with the secreted material. The image acquisition unit and the first capturing unit are arranged in that order, and the image acquisition unit acquires positional information of the cell and the secreted material.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present technology relates to a cell evaluation device and a cell evaluation system. [Background technology]

[0002] In recent years, systems for evaluating cells and secreted substances from cells have continued to evolve, and various technologies have been proposed.

[0003] For example, a technique relating to a complementary metal oxide semiconductor (CMOS) biosensor system has been proposed (see Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] US Patent Application Publication No. 2012 / 0032235 Summary of the Invention [Problem to be solved by the invention]

[0005] However, the technique proposed in Patent Document 1 may not be able to accurately and precisely evaluate cells or secretory substances released from cells.

[0006] Therefore, the present technology has been developed in consideration of such circumstances, and its main objective is to provide a cell evaluation device that can accurately evaluate cells and secretory substances released from cells with high precision. [Means for solving the problem]

[0007] The present inventors have conducted intensive research to achieve the above-mentioned objective, and as a result, have surprisingly succeeded in accurately evaluating cells and secretory substances released from cells with high precision, thereby completing the present technology.

[0008] That is, in the present technology, first, a first capture unit that captures a secretory substance secreted by a cell; and an image acquisition unit that acquires an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance, the image acquisition unit and the first capture unit are arranged in this order, The image acquisition unit acquires position information of the cells and the secretory substance.

[0009] In the cell evaluation device according to the present technology, the image acquisition unit may be a CMOS image sensor. In the cell evaluation device according to the present technology, the image acquisition unit can acquire an image obtained by capturing an image of chemiluminescence emitted by the reaction of the luminescent substance, which is an enzyme, with a substrate.

[0010] In the cell evaluation device according to the present technology, the first capture portion may include a molecule that binds non-specifically or specifically to the secretory substance. In the cell evaluation device according to the present technology, a protection part may be disposed in a region of the first capture part that does not contain the molecule that binds non-specifically or specifically to the secretory substance.

[0011] The molecule that binds non-specifically or specifically to the secreted substance may be at least one type selected from the group consisting of a first antibody, a first aptamer, and a molecularly imprinted polymer, and there may be multiple types of the first antibody.

[0012] The cell evaluation device of the present technology may further include a second capture unit that captures the cells, and in the cell evaluation device of the present technology, the image acquisition unit and the second capture unit may be arranged in this order.

[0013] In the cell evaluation device according to the present technology, the first trapping portion may be disposed around the second trapping portion.

[0014] In the cell evaluation device according to the present technology, the second capture portion may include a molecule that binds non-specifically or specifically to the cell. In the cell evaluation device according to the present technology, a protection section may be disposed in a region of the second capture section that does not contain the molecule that binds non-specifically or specifically to the cell. The molecule that binds non-specifically or specifically to the cell may be at least one type selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine, and there may be multiple types of the second antibody. In the cell evaluation device according to the present technology, a photodegradable linker may be included between the second capture unit and the image acquisition unit.

[0015] In addition, the present technology includes a first capture unit that captures a secretory substance secreted by a cell; an image acquisition unit that acquires an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance; An analysis unit that analyzes the obtained image, the image acquisition unit and the first capture unit are arranged in this order, The image acquisition unit acquires position information of the cells and the secretory substance.

[0016] In the cell evaluation device according to the present technology, the analysis unit can quantify the secretory substance based on the intensity of the light emitted from the luminescent substance bound to the secretory substance. The analysis unit can analyze the cells based on the observation light of the cells and / or brightness information of the light emitted from a luminescent substance bound to a secreted substance.

[0017] The cell evaluation device according to the present technology may further include an irradiation unit that projects the cells.

[0018] The cell evaluation device according to the present technology may further include a display unit that displays the image. The display unit may include a well image display unit based on a cell image obtained by capturing an observation light of the cell and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to the secreted substance, as well as an analysis data display unit and an analysis plot display unit based on the analysis by the analysis unit.

[0019] Furthermore, the present technology includes a first capture unit that captures a secretory substance secreted by a cell; an image acquisition unit that acquires an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance; An analysis unit that analyzes the obtained image, the image acquisition unit and the first capture unit are arranged in this order, The image acquisition unit acquires position information of the cells and the secretory substance.

[0020] Furthermore, in the present technology, a first antibody that captures a secretory substance secreted by a cell; a second antibody that captures the cells; and a CMOS image sensor for capturing an image of the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance, the CMOS image sensor, the first antibody, and the second antibody are arranged in this order; The first antibody is disposed around the second antibody, a cell evaluation device, the CMOS image sensor acquiring position information of the cell and the secretory substance, a first antibody that captures a secretory substance secreted by a cell; a second antibody that captures the cells; a CMOS image sensor for capturing an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance; An analysis unit that analyzes the acquired image, the CMOS image sensor, the first antibody, and the second antibody are arranged in this order; The first antibody is disposed around the second antibody, The CMOS image sensor acquires position information of the cells and the secretory substance. In addition, this technology: a cell capturing step of introducing a plurality of cells into a container having a first capturing part for capturing a secretory substance secreted by a cell and a second capturing part for capturing the cell disposed on a semiconductor substrate, and causing the second capturing part to capture at least one of the plurality of cells; a cell stimulating step of providing a stimulus to the plurality of cells; a secretory substance capturing step of causing the first capturing unit to capture a secretory substance secreted from at least one cell among the plurality of cells; an image acquisition step of acquiring an image obtained by capturing an observation light of at least one cell among the plurality of cells and a light emitted from a luminescent substance bound to the secretory substance; an analysis step of performing an analysis using the image obtained in the image acquisition step; Including fruit, The image includes a cell image obtained by capturing the observation light, and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to the secretory substance, Each of the cell images in the cell image image corresponds to each of the chemiluminescence images in the chemiluminescence image; The analyzing step quantifies the secretory substance for each of the cells based on the correspondence. Methods for evaluating cells are also provided. In the image acquisition step, position information of the cells and the secretory substance can be acquired. The analyzing step includes analyzing the type, size, position, shape, and At least one of the states may be analyzed. The cell evaluation method may further include a recovery step of recovering a desired cell from among the plurality of cells based on the information obtained in the analysis step. The semiconductor substrate may be a semiconductor substrate on which a photodiode is formed. The semiconductor substrate may be a component of a CMOS sensor. The image acquisition step may be performed by the CMOS sensor. The image acquisition profit In the process, the chemiluminescence emitted by the reaction of the luminescent substance, which is an enzyme, with the substrate was imaged. ChemiluminescenceAn image may be acquired. The first capture portion may include a molecule that binds non-specifically or specifically to the secreted substance. The second capture portion may include a molecule that binds non-specifically or specifically to the cell. The second capture moiety may comprise a photocleavable linker. In addition, this technology: a container including a semiconductor substrate on which a first capture unit for capturing a secretory substance secreted by a cell and a second capture unit for capturing the cell are disposed; an image acquisition unit that acquires an image obtained by capturing light emitted from a luminescent substance bound to the secretory substance captured in the first capture unit in the container and an observation light of the cell captured in the second capture unit; An analysis unit that analyzes the obtained image 、 The image includes a cell image obtained by capturing the observation light, and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to the secretory substance, Each of the cell images in the cell image image corresponds to each of the chemiluminescence images in the chemiluminescence image; The analysis unit quantifies the secretory substance for each of the cells based on the correspondence. A cell evaluation system is also provided. Effect of the Invention

[0021] According to the present technology, cells and secretory substances released from cells can be accurately evaluated with high accuracy. Note that the effects described herein are not necessarily limited, and may be any of the effects described in the present disclosure. [Brief description of the drawings]

[0022] [Figure 1] FIG. 1 is a block diagram showing an example of the configuration of a cell evaluation device according to a first embodiment to which the present technology is applied. [Diagram 2] FIG. 2 is a diagram showing an example of the configuration of a cell evaluation device according to a first embodiment to which the present technology is applied. [Diagram 3] FIG. 3 is a diagram showing an example of the configuration of a cell evaluation device according to a first embodiment to which the present technology is applied. [Figure 4] FIG. 4 is a cross-sectional view showing a configuration example of a CMOS image sensor (solid-state imaging element) provided with an antibody layer. [Diagram 5] FIG. 5 is a diagram illustrating an example of the configuration of the analysis unit. [Figure 6] FIG. 6 is a diagram illustrating an example of the configuration of the display unit. [Figure 7] FIG. 7 is a block diagram showing an example of the configuration of a cell evaluation device according to a second embodiment to which the present technology is applied. [Figure 8] FIG. 8 is a diagram showing an example of patterning of the first capture portion and the second capture portion. [Figure 9] FIG. 9 is a diagram showing an example of patterning of the first capture portion and the second capture portion. [Figure 10] FIG. 10 is a diagram showing an example of patterning of the first capture portion and the second capture portion. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] A preferred embodiment for carrying out the present technology will be described below. The embodiment described below shows an example of a typical embodiment of the present technology, and the scope of the present technology is not to be interpreted narrowly by this.

