Cell evaluation equipment and cell evaluation system

Abstract

Provided is a cell evaluation device capable of evaluating cells and substances secreted by cells with high precision and accuracy. The cell evaluation device includes a first capturing unit for capturing secretory substances secreted by cells and an image acquisition unit for acquiring an image obtained by imaging cell observation light and light emitted by a luminescent substance combined with the secretory substances. The image acquisition unit and the first capturing unit are arranged in the order of the image acquisition unit followed by the first capturing unit. The image acquisition unit acquires position information of cells and secretory substances.

Description

Technical Field

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

[0002] In recent years, systems for evaluating cells and secretory substances produced from cells have continued to develop, and various techniques have been proposed.

[0003] For example, a technology related to a complementary metal oxide semiconductor (CMOS) biosensor system has been proposed (see Patent Document 1).

[0004] Citation List

[0005] Patent Literature

[0006] Patent Document 1: U.S. Patent Application Publication No. 2012 / 0032235 Summary of the invention

[0007] Problems to be solved by the present invention

[0008] However, with the technique proposed in Patent Document 1, it may not be possible to accurately and precisely evaluate cells and secretory substances produced from the cells.

[0009] Therefore, the present technology has been achieved in view of such circumstances, and a main object of the present technology is to provide a cell evaluation device capable of accurately and precisely evaluating cells and secretory substances produced from cells.

[0010] Problem Solution

[0011] The present inventors have conducted intensive studies to solve the above-mentioned object. As a result, surprisingly, the present inventors have succeeded in being able to accurately and precisely evaluate cells and secretory substances produced from cells, and have completed the present technology.

[0012] That is, the present technology first provides a cell evaluation device, comprising:

[0013] a first capturing unit that captures secreted substances secreted by cells; and

[0014] an image acquisition unit that acquires an image obtained by imaging the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance, wherein

[0015] The image acquisition unit and the first capture unit are arranged in sequence, and

[0016] The image acquisition unit acquires position information of cells and secreted substances.

[0017] In the cell evaluation apparatus according to the present technology, the image acquisition unit may be a CMOS image sensor.

[0018] In the cell evaluation apparatus according to the present technology, the image acquisition unit acquires an image obtained by imaging chemiluminescence emitted by causing the luminescent substance as an enzyme to react with a substrate.

[0019] In the cell evaluation device according to the present technology, the first capture unit includes a molecule that non-specifically or specifically binds to the secretory substance.

[0020] In the cell evaluation apparatus according to the present technology, a protection unit is provided in a region where the first capture unit does not include the molecule that non-specifically or specifically binds to the secretory substance.

[0021] The molecule that non-specifically or specifically binds to the secretory substance may be selected from at least one of the group consisting of a first antibody, a first aptamer, and a molecularly imprinted polymer, and may include a plurality of types of first antibodies.

[0022] The cell evaluation apparatus according to the present technology may further include a second capturing unit for capturing cells, and the image acquisition unit and the second capturing unit may be sequentially provided in the cell evaluation apparatus according to the present technology.

[0023] In the cell evaluation apparatus according to the present technology, the first capturing unit may be provided around the second capturing unit.

[0024] In the cell evaluation device according to the present technology, the second capture unit includes a molecule that nonspecifically or specifically binds to the cell.

[0025] In the cell evaluation apparatus according to the present technology, a protection unit is provided in a region where the second capture unit does not include the molecule that nonspecifically or specifically binds to the cell.

[0026] The molecule that non-specifically or specifically binds to the cell may be at least one selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine, and may include a plurality of types of second antibodies.

[0027] In the cell evaluation apparatus according to the present technology, a photodegradable connector may be included between the second capturing unit and the image acquiring unit.

[0028] In addition, the present technology provides a cell evaluation device, comprising:

[0029] a first capturing unit, capturing secreted substances secreted by cells;

[0030] an image acquisition unit that acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to the secretory substance; and

[0031] an analyzing unit, analyzing the acquired image, wherein

[0032] The image acquisition unit and the first capture unit are arranged in sequence, and

[0033] The image acquisition unit acquires position information of cells and secreted substances.

[0034] In the cell evaluation apparatus according to the present technology, the analysis unit quantifies the secretory substance based on the intensity of the light emitted from the luminescent substance bound to the secretory substance.

[0035] Based on the brightness information of the observation light of the cells and / or the light emitted from the luminescent substance bound to the secreted substance, the analyzing unit can analyze the cells.

[0036] The cell evaluation apparatus according to the present technology may further include an irradiation unit that projects light onto the cells.

[0037] The cell evaluation apparatus according to the present technology may further include a display unit that displays an image.

[0038] The display unit may include: a well image display unit based on a cell image obtained by imaging observation light of the cell and a chemiluminescent image obtained by imaging light emitted from a luminescent substance bound to the secretory substance; and an analysis data display unit and an analysis graph display unit based on analysis by the analysis unit.

[0039] In addition, the present technology provides a cell evaluation system, comprising:

[0040] a first capturing unit, capturing secreted substances secreted by cells;

[0041] an image acquisition unit that acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to the secretory substance; and

[0042] an analyzing unit, analyzing the acquired image, wherein

[0043] The image acquisition unit and the first capture unit are arranged in sequence, and

[0044] The image acquisition unit acquires position information of cells and secreted substances.

[0045] In addition, the present technology provides a cell evaluation device, comprising:

[0046] The first antibody captures secreted substances secreted by the cells;

[0047] Secondary antibody, capture cells; and

[0048] A CMOS image sensor acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to a secreted substance, wherein

[0049] The CMOS image sensor, the first antibody and the second antibody are set in sequence.

[0050] The first antibody is disposed around the second antibody, and

[0051] CMOS image sensors acquire location information of cells and secreted substances, and

[0052] Provided is a cell evaluation device, comprising:

[0053] The first antibody captures secreted substances secreted by the cells;

[0054] secondary antibody, capture cells;

[0055] a CMOS image sensor that acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to the secreted substance; and

[0056] an analyzing unit, analyzing the acquired image, wherein

[0057] The CMOS image sensor, the first antibody, and the second antibody are arranged in this order.

[0058] The first antibody is disposed around the second antibody, and

[0059] CMOS image sensors acquire location information of cells and secreted substances.

[0060] Effects of the Invention

[0061] According to the present technology, cells and secretory substances produced from cells can be accurately and precisely evaluated. Note that the effects described here are not necessarily limitative and may be any effects described in the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 : is a block diagram showing a configuration example of a cell evaluation apparatus according to the first embodiment to which the present technology is applied.

[0063] Figure 2 is a diagram showing a configuration example of a cell evaluation apparatus according to the first embodiment to which the present technology is applied.

[0064] Figure 3 is a diagram showing a configuration example of a cell evaluation apparatus according to the first embodiment to which the present technology is applied.

[0065] Figure 4 is a cross-sectional view showing a configuration example of a CMOS image sensor (solid-state imaging element) including an antibody layer.

[0066] Figure 5 is a diagram showing a configuration example of an analysis unit.

[0067] Figure 6 is a diagram showing a configuration example of a display unit.

[0068] Figure 7 is a block diagram showing a configuration example of a cell evaluation apparatus according to a second embodiment to which the present technology is applied.

[0069] Figure 8 is a diagram showing an example of patterning of the first capturing unit and the second capturing unit.

[0070] Fig. 9 is a diagram showing an example of patterning of the first capturing unit and the second capturing unit.

[0071] Fig.10 is a diagram showing an example of patterning of the first capturing unit and the second capturing unit. DETAILED DESCRIPTION

[0072] Hereinafter, preferred embodiments for implementing the present technology will be described. The embodiments described below illustrate representative embodiments of the present technology, and the scope of the present technology is not narrowly interpreted by the embodiments.

[0073] Note that the description will be given in the following order.

