Information processing method, and information processing system

Abstract

To provide a method for improving the calculation accuracy of the amount or concentration of a target substance.SOLUTION: An information processing method detects a target substance. The information processing device includes an image acquisition step for acquiring an image including a plurality of individual separation sections that can include at least either one of the target substance and a target capture substance for capturing the target substance as a subject, a detection step for detecting the existence / nonexistence of the target capture substance in each of the individual separation sections in the image, and a calculation step for calculating the amount or concentration of the target substance on the basis of information on the number of the individual separation sections with the target capture substance detected and the number of sections with the target substance detected among the individual separation sections with the target capture substance detected, and information on the number of the individual separation sections with no target capture substance detected and the number of sections with the target substance detected among the individual separation sections with no target capture substance detected.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present disclosure relates to an information processing method and an information processing system.

Background Art

[0002] Quickly and accurately detecting a target substance in a sample is important in fields such as medical diagnosis and academic research. For example, in the medical field, the target substance is detected or its concentration is measured from the amount of signal emitted by a reporter in an assay of a target capture substance (including antibodies, etc.) that captures the target substance. In recent years, a highly sensitive measurement method has been developed as disclosed in Patent Document 1, in which such a detection reagent is fed into individual separation compartments, the individual separation compartments are isolated with a hydrophobic solvent, and the reaction is detected with a fluorescence microscope or the like.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] In an antibody-antigen reaction, the antigen and the antibody are constantly in a state of binding and dissociation, and the target capture substance and the target substance are not necessarily bound. Therefore, the dissociated target substance may be filled in an individual separation compartment without the target capture substance. However, in Patent Document 1, only the individual separation compartments in which the target capture substance is present are targeted for measurement, so the number of target substances to be measured is underestimated. The present disclosure provides a method for improving the calculation accuracy of the concentration of the target substance in response to such problems.

Means for Solving the Problems

[0005] The present disclosure is an information processing method for detecting a target substance, including an image acquisition step of acquiring an image including a plurality of individual separation compartments that may contain at least one of the target substance and a target capture substance that captures the target substance as a subject, a detection step of detecting the presence or absence of the target capture substance in each of the individual separation compartments in the image, and a calculation step of calculating the amount or concentration of the target substance based on information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected, and information regarding the number of the individual separation compartments in which the target capture substance is not detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected.

[0006] Another aspect of the present disclosure includes a placement unit for placing an aggregate including a plurality of individual separation compartments, an imaging unit for imaging an image including the plurality of individual separation compartments as a subject, and an information processing unit. The information processing unit includes an image acquisition unit for acquiring an image including a plurality of individual separation compartments that may contain at least one of the target substance and a target capture substance that captures the target substance as a subject, a detection unit for detecting the presence or absence of the capture substance in each of the individual separation compartments in the image, and a calculation unit for calculating the amount or concentration of the target substance based on information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected, and information regarding the number of the individual separation compartments in which the target capture substance is not detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected, and is a calculation system.

[0007] Yet another aspect of the present disclosure is an information processing system including an information processing unit that processes an image including a plurality of individually separated compartments as a subject. The information processing unit includes: an image acquisition unit that acquires an image including a plurality of individually separated compartments that may contain at least one of a target substance and a target capture substance that captures the target substance as a subject; a detection unit that detects the presence or absence of the capture substance in each of the individually separated compartments in the image; and a calculation unit that calculates the amount or concentration of the target substance based on information regarding the number of the individually separated compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individually separated compartments in which the target capture substance is detected, and information regarding the number of the individually separated compartments in which the target capture substance is not detected and the number of the compartments in which the target substance is detected among the individually separated compartments in which the target capture substance is not detected.

Advantages of the Invention

[0008] According to the present disclosure, the calculation accuracy of the concentration of the target substance can be improved.

Brief Description of the Drawings

[0009]

Figure 1

Figure 2

Figure 3

Figure 4A

Figure 4B

Figure 4C

Figure 4D

Figure 5

Figure 6

Embodiments for Carrying Out the Invention

[0010] Hereinafter, embodiments of the present disclosure will be exemplarily described in detail with reference to the drawings. However, the components described in this embodiment are merely examples, and the technical scope of the present disclosure is determined by the claims and is not limited by the following individual embodiments.