[0024] The explanation will be given in the following order. 1. Overview of the cell evaluation device related to this technology 2. First embodiment (example 1 of cell evaluation device) 2-1. Cell evaluation device 2-2. First capture section 2-3.Image acquisition section 2-4.Analysis section 2-5. Irradiation unit 2-6.Display section 3. Second embodiment (Example 2 of cell evaluation device) 3-1. Cell evaluation device 3-2. First capture portion and second capture portion, and patterning of the first capture portion and the second capture portion 3-3. Image acquisition unit, analysis unit, irradiation unit and display unit 4. Third embodiment (example of cell evaluation system) 4-1. Cell evaluation system 4-2. First capture unit, second capture unit, image acquisition unit, analysis unit, irradiation unit and display unit

[0025] <1. Overview of the cell evaluation device related to this technology> An overview of the cell evaluation device according to the present technology will be described. Methods known as ELISA and ELISPOT methods, as well as specialized systems (eg, an intracellular cytokine measurement system using flow cytometry) are already widely used to obtain substances secreted by cells.

[0026] In the ELISA method, a large number of cells are cultured in a well, and after removing the cells, only the secreted substance is collected and its secretion amount is measured. In this case, it is not possible to know what percentage of the cells secreted the secreted substance. For this reason, in order to focus on individual cells, research and development is being conducted to culture cells in smaller wells in units of one or a few cells and calculate the amount secreted by each cell.

[0027] On the other hand, the ELISPOT method involves culturing a large number of cells in a well, capturing the secreted substances of the cells with a membrane placed at the bottom of the well, and after removing the cells, reacting them with the secreted substances attached to the membrane to produce a color, which can then be counted under a microscope to calculate the number of secreting cells. However, with either method, it is difficult to know which cells secreted the substances, and it is sometimes impossible to classify individual cells when multiple cells coexist. Since then, a method has been proposed in which single cells are placed in small wells for measurement, but since individual cells are separated into wells, it is not possible to observe interactions between multiple cells coexisting. In addition, as mentioned above, the ELISPOT method requires the removal of cells before measurement, so it is impossible to take out specific cells after ELISPOT for post-analysis (such as genetic analysis).

[0028] This technology was developed in light of the above situation, and makes it possible to know which cells secreted what amount even in an environment where multiple cells coexist. In addition, it will be possible to understand the interactions occurring between different cells, such as cancer cells and immune cells, which was previously difficult to apply.

[0029] Examples of cells observed by the cell evaluation device according to the present technology include cancer cells and immune cells, and immune cells in particular include T cells, NK cells, macrophages, etc. Cells also include tissues in which a single or multiple cells gather with a certain function. Furthermore, examples of cell secretions observed by the cell evaluation device according to the present technology include IFN-γ, interleukins, chemokines, etc.

[0030] This technology can at least add quantification of secretory substances for each cell and imaging of cells to the function of ELISPOT (counting secretory cells), so it is possible to identify the cell type that released the secretory substance and its state. For example, in the co-culture of cancer cells and immune cells, it is currently difficult to know which cell released IFNγ, and it is possible to count only live cells that released IFNγ, but IFNγ is also released during apoptosis. Since this technology has at least techniques based on the ELISPOT method and the ELISA method, it is possible to reduce the amount of blood samples required. Since cell recovery is possible in principle, it is possible to reanalyze only the cells that released the secretory substance. It is possible to evaluate which cells secreted how much even in an environment where multiple cells coexist. In other words, as mentioned above, it is possible to grasp the interactions occurring between different cells such as cancer cells and immune cells, which was previously difficult.

[0031] In order to specifically explain the cell evaluation device and cell evaluation system of the present technology, a first embodiment (example 1 of a cell evaluation device), a second embodiment (example 2 of a cell evaluation device), and a third embodiment (example of a cell evaluation system) of the present technology will be described below.

[0032] 2. First embodiment (Example 1 of cell evaluation device) [2-1. Cell evaluation device] The cell evaluation device of the first embodiment (example 1 of cell evaluation device) of the present technology is a cell evaluation device that includes a first capture unit that captures a secretory substance secreted by a cell, and an image acquisition unit that acquires an image obtained by capturing observation light for the cell and light emitted from a luminescent substance bound to the secretory substance, in which the image acquisition unit and the first capture unit are arranged in this order, and the image acquisition unit acquires positional information of the cell and the secretory substance.

[0033] Here, arranging the image acquisition section and the first capture section in this order means that the image acquisition section and the first capture section may be arranged in succession, or that the image acquisition section and the first capture section may be arranged via an optional material layer (which may be a material film; the same applies below) or material section. Arranging the image acquisition section and the first capture section in succession means that the first capture section is arranged directly above the image acquisition section. Arranging the first capture section directly above the image acquisition section means, for example, that the first capture section is arranged within a distance of 150 μm, preferably within a distance of 10 μm, from the image acquisition section.

[0034] The cell evaluation device of the first embodiment according to the present technology may further include at least one of an analysis unit, an irradiation unit, and a display unit. That is, the cell evaluation device of the first embodiment according to the present technology may further include an analysis unit, an irradiation unit, a display unit, an analysis unit and an irradiation unit, an analysis unit and a display unit, an irradiation unit and a display unit, or an analysis unit, an irradiation unit, and a display unit.

[0035] A specific example of the cell evaluation device according to the first embodiment of the present technology will be described with reference to Figures 1 to 3. Figure 1 is a block diagram showing an example of the cell evaluation device according to the first embodiment of the present technology. Figures 2 and 3 are configuration diagrams showing an example of the cell evaluation device according to the first embodiment of the present technology.

[0036] First, refer to FIG. 1. The cell evaluation device 1 includes at least an upper part 101 of the image sensor including a first capture unit 101, an image sensor 1001 (e.g., a CMOS image sensor) that is an image acquisition unit, and an analysis unit 1005. The cell evaluation device 1 further includes a driver 1002, an irradiation control unit 1004, a display unit 1006, and an irradiation unit 1007. As shown in FIG. 1, the irradiation control unit 1004, the analysis unit 1005, and the display unit 1006 may be formed as one device 1003. Although not shown in FIG. 1, the upper part 101 of the image sensor may include a protection unit in addition to the first capture unit 101. The irradiation control unit 1004 can control the wavelength selectivity (color selectivity of light) of the light emitted by the irradiation unit 1007 and the exposure time of the light.

[0037] In the cell evaluation device 1, a first capture unit 101-1 (e.g., a capture unit including a first antibody for capturing a secretory substance) is placed in a desired positional relationship on an image sensor 1001. Cells can be identified by taking cell images using cell observation light or an irradiation unit 1007, and image analysis can be performed to determine cell size and cell shape. The cell type and cell state (such as whether the cell is alive or dead) can be accurately determined. In addition, the irradiation unit includes a Digital Mirror Device (DMD) or the like, and the irradiation position can also be controlled.

[0038] Then, cells are cultured on the image sensor 1001, and a secretory substance released from the cells is captured by the first capture unit 101-1 (for example, a capture unit including a first antibody for capturing the secretory substance), and then further reacted with a chemiluminescent substance or the like to generate weak light, which can be acquired by the image sensor 1001 directly below. The chemiluminescent substance is, for example, an enzyme, and chemiluminescence is emitted by the reaction of the enzyme with a substrate.