[0074] 1. Overview of the cell evaluation device according to the present technology

[0075] 2. First Embodiment (Example 1 of Cell Evaluation Apparatus)

[0076] 2-1. Cell evaluation equipment

[0077] 2-2. First capture unit

[0078] 2-3. Image acquisition unit

[0079] 2-4. Analysis unit

[0080] 2-5. Irradiation unit

[0081] 2-6. Display unit

[0082] 3. Second Embodiment (Example 2 of Cell Evaluation Apparatus)

[0083] 3-1. Cell evaluation equipment

[0084] 3-2. First Capture Unit and Second Capture Unit, and Patterning of the First Capture Unit and the Second Capture Unit

[0085] 3-3. Image acquisition unit, analysis unit, irradiation unit, and display unit

[0086] 4. Third Embodiment (Example of Cell Evaluation System)

[0087] 4-1. Cell evaluation system

[0088] 4-2. First Capturing Unit, Second Capturing Unit, Image Acquisition Unit, Analysis Unit, Illumination Unit, and Display Unit

[0089] <1. Overview of cell evaluation apparatus according to the present technology>

[0090] The overview of the cell evaluation apparatus according to the present technology will be described.

[0091] As methods for acquiring secretory substances produced from cells, methods called the ELISA method and the ELISPOT method, and specialized systems (for example, an intracellular cytokine measurement system using flow cytometry, etc.) have been widely used.

[0092] The ELISA method is to culture a large number of cells in a well, collect only the secreted substances after removing the cells, and measure the secretion amount. In this case, it is impossible to find out what percentage of cells secrete the secreted substances. Therefore, in order to focus on each cell, research and development have been conducted, in which cells are cultured in smaller wells with one to several cells as units, and the secretion amount produced by each cell is calculated.

[0093] Meanwhile, in the ELISPOT method, a large number of cells are cultured in a well, secretory substances produced by cells are captured with a membrane placed at the bottom of the well, the cells are removed, and then reacted with the secretory substances attached to the membrane to color the secretory substances. The number of cells that secrete the secretory substances can be calculated by counting the number of colored spots observed under a microscope. However, by any of these methods, it may be difficult to find cells that produce secretory substances, or it may be impossible to classify each cell in which multiple cells coexist. Thereafter, a method has been proposed in which each single cell is placed in a small well for measurement. However, since cells are placed in wells respectively, it is impossible to observe interactions when multiple cells coexist, etc. In addition, in the ELISPOT method, since cells must be removed before measurement as described above, it is impossible to take out only specific cells after ELISPOT and then perform post-analysis (genetic analysis, etc.).

[0094] The present technology was achieved in view of the above circumstances, and even in an environment where multiple cells coexist, it is possible to find out which cell secretes how much. As a result, it is possible to understand the interaction that occurs between different cells such as cancer cells and immune cells, which is difficult as an application.

[0095] Examples of cells observed by the cell evaluation device according to the present technology include cancer cells, immune cells, etc. Specifically, immune cells include T cells, NK cells, macrophages, etc. In addition, the cells also include tissues in which a single or multiple cells having a certain function are assembled. In addition, examples of secreted substances produced from cells observed by the cell evaluation device according to the present technology include IFN-γ, interleukin, chemokine, etc.

[0096] Since the present technology can at least add the quantification of the secretory substances of each cell and the imaging of the cells to the function of ELISPOT (counting the number of secretory cells), the cell type and the state of the secretory substances that have been produced can be identified. For example, in the co-culture of cancer cells and immune cells, it is not currently found whether the cancer cells or immune cells have issued IFNγ, and it is expected that only the number of living cells that have produced IFNγ can be counted. For example, the problem of producing IFNγ during apoptosis can be solved. Since the present technology has at least the technology based on ELISPOT method and ELISA method, the number of required blood samples can be reduced. Since cells can be collected in principle, only cells that have produced secretory substances can be reanalyzed. Even in an environment where multiple cells coexist, it is also possible to evaluate which cells secrete how much. That is, as described above, it is difficult to understand the interaction between different cells, such as cancer cells and immune cells.

[0097] Hereinafter, in order to specifically describe the cell evaluation device and the cell evaluation system according to the present technology, the first embodiment of the present technology (Example 1 of the cell evaluation device), the second embodiment of the present technology (Example 2 of the cell evaluation device) and the third embodiment of the present technology (Example of the cell evaluation system) will be described.

[0098] <2. First Embodiment (Example 1 of Cell Evaluation Apparatus)>

[0099] [2-1. Cell evaluation equipment]

[0100] A cell evaluation device according to a first embodiment of the present technology (Example 1 of a cell evaluation device) is a cell evaluation device comprising the following units: a first capturing unit that captures secretory substances secreted by cells; and an image acquisition unit that acquires an image obtained by imaging observation light of the cells and light emitted from a luminescent substance bound to the secretory substances, wherein the image acquisition unit and the first capturing unit are arranged in sequence, and the image acquisition unit acquires positional information of the cells and the secretory substances.

[0101] Here, the image acquisition unit and the first capture unit are disposed in sequence, which means that the image acquisition unit and the first capture unit may be disposed continuously, or the image acquisition unit and the first capture unit may be disposed via an optional material layer (which may be a material film, the same below) or a material unit. In addition, the image acquisition unit and the first capture unit are disposed continuously, which means that the first capture unit is disposed directly above the image acquisition unit. Furthermore, the first capture unit being disposed directly above the image acquisition unit means, for example, that the first capture unit is disposed at a position 150 μm or less from the image acquisition unit, preferably at a position 10 μm or less from the image acquisition unit.

[0102] The cell evaluation device according to the first embodiment of the present technology may further include at least one of an analysis unit, an irradiation unit, or a display unit. That is, the cell evaluation device according to the first embodiment of the present technology may further include an analysis unit, may further include an irradiation unit, may further include a display unit, may further include an analysis unit and an irradiation unit, may further include an analysis unit and a display unit, may further include an irradiation unit and a display unit, or may further include an analysis unit, an irradiation unit, and a display unit.

[0103] Reference Figures 1 to 3 A specific example of the cell evaluation apparatus according to the first embodiment of the present technology is described. Figure 1 is a block diagram illustrating an example of a cell evaluation apparatus according to the first embodiment of the present technology. Figure 2 and Figure 3 is a configuration diagram showing an example of a cell evaluation apparatus according to the first embodiment of the present technology.

[0104] First, see Figure 1 The cell evaluation device 1 includes at least an upper portion 101 of an image sensor, which includes a first capture unit 101; an image sensor 1001 (e.g., a CMOS image sensor) which is an image acquisition unit; and an analysis unit 1005. In addition, the cell evaluation device 1 also includes a driver 1002, an illumination control unit 1004, a display unit 1006, and an illumination unit 1007. Figure 1 As shown in FIG. 1 , the irradiation control unit 1004, the analysis unit 1005 and the display unit 1006 can be formed as one device 1003. Figure 1 Although not shown in the figure, the image sensor upper portion 101 may include a protection unit in addition to the first capturing unit 101. The irradiation control unit 1004 may control the wavelength selectivity (light color selectivity) and exposure time of the light emitted by the irradiation unit 1007.

[0105] In the cell evaluation device 1, the first capture unit 101-1 (for example, a capture unit including a first antibody for capturing a secreted substance) is arranged above the image sensor 1001 in a desired positional relationship. In order to identify cells, an observation light or irradiation unit 1007 of the cells is used to capture a cell image, and image analysis of the cell size and cell shape can be performed. The cell type and cell state (for example, whether the cell is alive or dead) can be accurately found. In addition, the irradiation unit may include a digital mirror device (DMD) or the like to control the irradiation position.

[0106] In addition, cells are cultured above the image sensor 1001, and secretory substances produced from the cells are captured by the first capture unit 101-1 (for example, a capture unit including a first antibody for capturing secretory substances). Thereafter, by further causing a reaction with a chemiluminescent substance or the like, weak light is generated, and the light can be obtained by the image sensor 1001 directly below the first capture unit 101-1. The chemiluminescent substance is, for example, an enzyme, and the enzyme reacts with the substrate to emit chemiluminescence.

[0107] The driver 1002 photoelectrically converts weak light into an electrical signal, and increases only the brightness of the sensor portion where the secretory substance has been captured. Therefore, the position where the secretory substance has been obtained above the image sensor 1001 can be drawn. Since the positional relationship between the first capture unit 101-1 (for example, a capture unit including a first antibody for capturing the secretory substance) and the cell can be pre-specified, the cell that has produced the secretory substance can also be identified. In addition, the analysis unit 1005 can also quantify the secretory substance by utilizing the brightness change depending on the concentration of the secretory substance. The image obtained from the image sensor 1001 is displayed on the display unit 1006 (for example, a monitor) by the analysis unit 1005.