[0011] FIG. 1 shows an information processing method according to this embodiment. The information processing method of this embodiment is an information processing method for detecting a target substance using individual separation compartments, and includes an image acquisition step, a detection step, and a calculation step. The individual separation compartments may include at least one of a target substance and a target capture substance that captures the target substance.

[0012] The image acquisition step 1001 acquires an image including a plurality of individual separation compartments that may contain a target substance as a subject. The detection step 1002 detects the positions of the individual separation compartments and the presence or absence of the target capture substance from the image obtained in the image acquisition step 1001. The calculation step 1003 calculates the concentration of the target substance based on information regarding the number of individual separation compartments in which the target capture substance is detected and the number of compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected, and information regarding the number of individual separation compartments in which the target capture substance is not detected and the number of compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected.

[0013] As will be described in detail later, by calculating the concentration of the target substance using information regarding the number of individual separation compartments in which the target capture substance is not detected, it is possible to calculate a more accurate amount (number) or concentration, particularly when detecting a target substance with a low concentration.

[0014] <Individual Separation Compartment> The individual separation compartments that may contain the target substance in this embodiment will be described.

[0015] In this embodiment, an individual separation compartment refers to a compartment that is independently separated. The volume of the compartment is preferably minute, and preferably ranges from 0.1 fL to 1000 fL. Preferred examples of the individual separation compartment include droplets and wells of a well plate. An object containing a plurality of individual separation compartments may be referred to as an aggregate. For example, a well plate is an aggregate, and a liquid containing a plurality of droplets (which may include its container) is an aggregate.

[0016] As the droplets, water-in-oil emulsions (W / O emulsions) are preferably used. Such droplets can be prepared by a pumping method of an emulsion film.

[0017] An example of a well of the well plate in this embodiment is shown in FIG. 2. Well 204 is a recess for accommodating a liquid, and is separated from each other by a partition wall 203. Well 204 can have a lower substrate 201 as its bottom surface, and the shape of the region surrounded by the bottom surface and side surfaces of well 204 may be, for example, a cylindrical shape or a prismatic shape. When the shape of well 204 is a cylindrical shape with a circular bottom, the diameter of the bottom surface of well 204 is, for example, 0.5 μm or more and 12 μm or less, and the depth of well 204 is preferably 0.5 μm or more and 12 μm or less. Further preferably, the diameter of the bottom surface of well 204 is 1 μm or more and 9 μm or less, and the depth of well 204 is 1 μm or more and 9 μm or less. The upper substrate 202 preferably faces the opening of well 204 and the upper surface of the partition wall 203 with a space 205 therebetween. Space 205 serves as a flow path through which various liquids can flow, and the various liquids can flow from an injection port portion (not shown) to a discharge port portion respectively. As the well plate, a commercially available one such as a Simoa disk may be used, or it may be fabricated.

[0018] By filling a liquid containing a target substance into individual separation compartments, the sample appears to be concentrated, enabling the detection of the target substance without an amplification step and shortening the time until the signal saturates. The volume per compartment of the individual separation compartments can be made sufficiently small and set such that the amount of the target substance contained in one compartment is one molecule or less. By counting the number of compartments in which a signal is obtained, a digital assay for calculating the concentration of the target substance in the liquid becomes possible.

[0019] <Target substance> The target substance in the present embodiment is not particularly limited, and examples thereof include proteins, nucleic acids, lipids, sugars, low-molecular compounds, enzymes, receptors, antibodies, antigens, cytokines, hormones, and membrane proteins. The target substance in the present embodiment may contain at least any one of nucleic acids, antibodies, antigens, enzymes, and enzyme substrates.

[0020] The individual separation compartments in the present embodiment can contain a detection reagent, and the liquid containing the target substance can contain the detection reagent. The detection reagent preferably exhibits a signal by interacting with the target substance. Known substances can be used for the detection reagent without limitation. For example, when the target substance is a nucleic acid (hereinafter sometimes referred to as a target nucleic acid) and the CRISPR / Cas system is used, an effector protein, a crRNA that binds to the target nucleic acid, and a reporter molecule can be used as the detection reagent. That is, the crRNA binds to the target nucleic acid, activates the effector protein, and the activated effector protein modifies the reporter molecule to generate fluorescence as a signal. When using a target nucleic acid, it is not limited to the above system, and various probes that generate a signal by the presence of the target nucleic acid can be used. Here, generating a signal means generating fluorescence, a magnetic field, etc., and the generated fluorescence, magnetic field, etc. can be detected as a signal using appropriate detection means. A probe whose generated signal changes according to the amount or concentration of the target nucleic acid can be used.