[0039] The weak light is photoelectrically converted into an electric signal by the driver 1002, and the brightness of only the sensor part where the secretory substance is captured increases, making it possible to map the position where the secretory substance was obtained on the image sensor 1001. Since the positional relationship between the first capture part 101-1 (e.g., a capture part including a first antibody for capturing the secretory substance) and the cell can be specified in advance, it is also possible to identify the cell that released the secretory substance. In addition, the analysis part 1005 can also quantify the secretory substance by utilizing the fact that the brightness changes depending on the concentration of the secretory substance. The image obtained from the image sensor 1001 is displayed on the display part 1006 (e.g., a monitor) through the analysis part 1005.

[0040] In order to increase the sensitivity, there are measures such as increasing the density of the first antibody for capturing the secretory substance contained in the first capturing part 101-1, shortening the distance between the image sensor 1001 and the first capturing part 101-1 (e.g., the antibody layer) as much as possible (disposing it directly above the image sensor 1001), increasing the exposure time, separating the background noise part from the signal part by image processing, etc. Furthermore, the acquisition of cell images and the detection of secretory substances can be performed in real time on the image sensor 1001.

[0041] Next, refer to Fig. 2. The cell evaluation device 2 includes an antibody layer (first capture unit) 201 including an antibody (first antibody) 200, a CMOS image sensor 2001 which is an image acquisition unit, a driver 2002, and a device 2003 including an analysis unit and a display unit. Note that in Fig. 2, the CMOS image sensor 2001 is illustrated with three pixels.

[0042] The CMOS image sensor 2001 is composed of a protective layer 202, a semiconductor substrate 203, and a wiring layer 204, in that order from the light incident side. A photodiode (PD) (three photodiodes (PD) in FIG. 2) is formed on the semiconductor substrate 203. An antibody layer 201 is disposed directly above the protective layer 202 (above the protective layer in FIG. 2). By disposing the antibody layer 201 directly above the protective layer 202, the CMOS image sensor 2001 can efficiently receive light emitted by a luminescent substance bound to a secretory substance captured by the antibody 200 contained in the antibody layer 201, and cell observation light. The cell observation light means light obtained by projecting natural light or indoor light onto the cells, for example.

[0043] 2 shows four wells (two in the left-right direction and two in the up-down direction in FIG. 2), but this means that in the cell evaluation device 2, four wells are arranged directly above one CMOS image sensor 2001 (protective layer 205 provided in the CMOS image sensor 2001). In the well, for example, an antibody 200 that captures a secretory substance is immobilized on a collagen coat (protective part), which traps pretreated cells, which are stimulated and cultured, and which releases a secretory substance from the cells, which is then captured by the antibody 200.

[0044] Further, refer to Fig. 3. The cell evaluation device 3 includes an antibody layer (first capture unit) 301 including an antibody 300, a CMOS image sensor 3001 which is an image acquisition unit, a driver 3002, a device 3003 including an analysis unit and a display unit, and an irradiation unit 3004. Note that in Fig. 3, three pixels of the CMOS image sensor 3001 are illustrated.

[0045] The CMOS image sensor 3001 is composed of, in order from the light incident side, a protective layer 302, a semiconductor substrate 303, and a wiring layer 304. A photodiode (PD) (three photodiodes (PD) in FIG. 3) is formed on the semiconductor substrate 303. An antibody layer 301 is disposed directly above the protective layer 302 (above the protective layer in FIG. 3). By disposing the antibody layer 301 directly above the protective layer 302, the CMOS image sensor 3001 can efficiently receive light emitted by a luminescent substance bound to a secretory substance captured by the antibody 300 contained in the antibody layer 301, and light obtained by projecting the irradiation unit 3004 onto a cell.

[0046] In FIG. 3, four wells (four in the vertical direction in FIG. 3) are shown, but in the cell evaluation device 3, this means that one well is placed directly above one CMOS image sensor 3001 (protective layer 305 provided in the CMOS image sensor 3001). In other words, at least four CMOS image sensors 3001 are required in the cell evaluation device 3. In the well, for example, an antibody 300 that captures a secretory substance is immobilized on a collagen coat (protective part), which traps pretreated cells, which are stimulated and cultured, and secretory substances are released from the cells, and the secretory substances are captured by the antibody 300.

[0047] [2-2. First capture unit] The cell evaluation device according to the first embodiment of the present technology includes a first capture unit. As described above, the first capture unit captures a secretory substance secreted by a cell. The first capture unit may include a molecule that binds non-specifically or specifically to the secretory substance. The molecule that binds non-specifically or specifically to the secretory substance may be at least one selected from the group consisting of a first antibody, a first aptamer, and an imprinted polymer, and the first antibody may be of multiple types. A protective unit may be disposed in an area of ​​the first capture unit that does not include a molecule that binds non-specifically or specifically to the secretory substance. As described above, the protective unit may be formed of, for example, a collagen coat. The protective unit may also be formed of at least one hydrophilic polymer that can suppress the adsorption of a biopolymer. Examples of hydrophilic polymers include polyethylene glycol, polymers having phosphorylcholine groups in their side chains, polysaccharides, and polypeptides.

[0048] [2-3. Image acquisition section] The cell evaluation device according to the first embodiment of the present technology includes an image acquisition unit. The image acquisition unit may be, for example, an image sensor, and the image sensor may be, for example, a solid-state imaging element such as a CMOS (Complementary Metal Oxide Semiconductor) image sensor or a CCD (Charge Coupled Device). Below, a back-illuminated solid-state imaging element and a front-illuminated solid-state imaging element, which are examples of a CMOS image sensor, will be described in detail.

[0049] (Back-illuminated solid-state imaging element) A back-illuminated solid-state imaging element, which is an example of a CMOS image sensor, will be described with reference to Fig. 4. Fig. 4 is a cross-sectional view showing an example of the configuration of one pixel 20 of a back-illuminated solid-state imaging element 10.

[0050] Each pixel 20 is configured to have one organic photoelectric conversion element 41, and photodiodes 36 and 37 having pn junctions, which are stacked in the depth direction within one pixel. A protective layer 44 is formed on the organic photoelectric conversion element 41 on the back side (upper side in FIG. 4) of the semiconductor substrate 35, and an antibody layer 50 containing an antibody 51 is formed directly on the protective layer 44. That is, the protective layer 44 and the antibody layer 50 are continuously arranged. In this case, the thickness of the protective layer 44 may be a thin layer of, for example, 100 nm. The protective layer 44, which is continuously arranged with the antibody layer 50, is preferably transparent in order to efficiently capture light into the organic photoelectric conversion element 41, the photodiode 36, and the photodiode 37. The protective layer 44 is preferably made of, for example, SiO2. The pixel 20 has a semiconductor substrate (silicon substrate) 35 on which a photodiode 36 and a photodiode 37 are formed, a light receiving surface where light is incident is formed on the back side of the semiconductor substrate 35 (the upper side of the semiconductor substrate 35 in FIG. 4), and a circuit including a readout circuit and the like is formed on the front side of the semiconductor substrate 35. That is, the pixel 20 has a light receiving surface on the back side of the substrate 35, and a circuit forming surface formed on the front side of the substrate opposite to the light receiving surface. The semiconductor substrate 35 may be composed of a semiconductor substrate of a first conductivity type, for example, an n-type.

[0051] In the semiconductor substrate 35, inorganic photoelectric conversion units having two pn junctions, i.e., a first photodiode 36 and a second photodiode 37, are formed so as to be stacked in the depth direction from the back surface side. In the semiconductor substrate 35, the first photodiode 36 is formed from the back surface toward the depth direction (downward in the figure), and the second photodiode 37 is formed. In Fig. 4, the first photodiode 36 is for blue (B), and the second photodiode 37 is for red (R).

[0052] An organic photoelectric conversion element 41 for a first color is disposed on the upper part of the rear surface of the semiconductor substrate 35 in the region where the first photodiode 36 and the second photodiode 37 are formed, the organic photoelectric conversion element 41 being formed by stacking a first electrode (lower electrode) 33, a first buffer layer 47, a photoelectric conversion layer 32, a second buffer layer 46, and a second electrode (upper electrode) 31 in this order. In the example of the back-illuminated solid-state imaging element 10 shown in Fig. 4, the organic photoelectric conversion element 41 is for green (G). The second electrode (upper electrode) 31 and the first electrode (lower electrode) 33 may be formed of a transparent conductive film such as an indium tin oxide film or an indium zinc oxide film.