[0108] Examples of measures for increasing sensitivity include increasing the density of the first antibody used to capture the secreted substance contained in the first capture unit 101-1, minimizing the distance between the image sensor 1001 and the first capture unit 101-1 (e.g., the antibody layer) (disposing the first capture unit 101-1 directly above the image sensor 1001), increasing the exposure time, and separating the background noise part and the signal part from each other through image processing. In addition, the acquisition of the cell image and the detection of the secreted substance can also be performed in real time above the image sensor 1001.

[0109] Next, see Figure 2 The cell evaluation device 2 includes an antibody layer (first capture unit) 201 containing an antibody (first antibody) 200, a CMOS image sensor 2001 as an image acquisition unit, a driver 2002, and a device 2003 including an analysis unit and a display unit. Figure 2, the CMOS image sensor 2001 is shown for three pixels.

[0110] The CMOS image sensor 2001 includes a protective layer 202, a semiconductor substrate 203, and a wiring layer 204 in order from the light incident side. Figure 2 Three photodiodes (PDs) in the semiconductor substrate 203 are formed. Figure 2 The antibody layer 201 is disposed directly above the protective layer 202. By disposing the antibody layer 201 just above the protective layer 202, the CMOS image sensor 2001 can effectively receive light emitted from the luminescent substance bound to the secreted substance captured by the antibody 200 included in the antibody layer 201, as well as observation light of the cells. The observation light of the cells refers to light obtained by projecting natural light or room light onto the cells, for example.

[0111] exist Figure 2 In the figure, four wells are shown ( Figure 2 In the cell evaluation device 2, there are two wells in the horizontal direction and two wells in the vertical direction), which means that four wells are provided just above one CMOS image sensor 2001 (including the protective layer 205 in the CMOS image sensor 2001) in the cell evaluation device 2. In each well, for example, an antibody 200 that captures a secretory substance is fixed on the collagen coating (protective unit), pretreated cells are captured, cells are stimulated and cultured, secretory substances are produced from the cells, and the antibody 200 captures the secretory substance.

[0112] In addition, see Figure 3 The cell evaluation device 3 includes an antibody layer (first capture unit) 301 containing the antibody 300, a CMOS image sensor 3001 as 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 Figure 3 In FIG. 3 , the CMOS image sensor 3001 is shown for three pixels.

[0113] The CMOS image sensor 3001 includes a protective layer 302, a semiconductor substrate 303, and a wiring layer 304 in order from the light incident side. Figure 3 Three photodiodes (PDs) in the semiconductor substrate 303 are formed. Figure 3 The antibody layer 301 is disposed directly above the protective layer 302. By disposing the antibody layer 301 just above the protective layer 302, the CMOS image sensor 3001 can effectively receive light emitted from the luminescent substance bound to the secreted substance captured by the antibody 300 included in the antibody layer 301, and light obtained by projecting light onto the cells by the irradiation unit 3004.

[0114] exist Figure 3 In the figure, four wells are shown (in Figure 3 In the cell evaluation device 3, there are four wells in the vertical direction (four wells in the vertical direction), which means that one well is set just above one CMOS image sensor 3001 (including the protective layer 305 in the CMOS image sensor 3001) in the cell evaluation device 3. That is, the cell evaluation device 3 requires at least four CMOS image sensors 3001. In each well, for example, an antibody 300 that captures a secretory substance is fixed on the collagen coating (protective unit), pretreated cells are captured, cells are stimulated and cultured, secretory substances are produced from the cells, and the antibody 300 captures the secretory substance.

[0115] [2-2. First Capture Unit]

[0116] 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 secretory substances secreted by cells. The first capture unit may include molecules that bind to the secretory substances non-specifically or specifically. The molecules that bind to the secretory substances non-specifically or specifically may be at least one selected from the group consisting of a first antibody, a first aptamer, and an imprinted polymer, and may include multiple types of first antibodies. A protective unit may be provided in an area where the first capture unit does not include molecules that bind to the secretory substances non-specifically or specifically. The protective unit may include, for example, a collagen coating as described above. In addition, the protective unit may include at least one type of hydrophilic polymer capable of inhibiting the adsorption of biopolymers. Examples of hydrophilic polymers include polyethylene glycol, polymers having phosphorylcholine groups in their side chains, polysaccharides, polypeptides, and the like.

[0117] [2-3. Image acquisition unit]

[0118] The cell evaluation device according to the first embodiment of the present technology includes an image acquisition unit. The image acquisition unit can be, for example, an image sensor, and the image sensor can be, for example, a complementary metal oxide semiconductor (CMOS) image sensor or a solid-state imaging element such as a charge coupled device (CCD). Hereinafter, a back-illuminated solid-state imaging element and a front-illuminated solid-state imaging element as examples of a CMOS image sensor will be described in detail.

[0119] (Back-illuminated solid-state imaging element)

[0120] Will see Figure 4 A back-illuminated solid-state imaging element is described as an example of a CMOS image sensor. Figure 4 is a cross-sectional view showing a configuration example of one pixel 20 of the back-illuminated solid-state imaging element 10 .

[0121] The pixel 20 includes an organic photoelectric conversion element 41 and photodiodes 36 and 37 in one pixel, each of which has a pn junction, stacked in the depth direction. In addition, on the back side ( Figure 4 A protective layer 44 is formed on the organic photoelectric conversion element 41 (the upper part of the organic photoelectric conversion element 41). Immediately above the protective layer 44, an antibody layer 50 containing an antibody 51 is formed. That is, the protective layer 44 and the antibody layer 50 are continuously provided. In this case, the thickness of the protective layer 44 can be as thin as, for example, 100 nm. The protective layer 44 provided continuously with the antibody layer 50 preferably has transparency so as to effectively bring light into the organic photoelectric conversion element 41 and the photodiodes 36 and 37. The protective layer 44 preferably contains, for example, SiO2. The pixel 20 includes a semiconductor substrate (silicon substrate) 35, and the photodiodes 36 and 37 are formed in the semiconductor substrate 35. A light receiving surface on which light is incident is formed on the back side ( Figure 4 The upper side of the semiconductor substrate 35 in the pixel 20 is formed with a circuit including a reading circuit and the like on the front side of the semiconductor substrate 35. That is, the pixel 20 includes 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 first conductive type, such as an n-type semiconductor substrate.

[0122] In the semiconductor substrate 35, an inorganic photoelectric conversion unit having two pn junctions, that is, a first photodiode 36 and a second photodiode 37 are formed to be stacked in the depth direction from the back side. In the semiconductor substrate 35, the first photodiode 36 is formed, and the second photodiode 37 is formed in the depth direction from the back side (downward in the figure). Figure 4 In FIG. 1 , the first photodiode 36 is used for blue (B), and the second photodiode 37 is used for red (R).

[0123] Over the back surface of the semiconductor substrate 35, in the region where the first photodiode 36 and the second photodiode 37 are formed, an organic photoelectric conversion element 41 for the first color is provided, in which the first electrode (lower electrode) 33, the first buffer layer 47, the photoelectric conversion layer 32, the second buffer layer 46, and the second electrode (upper electrode) 31 are stacked in this order. Figure 4 In the example of the back-illuminated solid-state imaging element 10 shown in FIG, the organic photoelectric conversion element 41 is used for green (G). The second electrode (upper electrode) 31 and the first electrode (lower electrode) 33 can be respectively composed of a transparent conductive film such as an indium tin oxide film or an indium zinc oxide film.

[0124] As a combination of colors, Figure 4In the example of the back-illuminated solid-state imaging element 10 shown in FIG, the organic photoelectric conversion element 41 is used for green, the first photodiode 36 is used for blue, and the second photodiode 37 is used for red. However, another color combination may be used. For example, the organic photoelectric conversion element 41 may be used for red or blue, and the first photodiode 36 and the second photodiode 37 may be used for 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.

[0125] In addition, without using the first photodiode 36 and the second photodiode 37, three photoelectric conversion elements, that is, the organic photoelectric conversion element 41B for blue, the organic photoelectric conversion element 41G for green, and the organic photoelectric conversion element 41R for red can be applied to the solid-state imaging element (back-illuminated solid-state imaging element and front-illuminated solid-state imaging element) according to the second embodiment of the present technology. As the organic photoelectric conversion element 41B that performs photoelectric conversion using blue wavelength light, an organic photoelectric conversion material containing coumarin-based dyes, tris-8-hydroxyquinoline (A1q3) (tris-8-hydryxyquinoline A1 (A1q3)), meracyanine-based dyes, etc. can be used. As the organic photoelectric conversion element 41G that performs photoelectric conversion using green wavelength light, an organic photoelectric conversion material containing, for example, rhodamine-based dyes, meracyanine-based dyes, quinacridone, etc. can be used. As the organic photoelectric conversion element 41R that performs photoelectric conversion using red wavelength light, an organic photoelectric conversion material containing a phthalocyanine-based dye can be used.