[0021] Examples of detection reagents when the target substance in this embodiment is a protein include substances containing antibodies, modified antibodies, antigens, enzymes, and enzyme substrate aptamers, and these can be labeled with fluorescent dyes or generate signals by using appropriate enzymes. When the target substance is an enzyme, an enzyme substrate can be used as the detection reagent. The enzyme substrate may be labeled with a fluorescent dye and generate fluorescence, which is a signal, by being modified by the substrate.

[0022] In this embodiment, an effective digital assay can be achieved by setting the target substance per individual separation compartment to be one molecule or less. Alternatively, when using a target capture substance, the same effect can be obtained and a digital assay can be achieved by setting the number of target substances per target capture substance to be one molecule or less.

[0023] <Target capture substance> The target capture substance in this embodiment is not particularly limited as long as it can capture the target substance in this embodiment. Examples of the target capture substance in this embodiment include particles provided with a ligand. The particles may have a ligand in order to capture the target substance. The ligand may contain at least one of nucleic acid, antibody, antigen, enzyme, and enzyme substrate. Examples of the particles include polymer resin (such as styrene resin, acrylic resin) particles, silica particles, agarose carrier resin particles, metal particles, latex particles, magnetic particles, etc. By using the target capture substance, it becomes possible to separate the target substance by centrifugation or using magnetism. Preferably, particles with a particle size of 1 μm or more and 10 μm or less can be used.

[0024] The method of filling (dispensing) the liquid containing the target substance into the individual separation compartments in this embodiment will be described.

[0025] In this embodiment, when a well plate is used as the aggregate and wells are used as individual separation compartments, the description will be made again with reference to FIG. 2. The aggregate 200 (here, a well plate) is left under reduced pressure to degas the space 205. Specifically, it is preferable to leave the well plate in a desiccator under a reduced pressure of 0.1 atm for a predetermined time. By performing degassing, the air in the well 204 is removed, and the reaction solution can be efficiently filled into the well 204. The degassing time is not particularly limited and can be arbitrarily set. The method of filling the reaction solution is not limited to the method by degassing. Next, a hydrophobic solvent is fed into the space 205 for sealing. That is, the reaction solution present in the space 205 above the well 204 is replaced with a hydrophobic solvent. As the hydrophobic solvent, for example, fluorinated oil, saturated aliphatic hydrocarbon, unsaturated aliphatic hydrocarbon, aromatic hydrocarbon, silicone oil, etc. can be used. Examples of fluorinated oil include Fluorinert, Asahi Klin AE-3000 (manufactured by AGC), and Fomblin (manufactured by Solvay). Examples of saturated hydrocarbons include Isopar (manufactured by ExxonMobil) and mineral oil.

[0026] In this embodiment, when droplets are used as individual separation compartments, the liquid can be prepared by a pumping method of an SPG emulsion film, with the liquid as the dispersion layer and the continuous layer as a fatty hydrocarbon solvent containing a surfactant.

[0027] The signal generation in this embodiment will be described. The individual separation compartment filled with the liquid containing the target substance appropriately generates a signal in order to generate a signal.

[0028] For example, in the example using the above CRISPR / Cas system, it is incubated at 37°C. By this incubation, the trans-cleavage reaction of CRISPR-Cas proceeds, and fluorescence derived from the fluorescent substance possessed by the reporter molecule is generated. In the case where the target substance is an enzyme, signal generation can be performed by proceeding with the reaction at the optimum temperature of the enzyme.