[0053] 4, the organic photoelectric conversion element 41 is green, the first photodiode 36 is blue, and the second photodiode 37 is red, but other color combinations are possible. For example, the organic photoelectric conversion element 41 can be red or blue, and the first photodiode 36 and the second photodiode 37 can be set to other corresponding colors. In this case, the positions of the first photodiode 36 and the second photodiode 37 in the depth direction are set according to the color.

[0054] Also, without using the first photodiode 36 and the second photodiode 37, three photoelectric conversion elements, i.e., the blue organic photoelectric conversion element 41B, the green organic photoelectric conversion element 41G, and the red organic photoelectric conversion element 41R, may be applied to the solid-state imaging element (back-illuminated solid-state imaging element and front-illuminated solid-state imaging element) of the second embodiment according to the present technology. As the organic photoelectric conversion element 41B that performs photoelectric conversion with blue wavelength light, an organic photoelectric conversion material containing a coumarin-based dye, tris-8-hydroxyquinoline A1 (A1q3), a melacyanine-based dye, or the like may be used. As the photoelectric conversion element 41G that performs photoelectric conversion with green wavelength light, an organic photoelectric conversion material containing, for example, a rhodamine-based dye, a melacyanine-based dye, a quinacridone, or the like may be used. As the photoelectric conversion element 41R that performs photoelectric conversion with red wavelength light, an organic photoelectric conversion material containing a phthalocyanine-based dye may be used.

[0055] Furthermore, in addition to the blue organic photoelectric conversion element 41B, the green organic photoelectric conversion element 41G, and the red organic photoelectric conversion element 41R, an ultraviolet organic photoelectric conversion element 41UV and / or an infrared organic photoelectric conversion element 41IR may be applied to the solid-state imaging element (back-illuminated solid-state imaging element and front-illuminated solid-state imaging element) of the second embodiment according to the present technology. By providing the ultraviolet photoelectric conversion element 41UV and / or the infrared photoelectric conversion element 41IR, it becomes possible to detect light of a wavelength other than the visible light region.

[0056] The light emitted by the luminescent substance bound to the secretory substance captured by the antibody 51 (first antibody) contained in the antibody layer 50, the observation light of the cell, and the projection light of the cell projected by the light of the irradiation unit may be light having any wavelength band, and may be imaged using at least one of the organic photoelectric conversion element 41, the first photodiode 36, and the second photodiode 37 according to the color (wavelength band) of these lights. For example, in the solid-state imaging element 10, if the light emitted by the luminescent substance bound to the secretory substance is green light, it can be imaged using the organic photoelectric conversion element 41, if the light emitted by the luminescent substance bound to the secretory substance is blue light, it can be imaged using the first photodiode 36, and if the light emitted by the luminescent substance bound to the secretory substance is red light, it can be imaged using the first photodiode 37. Furthermore, for example, if the observation light for the cell or the light obtained by projecting the cell using the illumination unit is achromatic light (e.g., white light, gray light, etc.), imaging can be performed using all of the organic photoelectric conversion element 41, the first photodiode 36, and the second photodiode 37.

[0057] In the organic photoelectric conversion element 41, a first electrode (lower electrode) 33 is formed, and an insulating film 34 for insulating and isolating the first electrode (lower electrode) 33 is formed below the first electrode (lower electrode) 33 in the figure.

[0058] In one pixel 20, a wiring 39 connected to the first electrode (lower electrode) 33 and a wiring (not shown) connected to the second electrode (upper electrode) 31 are formed. The wiring 39 and the wiring connected to the second electrode (upper electrode) 31 can be formed, for example, from a tungsten (W) plug having a SiO2 or SiN insulating layer around it to suppress short circuiting with Si, or a semiconductor layer formed by ion implantation. In the example of the back-illuminated solid-state imaging element shown in FIG. 2, since the signal charge is a hole, the wiring 39 becomes a p-type semiconductor layer when formed from a semiconductor layer formed by ion implantation. The wiring connected to the second electrode (upper electrode) 31 can be an n-type semiconductor layer because the second electrode (upper electrode) 31 draws out electrons.

[0059] In this example, an n-type region 38 for storing electric charges is formed on the surface side of a semiconductor substrate 35. This n-type region 38 functions as a floating diffusion portion of a photoelectric conversion element 41.

[0060] A film having a negative fixed charge can be used as the insulating film 34 on the back surface of the semiconductor substrate 35. For example, a hafnium oxide film can be used as the film having a negative fixed charge. That is, the insulating film 34 may be formed to have a three-layer structure in which a silicon oxide film, a hafnium oxide film, and a silicon oxide film are formed in this order from the back surface.

[0061] A wiring layer 45 is formed on the front surface side (lower side in FIG. 4) of the semiconductor substrate 35. Although not shown, the back-illuminated solid-state imaging element 10 may be provided with a color filter, a transparent filter, an ND filter, a white filter, a gray filter, or the like, or may be provided with an on-chip lens.

[0062] (Front-illuminated solid-state imaging element) The CMOS image sensor may be not only the back-illuminated solid-state imaging element described above, but also a front-illuminated solid-state imaging element. A front-illuminated solid-state imaging element will now be described.

[0063] The example of the front-illuminated solid-state imaging element differs from the above-described back-illuminated solid-state imaging element 10 only in that the wiring layer 92 formed under the semiconductor substrate 35 is replaced by a wiring layer 92 formed between the organic photoelectric conversion element 41 and the semiconductor substrate 35. Other points may be the same as those of the above-described back-illuminated solid-state imaging element 10, and therefore a description thereof will be omitted here.

[0064] [2-4.Analysis Department] As described above, the cell evaluation device of the first embodiment according to the present technology can include an analysis unit. The analysis unit can quantify the amount of the secretory substance based on the intensity of light emitted from the luminescent substance bound to the secretory substance. The analysis unit can also analyze the cells based on the observation light for the cells or the light (brightness information) obtained by projecting the cells using the irradiation unit. The analysis unit may convert the projected image of the cells into a higher resolution image using hologram technology, super-resolution technology, or the like, and analyze the cells.

[0065] A specific configuration of the analysis unit will be described with reference to Fig. 5. Fig. 5 shows an example of the configuration of the analysis unit, and in detail, Fig. 5(a) shows antibody arrangement information 70a, Fig. 5(b) shows a chemiluminescence image, and Fig. 5(c) shows a cell image.

[0066] 5(a), antibody arrangement information 70a shows antibodies 71a to 76a. Each of antibodies 71a to 76a includes, for example, a first antibody that captures a secretory substance. In the case of a cell evaluation device according to a first embodiment of the present technology described below, each of antibodies 71a to 76a includes, for example, a first antibody that captures a secretory substance and a second antibody that captures a cell.

[0067] Referring to FIG. 5(b), individual chemiluminescent images 71b to 76b are obtained from the chemiluminescent image 70b. The chemiluminescent images 71b to 76b correspond to the antibodies 71a to 76a, respectively. The chemiluminescent intensity is calculated from the brightness information of the antibody arrangement areas of the chemiluminescent images 71b to 76b, and the amount of the secretion substance can be further calculated. Specifically, the quantification of the secretion substance is performed by obtaining the chemiluminescence intensity from the brightness information of each pixel obtained from the image sensor, and calculating the concentration of the secretion substance from the intensity and information of a calibration curve previously obtained as data.

[0068] Referring to FIG. 5(c), the cell image 70c shows individual cell images 71c-76c. In FIG. 5(c), the cell images 71c-76c are cell images at 1x (actual size) of the cells, but may be enlarged or reduced cell images as necessary. Since the cell images 71c-76c correspond to the antibodies 71a-76a and the chemiluminescent images 71b-76b, respectively, the concentration of the secretion substance for each of the cell images 71c-76c can be uniquely determined. Then, object segmentation is performed from the luminance information of the cell images 71c-76c imaged on the image sensor by the cell observation light or irradiation unit, and the size, position and shape of each of the cell images 71c-76c can be analyzed, and the internal state of the cell can be analyzed from the dynamic range within the cell, and further the total number of cells contained in one well can be calculated. For example, the size of the cell shown as the cell image 71c corresponds to the arrow P7. In the case of a cell evaluation device according to a second embodiment of the present technology described later, each of the cells in the cell images 71c to 76c is a cell captured by a second antibody, and therefore the positional information of the cell becomes clearer.