[0126] In addition, in addition to the organic photoelectric conversion element 41B for blue, the organic photoelectric conversion element 41G for green, and the organic photoelectric conversion element 41R for red, the organic photoelectric conversion element 41UV for ultraviolet light and / or the organic photoelectric conversion element 41IR for infrared light can be applied to the solid-state imaging element (back-illuminated solid-state imaging element and front-illuminated solid-state imaging element) according to the second embodiment of the present technology. By providing the photoelectric conversion element 41UV for ultraviolet light and / or the photoelectric conversion element 41IR for infrared light, light having a wavelength outside the visible light region can be detected.

[0127] The light emitted from the luminescent substance by binding to the secreted substance captured by the antibody 51 (first antibody) contained in the antibody layer 50, the observation light of the cell, and the projection light obtained by projecting light onto the cell by the irradiation unit may each have any wavelength band, and imaging may be performed using at least one of the organic photoelectric conversion element 41, the first photodiode 36, or the second photodiode 37 according to the color (wavelength band) of these types of light. For example, in the solid-state imaging element 10, if the light emitted from the luminescent substance by binding to the secreted substance is green light, imaging may be performed using the organic photoelectric conversion element 41. If the light emitted from the luminescent substance by binding to the secreted substance is blue light, imaging may be performed using the first photodiode 36. If the light emitted from the luminescent substance by binding to the secreted substance is red light, imaging may be performed using the first photodiode 36. In addition, for example, if the observation light of the cell or the light obtained by projecting light onto the cell by the irradiation unit is achromatic light (e.g., white light, gray light, etc.), imaging may be performed using all of the organic photoelectric conversion element 41, the first photodiode 36, and the second photodiode 37.

[0128] In the organic photoelectric conversion element 41 , a first electrode (lower electrode) 33 is formed, and an insulating film 34 for dielectrically isolating the first electrode (lower electrode) 33 is formed below the first electrode (lower electrode) 33 in the figure.

[0129] 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. For example, in order to suppress a short circuit with Si, the wiring 39 and the wiring connected to the second electrode (upper electrode) 31 may be formed of SiO2, a tungsten (W) plug having a SiN insulating layer at its periphery, a semiconductor layer by ion implantation, or the like. Figure 2 In the example of the back-illuminated solid-state imaging element shown in , since the signal charge is a hole, when the wiring 39 is formed with a semiconductor layer by ion implantation, the wiring 39 is a p-type semiconductor layer. As the wiring connected to the second electrode (upper electrode) 31, since the second electrode (upper electrode) 31 extracts electrons, an n-type semiconductor layer can be used.

[0130] In this example, an n-type region 38 for charge accumulation is formed on the front face side of the semiconductor substrate 35. This n-type region 38 serves as a floating diffusion portion of the photoelectric conversion element 41.

[0131] As the insulating film 34 on the back side of the semiconductor substrate 35, a film having a negative fixed charge can be used. Examples of the film having a negative fixed charge include a hafnium oxide film. That is, the insulating film 34 can be formed to have a three-layer structure obtained by sequentially forming a silicon oxide film, a hafnium oxide film, and a silicon oxide film from the back side.

[0132] The wiring layer 45 is formed on the front surface side of the semiconductor substrate 35 ( Figure 4 Note that, although not shown, a color filter, a transparent filter, an ND filter, a white filter, a gray filter, or the like may be formed in the back-illuminated solid-state imaging element 10, or an on-chip lens may be formed.

[0133] (Front-illuminated solid-state imaging element)

[0134] The CMOS image sensor is not limited to the above-mentioned back-illuminated solid-state imaging element, and may be a front-illuminated solid-state imaging element. A front-illuminated solid-state imaging element will be described.

[0135] The example of the front-illuminated solid-state imaging element is different from the above-mentioned back-illuminated solid-state imaging element 10 only in that the wiring layer 92 formed under the semiconductor substrate 35 in the back-illuminated solid-state imaging element is formed between the organic photoelectric conversion element 41 in the front-illuminated solid-state imaging element and the semiconductor substrate 35. Other points may be similar to those of the above-mentioned back-illuminated solid-state imaging element 10, and description thereof is omitted here.

[0136] [2-4. Analysis unit]

[0137] As described above, the cell evaluation device according to the first embodiment of the present technology may include an analysis unit. Based on the intensity of light emitted from the luminescent substance bound to the secretory substance, the analysis unit can quantify the secretory substance. In addition, based on the observation light of the cell or the light (brightness information) obtained by projecting light onto the cell by the irradiation unit, the analysis unit can analyze the cell. In the analysis unit, the projected image of the cell can be converted into a higher resolution image to analyze the cell by holographic technology, super-resolution technology, etc.

[0138] Reference Figure 5 Describes the specific configuration of the analysis unit. Figure 5 An example of the configuration of the analysis unit is shown. Specifically, Figure 5 (a) shows antibody arrangement information 70a, Figure 5 (b) shows a chemiluminescent image, and Figure 5 (c) shows a cell image.

[0139] See also Figure 5 (a), the antibody arrangement information 70a indicates antibodies 71a to 76a. Each of the antibodies 71a to 76a includes, for example, a first antibody that has captured a secretory substance. In the case of a cell evaluation device according to a first embodiment of the present technology described later, each of the antibodies 71a to 76a includes, for example, a first antibody that has captured a secretory substance and a second antibody that has captured a cell.

[0140] See also Figure 5(b), the chemiluminescent image 70b has separate chemiluminescent images 71b to 76b. The chemiluminescent images 71b to 76b correspond to the antibodies 71a to 76a, respectively. The chemiluminescent intensity can be calculated from the brightness information of the antibody arrangement area of ​​the chemiluminescent images 71b to 76b, and the amount of the secreted substance can be further calculated. Specifically, in the quantification of the secreted substance, the intensity of the chemiluminescence is obtained from the brightness information of each pixel obtained from the image sensor, and the concentration of the secreted substance is calculated from the intensity and information of the calibration curve previously obtained as data.

[0141] See also Figure 5 In (c), the cell image 70c indicates the individual cell images 71c to 76c. Figure 5 In (c), cell images 71c to 76c are cell images with a magnification of 1 relative to the cell (having the actual size of the cell), but may be cell images obtained by enlarging the cell or by reducing the cell as required. Because cell images 71c to 76c correspond to antibodies 71a to 76a and chemiluminescent images 71b to 76b, respectively, the concentration of the secreted substance of each of the cell images 71c to 76c can be uniquely determined. Then, object segmentation is performed based on the brightness information of the observation light of the cell or cell images 71c to 76c captured by the irradiation unit. The size, position, and shape of each of the cell images 71c to 76c can be analyzed. The internal state of each cell can be analyzed from the dynamic range within each cell, etc. In addition, the total number of cells included in one well can be calculated. For example, the size of the cell represented as cell image 71c corresponds to arrow P7. Note that in the case of a cell evaluation device according to the second embodiment of the present technology described later, since each cell of cell images 71c to 76c is a cell captured by the second antibody, the cell position information becomes clearer.

[0142] [2-5. Irradiation unit]

[0143] As described above, the cell evaluation device according to the first embodiment of the present technology may include an irradiation unit. The irradiation unit is a device for determining a cell image, and as described above, the wavelength selectivity (photochromatic selectivity) and exposure time of the light emitted by the irradiation unit can be controlled using an irradiation control unit. Therefore, a clearer cell image can be obtained using the irradiation unit. In addition, only specific cells can be released by cleaving the photodegradable linker using a specific wavelength emitted.

[0144] [2-6. Display unit]

[0145] The cell evaluation device according to the first embodiment of the present technology may include a display unit. The display unit includes: a well image display unit based on a cell image obtained by imaging light (brightness information) obtained by projecting light onto the cell by an irradiation unit or by imaging light emitted from a luminescent substance bound to a secretory substance; and an analysis data display unit and an analysis graph display unit based on the analysis of the analysis unit.

[0146] Will see Figure 6 Describes the specific configuration of the display unit. Figure 6 An example of a configuration of a display unit is shown. Specifically, Figure 6 (b) shows the Figure 6 The well image display unit of (a) shows a chemiluminescent image 80a-1 and a cell image 80a-2. Figure 6 (c) shows an analysis data display unit. Figure 6 (d) shows an analysis graph display unit.

[0147] See also Figure 6 In (a) and (b), in the well display unit 80b, images in which the chemiluminescent images 81a-1 to 86a-1 of the chemiluminescent image 80a-1 are associated with the cell images 81a-2 to 86a-2 are respectively shown. Figure 6 (b) shows information about cells in the well 1A included in a total of six wells 1A to 3A and 2A to 3B. Figure 6 (b), chemiluminescent image 81b-1 corresponds to cell image 81b-2, chemiluminescent image 82b-1 corresponds to cell image 82b-2, chemiluminescent image 83b-1 corresponds to cell image 83b-2, chemiluminescent image 84b-1 corresponds to cell image 84b-2, chemiluminescent image 85b-1 corresponds to cell image 85b-2, and chemiluminescent image 86b-1 corresponds to cell image 86b-2. In these images, Figure 6 Among the six cells shown in (b), the cell image 81a-2 (cell image 81b-2) having the cell size indicated by arrow P8 has the largest cell size and the largest chemiluminescence amount. In this case, the cell image 81a-2 is Figure 6 (a) and Figure 6 In contrast, the cells of the cell image 83a-2 (cell image 83b-2) have a minimum cell size to a level where the cells are not displayed, and have a chemiluminescence amount of almost zero. In this case, the cell image 83a-2 is shown in white as a chemiluminescent image. Figure 6 (a) and Figure 6 The chemiluminescent image in (b) is shown in black.

[0148] See also Figure 6 (c), the analysis data display unit 80c indicates that the total number of cells in well 1A is 10 4 , and also indicates the cell diameter (μm) of each cell (cell number), its secretion amount (pg), etc.

[0149] See also Figure 6 (d), the analysis graph display unit 80d indicates a graph in which the vertical axis represents the secretion amount (pg) and the horizontal axis represents the cell size (cell diameter) (μm). Figure 6 According to the correlation between the secretion amount (pg) and the cell size (cell diameter) (μm), the number of cells to be evaluated can be determined as 10 4 100 of them. Figure 6 In (d), cells indicated by black circles among the 100 cells correspond to, for example, the cells of the above-mentioned cell image 81a-2 (cell image 81b-2).

[0150] <3. Second Embodiment (Example 2 of Cell Evaluation Apparatus)>

[0151] [3-1. Cell evaluation equipment]

[0152] The cell evaluation device according to the second embodiment of the present technology (Example 2 of the cell evaluation device) is a cell evaluation device including the following units: a first capture unit that captures secreted substances secreted by cells; a second capture unit that captures cells; and an image acquisition unit that acquires an image obtained by imaging observation light of the cells and light emitted from a luminescent substance combined with the secreted substance, wherein the image acquisition unit and the first capture unit are arranged in sequence, and furthermore, the image acquisition unit and the second capture unit are arranged in sequence, and the image acquisition unit acquires position information of the cells and the secreted substance. That is, the cell evaluation device according to the second embodiment of the present technology (Example 2 of the cell evaluation device) has a configuration in which the second capture unit is added to the cell evaluation device according to the first embodiment of the present technology (Example 1 of the cell evaluation device). Except for the configuration of the second capture unit, the configuration of the cell evaluation device according to the second embodiment of the present technology (Example 2 of the cell evaluation device) can be the same as the configuration of the cell evaluation device according to the first embodiment of the present technology (Example 1 of the cell evaluation device). Here, the phrase that the image acquisition unit and the first capture unit are arranged in sequence means that the image acquisition unit and the first capture unit can be arranged continuously, or the image acquisition unit and the first capture unit can be arranged via an optional material layer (material film) or material unit. In addition, the image acquisition unit and the first capture unit are continuously arranged, which means that the first capture unit is arranged just above the image acquisition unit. The first capture unit being arranged just above the image acquisition unit means, for example, that the first capture unit is arranged at a position 150 μm or less from the image acquisition unit, preferably at a position 10 μm or less from the image acquisition unit.

[0153] In addition, the image acquisition unit and the second capture unit are arranged in sequence, which means that the image acquisition unit and the second capture unit can be arranged continuously, or the image acquisition unit and the second capture unit can be arranged via an optional material layer (material film) or material unit. In addition, the image acquisition unit and the second capture unit are arranged continuously, which means that the second capture unit is arranged directly above the image acquisition unit. The second capture unit is arranged directly above the image acquisition unit, which means that, for example, the first capture unit is arranged at a position 150μm or less from the image acquisition unit, preferably at a position 10μm or less from the image acquisition unit.

[0154] The cell evaluation device according to the second embodiment of the present technology may further include at least one of an analysis unit, an irradiation unit, or a display unit. That is, the cell evaluation device according to the second embodiment of the present technology may further include an analysis unit, may further include an irradiation unit, may further include a display unit, may further include an analysis unit and an irradiation unit, may further include an analysis unit and a display unit, may further include an irradiation unit and a display unit, or may further include an analysis unit, an irradiation unit, and a display unit.

[0155] See also Figure 7 The cell evaluation device 7 includes at least an upper portion 101 of an image sensor, which includes 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. In addition, the cell evaluation device 1 also includes a driver 1002, an irradiation control unit 1004, a display unit 1006, and an irradiation unit 1007. Figure 1 As shown in FIG. 1 , the irradiation control unit 1004, the analysis unit 1005 and the display unit 1006 can be formed as one device 1003. Figure 7 Although not shown in the figure, the upper portion 101 of the image sensor may include a protection unit in addition to the first capturing unit 101-1 and the second capturing unit 101-2.

[0156] In the cell evaluation device 7, a first capture unit 101-1 (for example, a capture unit including a first antibody for capturing a secretory substance) and a second capture unit 101-2 (for example, a capture unit including a second antibody for capturing cells) are set above the image sensor 1001 in a desired positional relationship. In order to identify the cells captured by the second capture unit 101-2, the cell image is captured using the observation light or irradiation unit 1007 of the cell, and the cell size and cell shape can be image analyzed. The cell type and cell state (for example, whether the cell is alive or dead) can be accurately found. The second capture unit 101-2 captures the cell. Therefore, after culturing the cells, for example, by optical linker technology, etc., only the specific capture antibody (second antibody) that has reacted with the secretory substance is lysed, and the cells can be collected. In this case, it is preferred to include a photodegradable linker between the second capture unit 101-2 and the image sensor 1001. The inclusion of a photodegradable linker only allows specific cells to be released. That is, the cell evaluation device 7 is capable of releasing cells by irradiating them with light after analyzing the cells, and taking out only desired cells for post-analysis (gene analysis, etc.).

[0157] In addition, cells are cultured above the image sensor 1001, and secretory substances produced from the cells are captured by the first capture unit 101-1 (for example, a capture unit including a first antibody for capturing secretory substances). Thereafter, by further causing a reaction with a chemiluminescent substance or the like, weak light is generated, and the light can be obtained by the image sensor 1001 directly below the first capture unit 101-1. The chemiluminescent substance is, for example, an enzyme, and the enzyme reacts with the substrate to emit chemiluminescence.

[0158] The driver 1002 photoelectrically converts weak light into an electrical signal, and increases the brightness of only the sensor portion where the secretion substance has been captured. Therefore, the position where the secretion substance has been obtained above the image sensor 1001 can be drawn. Since the positional relationship between the first capture unit 101-1 (e.g., a capture unit including a first antibody for capturing the secretion substance) and the second capture unit 101-2 (e.g., a capture unit including a second antibody for capturing cells) can be pre-specified, the cells that have emitted the secretion substance can also be identified. In addition, the analysis unit 1005 can also quantify the secretion substance by utilizing the brightness change depending on the concentration of the secretion substance. The image obtained from the image sensor 1001 is displayed on the display unit 1006 (e.g., a monitor) by the analysis unit 1005.