[0069] [2-5. Irradiation unit] As described above, the cell evaluation device of the first embodiment according to the present technology can include an irradiation unit. The irradiation unit is one means for confirming a cell image, and as described above, the wavelength selectivity (color selectivity of light) of the light emitted by the irradiation unit and the exposure time of the light can be controlled using the irradiation control unit, so that a clearer cell image can be obtained by using the irradiation unit. In addition, the photodegradable linker can be cleaved by utilizing the specific wavelength of the light irradiated, and only specific cells can be released.

[0070] [2-6.Display section] The cell evaluation device according to the first embodiment of the present technology may include a display unit. The display unit may include a well image display unit based on a cell image obtained by capturing light (brightness information) obtained by projecting a cell using observation light for the cell or an irradiation unit and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to a secretory substance, and an analysis data display unit and an analysis plot display unit based on the analysis by the analysis unit.

[0071] The specific configuration of the display unit will be described with reference to Fig. 6. Fig. 6 shows an example of the configuration of the display unit, and in detail, Fig. 6(b) shows a well image display unit based on the chemiluminescence image 80a-1 and cell image image 80a-2 shown in Fig. 6(a), Fig. 6(c) shows an analysis data display unit, and Fig. 6(c) shows an analysis plot display unit.

[0072] 6(a) and (b), the well display section 80b displays an image obtained by synthesizing each of the chemiluminescent images 81a-1 to 86a-1 of the chemiluminescent image 80a-1 with each of the cell images 81a-2 to 86a-2 in correspondence with each other. In FIG. 6(b), information on the cells contained in the well 1A is displayed among the total of six wells, 1A to 3A and 2A to 3B. In FIG. 6(b), the chemiluminescent image 81b-1 corresponds to the cell image 81b-2, the chemiluminescent image 82b-1 corresponds to the cell image 82b-2, the chemiluminescent image 83b-1 corresponds to the cell image 83b-2, the chemiluminescent image 84b-1 corresponds to the cell image 84b-2, the chemiluminescent image 85b-1 corresponds to the cell image 85b-2, and the chemiluminescent image 86b-1 corresponds to the cell image 86b-2. Among these, the cell in cell image 81a-2 (cell image 81b-2), whose cell size is indicated by arrow P8, is the largest of the six cells shown in FIG. 6(b), and has the highest amount of chemiluminescence. In this case, the cell is shown in white as a chemiluminescence image in FIGS. 6(a) and (b). In contrast, the cell in cell image 83a-2 (cell image 83b-2) is the smallest, so small that it is not shown, and has an amount of chemiluminescence that is almost zero. In this case, the cell is shown in black as a chemiluminescence image in FIGS. 6(a) and (b).

[0073] Referring to FIG. 6(c), the analysis data display section 80c shows that the total number of cells in well 1A is 10 4 The number of cells is indicated, and the cell diameter (μm), secretion amount (pg), etc. of each cell (cell number) are shown.

[0074] Referring to Fig. 6(d), the analysis plot display section 80d shows a graph in which the vertical axis is the secretion amount (pg) and the horizontal axis is the cell size (cell diameter) (µm). Referring to the graph shown in Fig. 6(d), judging from the correlation between the secretion amount (pg) and the cell size (cell diameter) (µm), the number of cells to be evaluated is 10 4It can be determined that there are 100 out of 100 cells. In Fig. 6(d), the cells indicated by black circles out of the 100 cells correspond to, for example, the cells in the cell image 81a-2 (cell image 81b-2) described above.

[0075] 3. Second embodiment (Example 2 of cell evaluation device) [3-1. Cell evaluation device] The cell evaluation device according to the second embodiment (example 2 of the cell evaluation device) of the present technology includes a first capture unit that captures a secretory substance secreted by a cell, a second capture unit that captures a cell, and an image acquisition unit that captures an observation light of the cell and light emitted from a luminescent substance bound to the secretory substance to acquire an image, in which the image acquisition unit and the first capture unit are arranged in this order, and further, the image acquisition unit and the second capture unit are arranged in this order, and the image acquisition unit acquires position information of the cell and the secretory substance. That is, the cell evaluation device according to the second embodiment (example 2 of the cell evaluation device) of the present technology has a configuration in which the second capture unit is added to the cell evaluation device according to the first embodiment (example 1 of the cell evaluation device) of the present technology, and the configuration of the cell evaluation device according to the second embodiment (example 2 of the cell evaluation device) of the present technology may be the same as the configuration of the cell evaluation device according to the first embodiment (example 1 of the cell evaluation device) of the present technology, except for the configuration of the second capture unit. Here, the image acquisition section and the first capture section being arranged in this order means that the image acquisition section and the first capture section may be arranged continuously, or the image acquisition section and the first capture section may be arranged via an optional material layer (material film) or material section. The image acquisition section and the first capture section being arranged continuously means that the first capture section is arranged directly above the image acquisition section. The first capture section being arranged directly above the image acquisition section means, for example, that the first capture section is arranged at a distance of 150 μm or less, preferably 10 μm or less, from the image acquisition section.

[0076] Moreover, the image acquisition section and the second capture section being arranged in this order means that the image acquisition section and the second capture section may be arranged continuously, or the image acquisition section and the second capture section may be arranged via an optional material layer (material film) or material section. The image acquisition section and the second capture section being arranged continuously means that the second capture section is arranged directly above the image acquisition section. The second capture section being arranged directly above the image acquisition section means, for example, that the second capture section is arranged at a distance of 150 μm or less, preferably 10 μm or less, from the image acquisition section.

[0077] The cell evaluation device of the second embodiment according to the present technology may further include at least one of an analysis unit, an irradiation unit, and a display unit. That is, the cell evaluation device of the second embodiment according to the present technology may further include an analysis unit, an irradiation unit, a display unit, an analysis unit and an irradiation unit, an analysis unit and a display unit, an irradiation unit and a display unit, or an analysis unit, an irradiation unit, and a display unit.

[0078] Please refer to FIG. 7. The cell evaluation device 7 includes at least an upper part 101 of the image sensor including a first capture unit 101-1 and a second capture unit 101-2, an image sensor 1001 (e.g., a CMOS image sensor) which is an image acquisition unit, and an analysis unit 1005. The cell evaluation device 1 further includes a driver 1002, an irradiation control unit 1004, a display unit 1006, and an irradiation unit 1007. As shown in FIG. 1, the irradiation control unit 1004, the analysis unit 1005, and the display unit 1006 may be formed as one device 1003. Although not shown in FIG. 7, the upper part 101 of the image sensor may include a protection unit in addition to the first capture unit 101-1 and the second capture unit 101-2.

[0079] In the cell evaluation device 7, a first capture unit 101-1 (e.g., a capture unit including a first antibody for capturing a secretory substance) and a second capture unit 101-2 (e.g., a capture unit including a second antibody for capturing a cell) are arranged in a desired positional relationship on an image sensor 1001. The cells captured by the second capture unit 101-2 can be identified by taking a cell image using a cell observation light or irradiation unit 1007, and analyzing the cell size and cell shape by image analysis. The cell type and cell state (such as cell life and death) can be accurately grasped. Since the cells are captured by the second capture unit 101-2, it is also possible to recover the cells by cutting only a specific capture antibody (second antibody) that reacts to a secretory substance, for example, by a photolinker technique, after the cells are cultured. In this case, it is preferable that a photodegradable linker is included between the second capture unit 101-2 and the image sensor 1001. By including a photodegradable linker, only specific cells can be released. That is, in the cell evaluation device 7, after analyzing the cells, the cell capture is terminated by irradiating the cells with light, and only the desired cells are taken out, making it possible to perform post-analysis (genetic analysis, etc.).

[0080] Then, cells are cultured on the image sensor 1001, and a secretory substance released from the cells is captured by the first capture unit 101-1 (for example, a capture unit including a first antibody for capturing the secretory substance), and then further reacted with a chemiluminescent substance or the like to generate weak light, which can be acquired by the image sensor 1001 directly below. The chemiluminescent substance is, for example, an enzyme, and chemiluminescence is emitted by the reaction of the enzyme with a substrate.

[0081] The weak light is photoelectrically converted into an electric signal by the driver 1002, and the brightness of only the sensor part where the secretory substance is captured increases, so that it is possible to map the position where the secretory substance is obtained on the image sensor 1001. Since the positional relationship between the first capture part 101-1 (e.g., a capture part including a first antibody for capturing the secretory substance) and the second capture part 101-2 (e.g., a capture part including a second antibody for capturing the cell) can be specified in advance, it is also possible to identify the cell that released the secretory substance. In addition, the analysis part 1005 can also quantify the secretory substance by utilizing the fact that the brightness changes depending on the concentration of the secretory substance. The image obtained from the image sensor 1001 is displayed on the display part 1006 (e.g., a monitor) through the analysis part 1005.