[0159] Examples of measures for increasing sensitivity include increasing the density of the first antibody used to capture the secreted substance contained in the first capture unit 101-1, minimizing the distance between the image sensor 1001 and the first capture unit 101-1 (e.g., the antibody layer) (disposing the first capture unit 101-1 directly above the image sensor 1001), increasing the exposure time, and separating the background noise part and the signal part from each other through image processing. In addition, the acquisition of the cell image and the detection of the secreted substance can also be performed in real time above the image sensor 1001.

[0160] [3-2. First Capture Unit and Second Capture Unit, and Patterning of the First Capture Unit and the Second Capture Unit]

[0161] The cell evaluation device according to 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 secreted substances secreted by cells. The first capture unit is as described in the part of the cell evaluation device according to the first embodiment of the present technology (Example 1 of the cell evaluation device), so a detailed description thereof is omitted here.

[0162] The second capture unit captures the cell. The second capture unit may include molecules that bind to the cell non-specifically or specifically. The molecules that bind to the cell non-specifically or specifically may be at least one selected from the group consisting of a second antibody, a second aptamer, a supramolecule, and oleylamine, and may include multiple types of second antibodies. In the area where the second capture unit does not include molecules that bind to the cell non-specifically or specifically, a protection unit may be provided. The protection unit may include, for example, a collagen coating. In addition, the protection unit may include at least one type of hydrophilic polymer that can inhibit the adsorption of biopolymers. Examples of hydrophilic polymers include polyethylene glycol, polymers having a phosphorylcholine group in their side chains, polysaccharides, polypeptides, and the like.

[0163] Will refer to Figures 8 to 10 The patterning of the first capture unit and the second capture unit is described. Figure 8 (a), in a size of 50 μm, the first capture unit 402a is disposed around the second capture unit 401a, and the protection unit 403a is disposed between the first capture unit 402a and the second capture unit 401a. Figure 8 (b) shows 16 patterns, each pattern includes Figure 8 (a) shows a first capture unit and a second capture unit. Figure 8 The second capture unit 401a shown in (a) corresponds to Figure 8 The second capture unit 401b shown in (b) of Figure 8 The first capture unit 402a shown in (a) corresponds to Figure 8 The second capture unit 402b shown in (b) of FIG. Figure 8 The protection unit 403a shown in (a) corresponds to Figure 8 The second protection unit 403b is shown in (b).

[0164] Note that the cell evaluation apparatus according to the first embodiment (Example 1 of the cell evaluation apparatus) of the present technology does not include the second capturing unit 401a, and thus the first capturing unit 402a is provided in part of the second capturing unit 401a.

[0165] See also Fig. 9 (a), in a size of 50 μm, the first capture units 502a to 505a are arranged around the second capture unit 501a, and the protection unit 506a is arranged between the first capture units 502a to 505a and the second capture unit 501a and in the first capture units 502a to 505a. The first capture units 502a and 503a include the same material (e.g., a first antibody), the first capture units 503a and 504a include the same material (e.g., a first aptamer), and two types of materials are used. Regarding the first capture units 502a to 505a, in Fig. 9 In (a), the first capture units 502a and 503a both have a vertical rectangular shape (the longitudinal direction is Fig. 9 The first capture units 504a and 505a both have a transverse rectangular shape (the longitudinal direction is Fig. 9 In the lateral direction of (a), the first capture units comprising different materials are adjacent to each other. Fig. 9 (b) shows 16 patterns, each pattern includes Fig. 9 (a) shows a first capture unit and a second capture unit.

[0166] Fig. 9The second capture unit 501a shown in (a) corresponds to Fig. 9 The second capture unit 501b shown in (b) of Fig. 9 The first capture unit 502a shown in (a) corresponds to Fig. 9 The second capture unit 502b shown in (b) of FIG. Fig. 9 The first capture unit 503a shown in (a) corresponds to Fig. 9 The second capture unit 503b shown in (b) of FIG. Fig. 9 The first capture unit 504a shown in (a) corresponds to Fig. 9 The second capture unit 504b shown in (b) of FIG. Fig. 9 The first capture unit 505a shown in (a) corresponds to Fig. 9 The second capture unit 505b shown in (b), and Fig. 9 The protection unit 506a shown in (a) corresponds to Fig. 9 The protection unit 506b is shown in (b).

[0167] Note that the cell evaluation apparatus according to the first embodiment of the present technology (Example 1 of the cell evaluation apparatus) does not include the second capture unit 501a, and thus at least one of the first capture units 502a to 505a is provided in part of the second capture unit 501a.

[0168] See also Fig.10 (a), in a size of 50 μm, the first capture units 602a to 609a are disposed around the second capture unit 601a, and the protection unit 610a is disposed between the first capture units 602a to 609a and the second capture unit 601a and in the first capture units 602a to 609a. The first capture units 602a to 609a contain materials different from each other (e.g., eight first antibodies or eight first aptamers), and eight materials are used. Fig.10 In (a), eight square first capturing units 602a to 609a are provided. Fig.10 (b) shows 16 patterns, each pattern includes Fig.10 (a) shows a first capture unit and a second capture unit.

[0169] Fig.10 The second capture unit 601A shown in (a) corresponds to Fig.10 The second capture unit 601(b) shown in (b) of FIG. Fig.10 The first capture unit 602a shown in (a) corresponds to Fig.10 The second capture unit 602b shown in (b) of FIG. Fig.10The first capture unit 603a shown in (a) corresponds to Fig.10 The second capture unit 603b shown in b, Fig.10 The first capture unit 604a shown in (a) corresponds to Fig.10 The second capture unit 604b shown in (b) of FIG. Fig.10 The first capture unit 605a shown in (a) corresponds to Fig. 9 The second capture unit 605b shown in (b) of FIG. Fig.10 The first capture unit 606a shown in (a) corresponds to Fig.10 The second capture unit 606b shown in (b) of FIG. Fig.10 The first capture unit 607a shown in (a) corresponds to Fig.10 The second capture unit 607b shown in (b) of FIG. Fig.10 The first capture unit 608a shown in (a) corresponds to Fig.10 The second capture unit 608b shown in (b) of FIG. Fig.10 The first capture unit 609a shown in (a) corresponds to Fig.10 The second capture unit 609b shown in (b), and Fig.10 The protection unit 610a shown in (a) corresponds to Fig.10 The protection unit 610b shown in (b) of . Note that the cell evaluation device according to the first embodiment (Example 1 of the cell evaluation device) of the present technology does not include the second capture unit 601a, and therefore at least one of the first capture units 602a to 609a is set in a part of the second capture unit 601a.

[0170] [3-3. Image Acquisition Unit, Analysis Unit, Illumination Unit, and Display Unit]

[0171] The image acquisition unit included in the cell evaluation device according to the second embodiment (Example 2 of the cell evaluation device) of the present technology is as described in the part of the cell evaluation device according to the first embodiment (Example 1 of the cell evaluation device) of the present technology, and therefore a detailed description thereof is omitted here. In addition, the analysis unit, the irradiation unit, and the display unit that may be included in the cell evaluation device according to the second embodiment (Example 2 of the cell evaluation device) of the present technology are also as described in the part of the cell evaluation device according to the first embodiment (Example 1 of the cell evaluation device) of the present technology, and therefore a detailed description thereof is omitted here.

[0172] <4. Third Embodiment (Example of Cell Evaluation System)>

[0173] [4-1. Cell evaluation system]

[0174] A cell evaluation system according to a third embodiment (an example of a cell evaluation system) of the present technology is a cell evaluation system comprising the following units: a first capturing unit that captures secretory substances secreted by cells; an image acquisition unit that acquires an image obtained by imaging observation light of the cells and light emitted from a luminescent substance bound to the secretory substances; and an analysis unit that analyzes the obtained image, wherein the image acquisition unit and the first capturing unit are arranged in sequence, and the image acquisition unit acquires positional information of the cells and the secretory substances.

[0175] In addition, a cell evaluation system according to a third embodiment of the present technology (an example of a cell evaluation system) may be a cell evaluation system comprising: a first capturing unit that captures secretory substances secreted by cells; an image acquisition unit that acquires an image obtained by imaging observation light of the cells and light emitted from a luminescent substance bound to the secretory substances; and an analysis unit that analyzes the obtained image, wherein the image acquisition unit and the first capturing unit are arranged in sequence, and the image acquisition unit acquires positional information of the cells and the secretory substances.