[0082] In order to increase the sensitivity, there are measures such as increasing the density of the first antibody for capturing the secretory substance contained in the first capturing part 101-1, shortening the distance between the image sensor 1001 and the first capturing part 101-1 (e.g., the antibody layer) as much as possible (disposing it directly above the image sensor 1001), increasing the exposure time, separating the background noise part from the signal part by image processing, etc. Furthermore, the acquisition of cell images and the detection of the secretory substance can be performed in real time on the image sensor 1001.

[0083] [3-2. First capture portion and second capture portion, and patterning of the first capture portion and the second capture portion] The cell evaluation device of the second embodiment of the present technology includes a first capture unit and a second capture unit. As described above, the first capture unit captures a secretory substance secreted by a cell. The first capture unit is as described in the section on the cell evaluation device of the first embodiment of the present technology (example 1 of the cell evaluation device), so a detailed description will be omitted here.

[0084] The second capture part captures cells. The second capture part may contain a molecule that binds non-specifically or specifically to cells. The molecule that binds non-specifically or specifically to cells may be at least one selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine, and the second antibody may be of multiple types. A protective part may be disposed in an area of ​​the second capture part that does not contain a molecule that binds non-specifically or specifically to cells. The protective part may be formed, for example, from a collagen coat. The protective part may also be formed from at least one hydrophilic polymer that can suppress the adsorption of biopolymers. Examples of hydrophilic polymers include polyethylene glycol, polymers having phosphorylcholine groups in their side chains, polysaccharides, and polypeptides.

[0085] The patterning of the first capture portion and the second capture portion will be described with reference to Figures 8 to 10. Referring to Figure 8(a), in a size of 50 μm, the first capture portion 402a is disposed around the second capture portion 401a, and a protective portion 403a is disposed between the first capture portion 402a and the second capture portion 401a. Figure 8(b) shows 16 patterns of the first capture portion and the second capture portion shown in Figure 8(a). The second capture portion 401a shown in Figure 8(a) corresponds to the second capture portion 401b shown in Figure 8(b), the first capture portion 402a shown in Figure 8(a) corresponds to the second capture portion 402b shown in Figure 8(b), and the protection portion 403a shown in Figure 8(a) corresponds to the second protection portion 403b shown in Figure 8(b).

[0086] Since the cell evaluation device of the first embodiment (example 1 of the cell evaluation device) of the present technology does not include the second capturing unit 401a, the first capturing unit 402a is disposed in place of the second capturing unit 401a.

[0087] 9(a), in a size of 50 μm, first capturing parts 502a-505a are arranged around second capturing part 501a, and protective parts 506a are arranged between first capturing parts 502a-505a and second capturing part 501a and between first capturing parts 502a-505a. The first capturing parts 502a and 503a contain the same material (e.g., a first antibody), and the first capturing parts 503a and 504a contain the same material (e.g., a first aptamer), and thus two kinds of materials are used. In Fig. 9(a), the first capture parts 502a and 503a are vertically rectangular (the longitudinal direction is the up-down direction in Fig. 9(a)), and the first capture parts 504a and 505a are horizontally rectangular (the longitudinal direction is the left-right direction in Fig. 9(a)), so that the first capture parts containing different materials are adjacent to each other. Fig. 9(b) shows 16 patterns of one first capture part and one second capture part shown in Fig. 9(a).

[0088] The second capture portion 501a shown in Figure 9(a) corresponds to the second capture portion 501b shown in Figure 9(b), the first capture portion 502a shown in Figure 9(a) corresponds to the second capture portion 502b shown in Figure 9(b), the first capture portion 503a shown in Figure 9(a) corresponds to the second capture portion 503b shown in Figure 9(b), the first capture portion 504a shown in Figure 9(a) corresponds to the second capture portion 504b shown in Figure 9(b), the first capture portion 505a shown in Figure 9(a) corresponds to the second capture portion 505b shown in Figure 9(b), and the protective portion 506a shown in Figure 9(a) corresponds to the protective portion 506b shown in Figure 9(b).

[0089] Since the cell evaluation device of the first embodiment (example 1 of the cell evaluation device) of the present technology does not include a second capture unit 501a, at least one of the first capture units 502a to 505a is arranged in place of the second capture unit 501a.

[0090] Referring to FIG. 10(a), in a size of 50 μm, the first capture parts 602a-609a are arranged around the second capture part 601a, and protective parts 610a are arranged between the first capture parts 602a-609a and the second capture part 601a, and between each of the first capture parts 602a-609a. Each of the first capture parts 602a-609a includes different materials (for example, eight types of first antibodies or eight types of first aptamers), and eight types of materials are used. The first capture parts 602a-609a are arranged in eight square shapes in FIG. 10(a). FIG. 10(b) shows 16 patterns of one first capture part and one second capture part shown in FIG. 10(a).

[0091] 10(a) corresponds to the second trapping portion 601(b) shown in FIG. 10(b), the first trapping portion 602a shown in FIG. 10(a) corresponds to the second trapping portion 602b shown in FIG. 10(b), the first trapping portion 603a shown in FIG. 10(a) corresponds to the second trapping portion 603b shown in FIG. 10(b), the first trapping portion 604a shown in FIG. 10(a) corresponds to the second trapping portion 604b shown in FIG. 10(b), and the first trapping portion 605a shown in FIG. 10(a) corresponds to the second trapping portion 605b shown in FIG. 10(a) corresponds to the second capture portion 606b shown in FIG. 10(b), the first capture portion 607a shown in FIG. 10(a) corresponds to the second capture portion 607b shown in FIG. 10(b), the first capture portion 608a shown in FIG. 10(a) corresponds to the second capture portion 608b shown in FIG. 10(b), the first capture portion 609a shown in FIG. 10(a) corresponds to the second capture portion 609b shown in FIG. 9(b), and the protective portion 610a shown in FIG. 10(a) corresponds to the protective portion 610b shown in FIG. 9(b). Since the cell evaluation device of the first embodiment (example 1 of the cell evaluation device) of the present technology does not include a second capture unit 601a, at least one of the first capture units 602a to 609a is arranged in place of the second capture unit 601a.

[0092] [3-3. Image acquisition unit, analysis unit, irradiation unit and display unit] The image acquisition unit included in the cell evaluation device of the second embodiment (example 2 of the cell evaluation device) according to the present technology is as described in the section on the cell evaluation device of the first embodiment (example 1 of the cell evaluation device) according to the present technology, and therefore a detailed description thereof will be omitted here. In addition, the analysis unit, irradiation unit, and display unit that can be included in the cell evaluation device of the second embodiment (example 2 of the cell evaluation device) according to the present technology are as described in the section on the cell evaluation device of the first embodiment (example 1 of the cell evaluation device) according to the present technology, and therefore a detailed description thereof will be omitted here.

[0093] <4. Third embodiment (example of cell evaluation system)> [4-1. Cell evaluation system] The cell evaluation system of a third embodiment (an example of a cell evaluation system) of the present technology includes a first capture unit that captures a secretory substance secreted by a cell, an image acquisition unit that acquires an image by capturing the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance, and an analysis unit that analyzes the acquired image, in which the image acquisition unit and the first capture unit are arranged in this order, and the image acquisition unit acquires positional information of the cell and the secretory substance.

[0094] Furthermore, a cell evaluation system according to a third embodiment (an example of a cell evaluation system) of the present technology may be a cell evaluation system including a first capture unit that captures a secretory substance secreted by a cell, an image acquisition unit that acquires an image obtained by capturing observation light for the cell and light emitted from a luminescent substance bound to the secretory substance, and an analysis unit that analyzes the acquired image, in which the image acquisition unit and the first capture unit are arranged in this order, and the image acquisition unit acquires positional information of the cell and the secretory substance.

[0095] The cell evaluation system of the third embodiment according to the present technology may further include at least one of an irradiation unit and a display unit. That is, the cell evaluation system of the third embodiment according to the present technology may further include an irradiation unit, may include a display unit, or may include an irradiation unit and a display unit.