[0176] The cell evaluation system according to the third embodiment of the present technology may further include at least one of an irradiation unit or a display unit. That is, the cell evaluation system according to the third embodiment of the present technology may further include an irradiation unit, may further include a display unit, or may further include an irradiation unit and a display unit.

[0177] Note that for the cell evaluation system according to the third embodiment of the present technology, in addition to the above contents, the contents described in the parts of the cell evaluation devices according to 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 according to the third embodiment of the present technology.

[0178] [4-2. First Capturing Unit, Second Capturing Unit, Image Acquisition Unit, Analysis Unit, Illumination Unit, and Display Unit]

[0179] The image acquisition unit and the analysis unit included in the cell evaluation system according to the third embodiment (cell evaluation system) of the present technology are as described in the part of the cell evaluation device according to the first embodiment (Example 1 of the cell evaluation device) of the present technology, and therefore their detailed description is omitted here. In addition, the illumination unit and the display unit that may be included in the cell evaluation system according to the third embodiment (cell evaluation system) of the present technology are also as described in the part of the cell evaluation device according to the first embodiment (Example 1 of the cell evaluation device) of the present technology, and therefore their detailed description is omitted here. In addition, the second capture unit that may be included in the cell evaluation system according to the third embodiment (cell evaluation system) of the present technology is as described in the part of the cell evaluation device according to the second embodiment (Example 2 of the cell evaluation device) of the present technology, and therefore its detailed description is omitted here.

[0180] Note that embodiments of the present technology are not limited to the above-described embodiments, and various modifications may be made thereto without departing from the gist of the present technology.

[0181] Furthermore, the effects described here are merely examples, and the effects of the present technology are not limited thereto and may include other effects.

[0182] In addition, the present technology may also have the following configurations [1] to

[68] .

[0183] [1] A cell evaluation device comprising:

[0184] a first capturing unit that captures secreted substances secreted by cells; and

[0185] an image acquisition unit that acquires an image obtained by imaging observation light of the cell and light emitted from the luminescent substance bound to the secretory substance, wherein

[0186] The image acquisition unit and the first capture unit are arranged in sequence, and

[0187] The image acquisition unit acquires position information of cells and secreted substances.

[0188] [2] The cell evaluation device according to [1], wherein the image acquisition unit is a CMOS image sensor.

[0189] [3] The cell evaluation apparatus according to [1] or [2], wherein the image acquisition unit acquires an image obtained by imaging chemiluminescence emitted by causing the luminescent substance as an enzyme to react with a substrate.

[0190] [4] The cell evaluation device according to any one of [1] to [3], wherein the first capture unit includes a molecule that non-specifically or specifically binds to the secretory substance.

[0191] [5] The cell evaluation apparatus according to [4], wherein a protection unit is provided in a region of the first capture unit that does not include the molecule that non-specifically or specifically binds to the secretory substance.

[0192] [6] The cell evaluation device according to [4] or [5], wherein the molecule that non-specifically or specifically binds to the secretory substance is at least one selected from the group consisting of a first antibody, a first aptamer, and a molecularly imprinted polymer.

[0193] [7] The cell evaluation device according to [6], comprising a plurality of types of first antibodies.

[0194] [8] The cell evaluation apparatus according to any one of [1] to [7], further comprising a second capturing unit that captures cells, wherein the image acquisition unit and the second capturing unit are arranged in sequence.

[0195] [9] The cell evaluation device according to [8], wherein the first capture unit is arranged around the second capture unit.

[0196]

[10] The cell evaluation device according to [8] or [9], wherein a photodegradable connector is included between the second capturing unit and the image acquiring unit.

[0197]

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

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

[0198]

[12] The cell evaluation apparatus according to

[11] , wherein a protection unit is provided in a region of the second capture unit that does not include the molecule that non-specifically or specifically binds to the cell.

[0199]

[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.

[0200]

[14] The cell evaluation device according to

[13] , which comprises a plurality of types of secondary antibodies.

[0201]

[15] A cell evaluation device comprising:

[0202] a first capturing unit, capturing secreted substances secreted by cells;

[0203] an image acquisition unit that acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to the secretory substance; and

[0204] an analyzing unit, analyzing the acquired image, wherein

[0205] The image acquisition unit and the first capture unit are arranged in sequence, and

[0206] The image acquisition unit acquires position information of cells and secreted substances.

[0207]

[16] The cell evaluation apparatus according to

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

[0208]

[17] The cell evaluation device according to

[15] or

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

[0209]

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

[15] to

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

[0210]

[19] The cell evaluation apparatus according to any one of

[15] to

[18] , wherein the image acquisition unit acquires an image obtained by imaging chemiluminescence emitted by causing the luminescent substance as an enzyme to react with a substrate.

[0211]

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

[15] to

[19] , wherein the first capture unit includes a molecule that non-specifically or specifically binds to the secretory substance.

[0212]

[21] The cell evaluation device according to

[20] , wherein a protection unit is provided in a region of the first capture unit that does not include the molecule that non-specifically or specifically binds to the secretory substance.

[0213]

[22] The cell evaluation device according to

[20] or

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

[0214]

[23] The cell evaluation device according to

[22] , which comprises a plurality of types of first antibodies.

[0215]

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

[15] to

[23] further includes a second capturing unit that captures cells, wherein the image acquisition unit and the second capturing unit are arranged in sequence.

[0216]

[25] The cell evaluation device according to

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

[0217]

[26] The cell evaluation device according to

[24] or

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

[0218]

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

[24] to

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

[0219]

[28] The cell evaluation device according to

[27] , wherein a protection unit is provided in a region of the second capture unit that does not include molecules that nonspecifically or specifically bind to cells.

[0220]

[29] The cell evaluation device according to

[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.

[0221]

[30] The cell evaluation device according to

[29] , which comprises a plurality of types of secondary antibodies.

[0222]

[31] The cell evaluation apparatus according to any one of [1] to

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

[0223]

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

[31] , further comprising a display unit that displays an image.

[0224]

[33] A cell evaluation device according to

[32] , wherein the display unit includes: a well image display unit based on a cell image obtained by imaging observation light of the cells and a chemiluminescent image obtained by imaging light emitted from a luminescent substance bound to a secretory substance; and an analysis data display unit and an analysis graph display unit based on analysis by the analysis unit.

[0225]

[34] A cell evaluation system, comprising:

[0226] a first capturing unit that captures secreted substances secreted by cells; and

[0227] an image acquisition unit that acquires an image obtained by imaging observation light of the cell and light emitted from the luminescent substance bound to the secretory substance, wherein

[0228] The image acquisition unit and the first capture unit are arranged in sequence, and

[0229] The image acquisition unit acquires position information of cells and secreted substances.

[0230]

[35] The cell evaluation system according to

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

[0231]

[36] The cell evaluation system according to

[34] or

[35] , wherein the image acquisition unit acquires an image obtained by imaging chemiluminescence emitted by causing the luminescent substance as an enzyme to react with a substrate.

[0232]

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

[34] to

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

[0233]

[38] The cell evaluation system according to

[37] , wherein a protection unit is provided in a region of the first capture unit that does not include the molecule that non-specifically or specifically binds to the secretory substance.

[0234]

[39] The cell evaluation system according to

[37] or

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

[0235]

[40] The cell evaluation system according to

[39] , which comprises a plurality of types of first antibodies.

[0236]

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

[34] to

[40] further includes a second capturing unit for capturing cells, wherein the image acquisition unit and the second capturing unit are arranged in sequence.

[0237]

[42] The cell evaluation system according to

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

[0238]

[43] The cell evaluation system according to

[41] or

[42] , wherein a photodegradable connector is included between the second capturing unit and the image acquiring unit.

[0239]

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

[41] to

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

[0240]

[45] The cell evaluation system according to

[44] , wherein a protection unit is provided in a region of the second capture unit that does not include the molecule that non-specifically or specifically binds to the cell.

[0241]

[46] The cell evaluation system according to

[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.

[0242]

[47] The cell evaluation system according to

[46] , which comprises a plurality of types of secondary antibodies.

[0243]

[48] ​​A cell evaluation system, comprising:

[0244] a first capturing unit, capturing secreted substances secreted by cells;

[0245] an image acquisition unit that acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to the secretory substance; and

[0246] an analyzing unit, analyzing the acquired image, wherein

[0247] The image acquisition unit and the first capture unit are arranged in sequence, and

[0248] The image acquisition unit acquires position information of cells and secreted substances.