[0096] In addition, with respect to the cell evaluation system of the third embodiment of the present technology, other than the contents described above, the contents described in the sections on the cell evaluation devices of the first and second embodiments of the present technology (Examples 1 and 2 of the cell evaluation device) can be directly applied to the cell evaluation system of the third embodiment of the present technology.

[0097] [4-2. First capture unit, second capture unit, image acquisition unit, analysis unit, irradiation unit, and display unit] The image acquisition unit and analysis unit provided in the cell evaluation system of the third embodiment (cell evaluation system) of the present technology are as described in the section on the cell evaluation device of the first embodiment (example 1 of the cell evaluation device) of the present technology, so detailed description will be omitted here. In addition, the irradiation unit and display unit that can be provided in the cell evaluation system of the third embodiment (cell evaluation system) of the present technology are as described in the section on the cell evaluation device of the first embodiment (example 1 of the cell evaluation device) of the present technology, so detailed description will be omitted here. Furthermore, the second capture unit that can be provided in the cell evaluation system of the third embodiment (cell evaluation system) of the present technology is as described in the section on the cell evaluation device of the second embodiment (example 2 of the cell evaluation device) of the present technology, so detailed description will be omitted here.

[0098] Note that the embodiments according to the present technology are not limited to the above-described embodiments, and various modifications are possible without departing from the gist of the present technology.

[0099] Furthermore, the effects described in this disclosure are merely examples and are not limiting, and other effects may also be present.

[0100] This technology can also take the following configurations [1] to

[68] . [1] a first capture unit that captures a secretory substance secreted by a cell; and an image acquisition unit that acquires an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance, the image acquisition unit and the first capture unit are arranged in this order, The image acquisition unit acquires position information of the cell and the secretory substance. [2] The cell evaluation device according to [1], wherein the image acquisition unit is a CMOS image sensor. [3] The cell evaluation device according to [1] or [2], wherein the image acquisition unit acquires an image by capturing chemiluminescence emitted by the luminescent substance, which is an enzyme, upon reaction with a substrate. [4] The cell evaluation device according to any one of [1] to [3], wherein the first capture portion includes a molecule that binds non-specifically or specifically to the secretory substance. [5] The cell evaluation device according to [4], wherein a protective portion is disposed in an area of ​​the first capture portion that does not contain the molecule that binds non-specifically or specifically to the secreted substance. [6] The cell evaluation device described in [4] or [5], wherein the molecule that non-specifically or specifically binds to the secreted substance is at least one selected from the group consisting of a first antibody, a first aptamer, and a molecularly imprinted polymer. [7] The cell evaluation device according to [6], wherein the first antibody is of multiple types.

[0101] [8] Further comprising a second capture unit that captures the cells; The cell evaluation device described in any one of [1] to [7], wherein the image acquisition unit and the second capture unit are arranged in this order. [9] The cell evaluation device described in [8], wherein the first capture portion is arranged around the second capture portion.

[10] The cell evaluation device described in [8] or [9], wherein a photodegradable linker is included between the second capture unit and the image acquisition unit.

[11] The cell evaluation device described in any one of [8] to

[10] , wherein the second capture portion includes a molecule that binds non-specifically or specifically to the cell.

[12] The cell evaluation device described in

[11] , wherein a protective portion is arranged in an area of ​​the second capture portion that does not contain the molecule that binds non-specifically or specifically to the cell.

[13] The cell evaluation device according to

[11] or

[12] , wherein the molecule that non-specifically or specifically binds to the cell is at least one selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine.

[14] The cell evaluation device according to

[13] , wherein the second antibody is of multiple types.

[0102]

[15] a first capture unit that captures a secretory substance secreted by a cell; an image acquisition unit that acquires an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance; An analysis unit that analyzes the obtained image, the image acquisition unit and the first capture unit are arranged in this order, The image acquisition unit acquires position information of the cell and the secretory substance.

[16] The cell evaluation device according to

[15] , wherein the analysis unit quantifies the secretory substance based on the intensity of the light emitted from the luminescent substance bound to the secretory substance.

[17] The cell evaluation device described in

[15] or

[16] , wherein the analysis unit analyzes the cells based on brightness information of the observation light of the cells and / or light emitted from a luminescent substance bound to a secreted substance.

[18] The cell evaluation device according to any one of

[15] to

[17] , wherein the image acquisition unit is a CMOS image sensor.

[19] The cell evaluation device described in any one of

[15] to

[18] , wherein the image acquisition unit acquires an image obtained by capturing chemiluminescence emitted by the luminescent substance, which is an enzyme, upon reaction with a substrate.

[20] The cell evaluation device according to any one of

[15] to

[19] , wherein the first capture portion includes a molecule that binds non-specifically or specifically to the secretory substance. [twenty one] The cell evaluation device described in

[20] , wherein a protective portion is arranged in an area of ​​the first capture portion that does not contain the molecule that non-specifically or specifically binds to the secreted substance. [twenty two] The cell evaluation device according to

[20] or

[21] , wherein the molecule that non-specifically or specifically binds to the secreted substance is at least one selected from the group consisting of a first antibody, a first aptamer, and a molecularly imprinted polymer. [twenty three] The cell evaluation device according to

[22] , wherein the first antibody is of multiple types.

[0103] [twenty four] Further comprising a second capture unit that captures the cells; The cell evaluation device described in any one of

[15] to

[23] , wherein the image acquisition unit and the second capture unit are arranged in this order. [twenty five] The cell evaluation device described in

[24] , wherein the first capture portion is arranged around the second capture portion.

[26] The cell evaluation device described in

[24] or

[25] , wherein a photodegradable linker is included between the second capture unit and the image acquisition unit.

[27] The cell evaluation device described in any one of

[24] to

[26] , wherein the second capture portion includes a molecule that binds non-specifically or specifically to the cell.

[28] The cell evaluation device described in

[27] , wherein a protective portion is arranged in an area of ​​the second capture portion that does not contain the molecule that binds non-specifically or specifically to the cell.

[29] The cell evaluation device described in

[27] or

[28] , wherein the molecule that non-specifically or specifically binds to the cell is at least one selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine.

[30] The cell evaluation device according to

[29] , wherein the second antibody is of multiple types.

[0104]

[31] The cell evaluation device described in any one of [1] to

[30] , further comprising an irradiation unit that projects the cells.

[32] The cell evaluation device described in any one of [1] to

[31] , further comprising a display unit for displaying the image.

[33] The cell evaluation device described in

[32] , wherein the display unit is provided with a well image display unit based on a cell image obtained by capturing an observation light of the cell and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to the secretory substance, as well as an analysis data display unit and an analysis plot display unit based on the analysis by the analysis unit.

[0105]

[34] a first capture unit that captures a secretory substance secreted by a cell; and an image acquisition unit that acquires an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance, the image acquisition unit and the first capture unit are arranged in this order, The image acquisition unit acquires position information of the cell and the secretory substance.

[35] The cell evaluation system described in

[34] , wherein the image acquisition unit is a CMOS image sensor.

[36] The cell evaluation system according to

[34] or

[35] , wherein the image acquisition unit acquires an image by capturing chemiluminescence emitted by the luminescent substance, which is an enzyme, upon reaction with a substrate.

[37] The cell evaluation system according to any one of

[34] to

[36] , wherein the first capture portion includes a molecule that binds non-specifically or specifically to the secreted substance.

[38] The cell evaluation system described in

[37] , wherein a protective portion is arranged in an area of ​​the first capture portion that does not contain the molecule that non-specifically or specifically binds to the secreted substance.

[39] The cell evaluation system described in

[37] or

[38] , wherein the molecule that non-specifically or specifically binds to the secreted substance is at least one selected from the group consisting of a first antibody, a first aptamer, and a molecularly imprinted polymer.

[40] The cell evaluation system described in

[39] , wherein the first antibody is of multiple types.

[0106]

[41] Further comprising a second capture unit that captures the cells; The cell evaluation system described in any one of

[34] to

[40] , wherein the image acquisition unit and the second capture unit are arranged in this order.

[42] The cell evaluation system described in

[41] , wherein the first capture portion is arranged around the second capture portion.

[43] The cell evaluation system described in

[41] or

[42] , wherein a photodegradable linker is included between the second capture unit and the image acquisition unit.

[44] The cell evaluation system according to any one of

[41] to

[43] , wherein the second capture portion includes a molecule that binds non-specifically or specifically to the cell.

[45] The cell evaluation system described in

[44] , wherein a protective portion is arranged in an area of ​​the second capture portion that does not contain the molecule that binds non-specifically or specifically to the cell.

[46] The cell evaluation system described in

[44] or

[45] , wherein the molecule that non-specifically or specifically binds to the cell is at least one selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine.

[47] The cell evaluation system described in

[46] , wherein the second antibody is of multiple types.

[0107]

[48] a first capture unit that captures a secretory substance secreted by a cell; an image acquisition unit that acquires an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance; An analysis unit that analyzes the obtained image, the image acquisition unit and the first capture unit are arranged in this order, The image acquisition unit acquires position information of the cell and the secretory substance.

[49] The cell evaluation system described in

[48] , wherein the analysis unit quantifies the secretory substance based on the intensity of light emitted from the luminescent substance bound to the secretory substance.

[50] The cell evaluation system described in

[48] or

[49] , wherein the analysis unit analyzes the cells based on brightness information of the observation light of the cells and / or light emitted from a luminescent substance bound to a secreted substance.

[51] The cell evaluation system according to any one of

[48] to

[50] , wherein the image acquisition unit is a CMOS image sensor.

[52] The cell evaluation system described in any one of

[48] to

[51] , wherein the image acquisition unit acquires an image by capturing chemiluminescence emitted by the luminescent substance, which is an enzyme, upon reaction with a substrate.

[53] The cell evaluation system according to any one of

[48] to

[52] , wherein the first capture portion includes a molecule that binds non-specifically or specifically to the secreted substance.

[54] The cell evaluation system described in

[53] , wherein a protective portion is arranged in an area of ​​the first capture portion that does not contain the molecule that non-specifically or specifically binds to the secreted substance.

[55] The cell evaluation system according to

[53] or

[54] , wherein the molecule that non-specifically or specifically binds to the secreted substance is at least one selected from the group consisting of a first antibody, a first aptamer, and a molecularly imprinted polymer.

[56] The cell evaluation system described in

[55] , wherein the first antibody is of multiple types.

[0108]

[57] Further comprising a second capture unit that captures the cells; The cell evaluation system described in any one of

[48] to

[56] , wherein the image acquisition unit and the second capture unit are arranged in this order.

[58] The cell evaluation system described in

[57] , wherein the first capture portion is arranged around the second capture portion.

[59] The cell evaluation system described in

[57] or

[58] , wherein a photodegradable linker is included between the second capture unit and the image acquisition unit.

[60] The cell evaluation system described in any one of

[57] to

[59] , wherein the second capture portion includes a molecule that binds non-specifically or specifically to the cell.

[61] The cell evaluation system described in

[60] , wherein a protective portion is arranged in an area of ​​the second capture portion that does not contain the molecule that binds non-specifically or specifically to the cell.

[62] The cell evaluation system described in

[60] or

[61] , wherein the molecule that non-specifically or specifically binds to the cell is at least one selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine.

[63] The cell evaluation system described in

[62] , wherein the second antibody is of multiple types.

[0109]

[64] The cell evaluation system described in any one of

[34] to

[63] , further comprising an irradiation unit that projects the cells.

[65] The cell evaluation system described in any one of

[34] to

[64] , further comprising a display unit for displaying the image.

[66] The cell evaluation system described in

[65] , wherein the display unit is provided with a well image display unit based on a cell image obtained by capturing an observation light of the cell and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to the secreted substance, as well as an analysis data display unit and an analysis plot display unit based on the analysis by the analysis unit.

[0110]

[67] a first antibody that captures a secretory substance secreted by a cell; a second antibody that captures the cells; and a CMOS image sensor for capturing an image of the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance, the CMOS image sensor, the first antibody, and the second antibody are arranged in this order; The first antibody is disposed around the second antibody, The CMOS image sensor acquires position information of the cells and the secretory substance.

[0111]

[68] a first antibody that captures a secretory substance secreted by a cell; a second antibody that captures the cells; a CMOS image sensor for capturing an image obtained by capturing the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance; An analysis unit that analyzes the acquired image, the CMOS image sensor, the first antibody, and the second antibody are arranged in this order; The first antibody is disposed around the second antibody, The CMOS image sensor acquires position information of the cells and the secretory substance.

[0112]

[69] a first antibody that captures a secretory substance secreted by a cell; a second antibody that captures the cells; and a CMOS image sensor for capturing an image of the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance, the CMOS image sensor, the first antibody, and the second antibody are arranged in this order; The first antibody is disposed around the second antibody, The CMOS image sensor acquires positional information of the cells and the secreted substances.

[0113]

[70] a first antibody that captures a secretory substance secreted by a cell; a second antibody that captures the cells; a CMOS image sensor for capturing an image obtained by capturing an observation light of the cell and a light emitted from a luminescent substance bound to the secretory substance; An analysis unit that analyzes the acquired image, the CMOS image sensor, the first antibody, and the second antibody are arranged in this order; The first antibody is disposed around the second antibody, The CMOS image sensor acquires positional information of the cells and the secreted substances. [Explanation of symbols]

[0114] 1...cell evaluation device, 101-1...first capture unit, 101-2...second capture unit, 1001...image acquisition unit (image sensor), 1005...analysis unit, 1006...display unit.

Claims

1. a cell capturing step of introducing a plurality of cells into a container having a first capturing part for capturing a secretory substance secreted by a cell and a second capturing part for capturing the cell disposed on a semiconductor substrate, and causing the second capturing part to capture at least one of the plurality of cells; a cell stimulating step of providing a stimulus to the plurality of cells; a secretory substance capturing step of causing the first capturing unit to capture a secretory substance secreted from at least one cell among the plurality of cells; an image acquiring step of acquiring an image obtained by capturing an observation light of at least one cell among the plurality of cells and a light emitted from a luminescent substance bound to the secretory substance; an analysis step of performing an analysis using the image obtained in the image acquisition step; Including, The image includes a cell image obtained by capturing the observation light, and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to the secretory substance, Each of the cell images in the cell image image corresponds to each of the chemiluminescence images in the chemiluminescence image; The analyzing step quantifies the secretory substance for each of the cells based on the correspondence. Cellular evaluation methods.

2. The image acquisition step acquires position information of the cells and the secretory substances. The cell evaluation method according to claim 1 .

3. The analyzing step analyzes at least one of the type, size, position, shape, and state of the cells. The cell evaluation method according to claim 1 .

4. The method further comprises a recovery step of recovering a desired cell from among the plurality of cells based on the information obtained in the analysis step. The cell evaluation method according to claim 1 .

5. The cell evaluation method according to claim 1 , wherein the semiconductor substrate is a semiconductor substrate on which a photodiode is formed.

6. The semiconductor substrate is a component of a CMOS sensor. The cell evaluation method according to claim 1 .

7. The image acquisition step is performed by the CMOS sensor. The cell evaluation method according to claim 6.

8. In the image acquisition step, a chemiluminescence image is obtained by capturing an image of chemiluminescence emitted by the reaction of the luminescent substance, which is an enzyme, with a substrate. The cell evaluation method according to claim 1 .

9. The first capture portion includes a molecule that non-specifically or specifically binds to the secreted substance. The cell evaluation method according to claim 1 .

10. The second capture portion includes a molecule that non-specifically or specifically binds to the cell. The cell evaluation method according to any one of claims 1 to 9.

11. The second capture portion comprises a photocleavable linker. The cell evaluation method according to any one of claims 1 to 10.

12. a container including a semiconductor substrate on which a first capture unit for capturing a secretory substance secreted by a cell and a second capture unit for capturing the cell are disposed; an image acquisition unit that acquires an image obtained by capturing light emitted from a light-emitting substance bound to the secretory substance captured in the first capture unit in the container and an observation light of the cell captured in the second capture unit; An analysis unit that analyzes the obtained image, The image includes a cell image obtained by capturing the observation light, and a chemiluminescence image obtained by capturing light emitted from a luminescent substance bound to the secretory substance, Each of the cell images in the cell image image corresponds to each of the chemiluminescence images in the chemiluminescence image; The analysis unit quantifies the secretory substance for each of the cells based on the correspondence. Cell evaluation system.

Images