[0249]

[49] The cell evaluation system according to

[48] , 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.

[0250]

[50] A cell evaluation system according to

[48] or

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

[0251]

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

[48] to

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

[0252]

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

[48] to

[51] , wherein the image acquisition unit acquires an image obtained by imaging chemiluminescence emitted by causing the luminescent substance as an enzyme to react with a substrate.

[0253]

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

[48] to

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

[0254]

[54] The cell evaluation system according to

[53] , wherein a protection unit is provided in a region of the first capture unit that does not include the molecule that non-specifically or specifically binds to the secretory substance.

[0255]

[55] The cell evaluation system according to

[53] or

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

[0256]

[56] The cell evaluation system according to

[55] , comprising a plurality of types of first antibodies.

[0257]

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

[48] to

[56] further includes a second capturing unit for capturing cells, wherein the image acquisition unit and the second capturing unit are arranged in sequence.

[0258]

[58] The cell evaluation system according to

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

[0259]

[59] The cell evaluation system according to

[57] or

[58] , wherein a photodegradable connector is included between the second capturing unit and the image acquiring unit.

[0260]

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

[57] to

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

[0261]

[61] The cell evaluation system according to

[60] , wherein a protection unit is provided in a region of the second capture unit that does not include the molecule that non-specifically or specifically binds to the cell.

[0262]

[62] The cell evaluation system according to

[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.

[0263]

[63] The cell evaluation system according to

[62] , which comprises a plurality of types of secondary antibodies.

[0264]

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

[34] to

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

[0265]

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

[34] to

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

[0266]

[66] A cell evaluation system according to

[65] , wherein the display unit comprises: the display unit comprises: a well image display unit based on a cell image obtained by imaging the observation light of the cell and a chemiluminescent image obtained by imaging the light emitted from the luminescent substance bound to the secretory substance; and an analysis data display unit and an analysis graph display unit based on the analysis of the analysis unit.

[0267]

[67] A cell evaluation device comprising:

[0268] The first antibody captures secreted substances secreted by the cells;

[0269] Secondary antibody, capture cells; and

[0270] A CMOS image sensor acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to a secreted substance, wherein

[0271] The CMOS image sensor, the first antibody and the second antibody are set in sequence.

[0272] The first antibody is disposed around the second antibody, and

[0273] CMOS image sensors acquire location information of cells and secreted substances.

[0274]

[68] A cell evaluation device comprising:

[0275] The first antibody captures secreted substances secreted by the cells;

[0276] secondary antibody, capture cells;

[0277] a CMOS image sensor that acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to the secreted substance; and

[0278] an analyzing unit, analyzing the acquired image, wherein

[0279] The CMOS image sensor, the first antibody and the second antibody are set in sequence.

[0280] The first antibody is disposed around the second antibody, and

[0281] CMOS image sensors acquire location information of cells and secreted substances.

[0282]

[69] A cell evaluation system, comprising:

[0283] The first antibody captures secreted substances secreted by the cells;

[0284] Secondary antibody, capture cells; and

[0285] A CMOS image sensor acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to a secreted substance, wherein

[0286] The CMOS image sensor, the first antibody and the second antibody are set in sequence.

[0287] The first antibody is disposed around the second antibody, and

[0288] CMOS image sensors acquire location information of cells and secreted substances.

[0289]

[70] A cell evaluation system, comprising:

[0290] The first antibody captures secreted substances secreted by the cells;

[0291] secondary antibody, capture cells;

[0292] a CMOS image sensor that acquires an image obtained by imaging observation light of the cell and light emitted from a luminescent substance bound to the secreted substance; and

[0293] an analyzing unit, analyzing the acquired image, wherein

[0294] The CMOS image sensor, the first antibody and the second antibody are set in sequence.

[0295] The first antibody is disposed around the second antibody, and

[0296] CMOS image sensors acquire location information of cells and secreted substances.

[0297] Reference numerals list

[0298] 1 Cell evaluation equipment

[0299] 101-1 First Capture Unit

[0300] 101-2 Second Capture Unit

[0301] 1001 Image acquisition unit (image sensor)

[0302] 1005 Analysis Unit

[0303] 1006 Display Unit

Claims

1. A cell evaluation device, comprising: a first capturing unit, capturing secreted substances secreted by cells; a second capture unit, capturing the cell, and an image acquisition unit that acquires an image obtained by imaging the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance, wherein The first capturing unit is arranged around the second capturing unit, The image acquisition unit and the first capture unit are arranged in sequence, and The image acquisition unit acquires position information of the cell and the secreted substance, The cell evaluation device uniquely determines the concentration of the secretory substance of each cell based on the correspondence between a cell image obtained by imaging the observation light of the cell and a chemiluminescent image obtained by imaging the light emitted from the luminescent substance bound to the secretory substance.

2. The cell evaluation device according to claim 1, wherein The image acquisition unit is a CMOS image sensor.

3. The cell evaluation device according to claim 1, wherein The image acquisition unit acquires an image obtained by imaging chemiluminescence emitted by causing the luminescent substance as an enzyme to react with a substrate.

4. The cell evaluation device according to claim 1, wherein The first capture unit comprises a molecule that binds non-specifically or specifically to the secreted substance.

5. The cell evaluation device according to claim 4, wherein A protection unit is arranged in a region of the first capture unit that does not include the molecule that non-specifically or specifically binds to the secretory substance.

6. The cell evaluation device according to claim 4, wherein: The molecule that non-specifically or specifically binds to the secretory 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 claim 6, wherein: The first antibodies are of various types.

8. The cell evaluation device according to claim 1, wherein The second capture unit comprises a molecule that binds non-specifically or specifically to the cell.

9. The cell evaluation device according to claim 8, wherein A protection unit is disposed in an area of ​​the second capture unit that does not include the molecule that non-specifically or specifically binds to the cell.

10. The cell evaluation device according to claim 8, 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.

11. The cell evaluation device according to claim 10, wherein The second antibodies are of various types.

12. A cell evaluation device comprising: a first capturing unit, capturing secreted substances secreted by cells; a second capture unit, capturing the cell, an image acquisition unit that acquires an image obtained by imaging the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance; and an analyzing unit, analyzing the acquired image, wherein The first capturing unit is arranged around the second capturing unit, The image acquisition unit and the first capture unit are arranged in sequence, and The image acquisition unit acquires position information of the cell and the secreted substance, The cell evaluation device uniquely determines the concentration of the secretory substance of each cell based on the correspondence between a cell image obtained by imaging the observation light of the cell and a chemiluminescent image obtained by imaging the light emitted from the luminescent substance bound to the secretory substance.

13. The cell evaluation device according to claim 12, wherein The analyzing unit quantifies the secretion substance based on the intensity of the light emitted from the luminescent substance bound to the secretion substance.

14. The cell evaluation device according to claim 12, wherein The analyzing unit analyzes the cell based on the observation light of the cell and / or brightness information of the light emitted from the luminescent substance bound to the secretory substance. 15 . The cell evaluation apparatus according to claim 12 , further comprising an irradiation unit that projects light onto the cells. 16 . The cell evaluation apparatus according to claim 12 , further comprising a display unit that displays the image.

17. The cell evaluation device according to claim 16, wherein The display unit includes: a well image display unit based on a cell image obtained by imaging observation light of the cell and a chemiluminescent image obtained by imaging light emitted from a luminescent substance bound to the secretory substance; and an analysis data display unit and an analysis graph display unit based on analysis by the analysis unit.

18. A cell evaluation system, comprising: a first capturing unit, capturing secreted substances secreted by cells; a second capture unit, capturing the cell, an image acquisition unit that acquires an image obtained by imaging the observation light of the cell and the light emitted from the luminescent substance bound to the secretory substance; and an analyzing unit, analyzing the acquired image, wherein The first capturing unit is arranged around the second capturing unit, The image acquisition unit and the first capture unit are arranged in sequence, and The image acquisition unit acquires position information of the cell and the secreted substance, The cell evaluation system uniquely determines the concentration of the secretory substance of each cell based on the correspondence between a cell image obtained by imaging the observation light of the cell and a chemiluminescent image obtained by imaging the light emitted from the luminescent substance bound to the secretory substance.

Images