[0029] <Image acquisition step> Next, the image acquisition step will be described. The image acquisition unit acquires an image including a plurality of individually separated compartments as a subject. The image includes all the information contained in the image, for example, all kinds of information such as luminance, color, and shading for each coordinate, and the image acquisition unit can acquire the necessary information among these all kinds of information. The image may be extracted. The image may be the whole of the aggregate, that is, the whole of the well plate, or the whole of the liquid including the droplets contained in the container as the subject, or a part of the aggregate, that is, a part of the well plate, or a part of the liquid including the droplets as the subject. The image preferably includes at least either a bright-field image or an image capable of acquiring information on a signal indicating the presence of the target substance. An example of an image capable of acquiring information on a signal indicating the presence of the target substance is a fluorescence image. The image acquisition unit is not limited as long as it can acquire an image, and an imaging device such as a microscope or a CCD camera integrated with the information processing system may be the image acquisition unit, or it may be a means connected to these to acquire the information of the captured image.

[0030] The image includes particles and wells in the individually separated compartments. In the image acquisition unit, the image does not necessarily need to be acquired in one shot, and the shooting may be performed multiple times and combined into one image. When the individually separated compartment is a well, one image can include the whole image of one aggregate, but it is not necessarily the case that one image corresponds to one aggregate, and one image may include a plurality of aggregates, or only a part of the aggregate. For example, in a commercially available microscope for imaging well plates, it is known that about 6×103 wells are imaged in one shot, this is repeated 100 times, and these shots are concatenated to form an image of 6×105 wells.

[0031] An example of an image including an aggregate as a subject to be photographed in this embodiment is shown in FIG. 3. FIG. 3 is an image obtained by imaging a plurality of individually separated compartments detected by a CRISPR / Cas system with a nucleic acid as a target substance using a well plate with a fluorescence microscope. Particles were used as target capture substances. The left is a bright-field image, and the right is a fluorescence image. Particles can be confirmed in the bright-field image of FIG. 3. In the fluorescence image, fluorescence derived from the detection reagent can be confirmed. Also, in the well that is glowing in the center of the left end of the fluorescence image of FIG. 3, no particles can be confirmed in the bright field. That is, this is a case where the target substance dissociates from the particles and is accidentally confined in this individually separated compartment and reacts to emit fluorescence.

[0032] <Detection step> The information processing method in this embodiment has a detection step of detecting the positions of individually separated compartments and the positions of target capture substances. In the detection step, a position image of the individually separated compartments and a position image of the target capture substances are created. Either a bright-field image as shown in FIG. 4A or an image capable of acquiring information on signals indicating the presence of the target substance and the individually separated compartments is used. Thereby, a position image of the individually separated compartments in which the positions of each individually separated compartment as shown in FIG. 4B are detected and a position image of the individually separated compartments in which the target capture substances are detected, in which the positions of the individually separated compartments including the target capture substances as shown in FIG. 4D are detected, can be created. Here, the position image of the individually separated compartments in which the target capture substances are detected is, for example, an image obtained by extracting the wells containing particles.

[0033] Regarding the method for creating a position image of the individually separated compartments in this embodiment, the following methods can be considered. For example, when the aggregate is a well plate, since the wells, which are individually separated compartments, are regularly arranged, a method of creating a position image of the individually separated compartments in advance and adjusting the translation direction and angle by template matching or the like can be used. A method of emphasizing the outer periphery of the individually separated compartments using edge detection such as a Sobel filter and detecting circles by Hough transform or the like can also be considered.

[0034] In addition, a mask image obtained by extracting a target capture substance as shown in FIG. 4C is created, and by superimposing this on the position image of the individual separation compartments, a position image of the individual separation compartments in which the target capture substance is detected as shown in FIG. 4D can be obtained.

[0035] <Calculation step> The calculation step in the present embodiment calculates the luminance value of each individual separation compartment while distinguishing between the individual separation compartments in which the target capture substance is detected and the individual separation compartments in which the target capture substance is not detected. In the present embodiment, the calculation step can be performed based on the position image of the individual separation compartments obtained in the detection step and the position image of the individual separation compartments in which the target capture substance is detected. Note that the luminance value refers to the luminance in a bright-field image, and in an image capable of acquiring information on a signal indicating the presence of a target substance, it refers to the intensity of the signal indicating the presence of the target substance (for example, fluorescence intensity). The luminance value is a statistical value of the pixel values within the individual separation compartment, and can be any of an average value, a median value, a minimum value, and a maximum value.

[0036] The calculation step according to the present embodiment may include a step of performing negative and positive determinations based on the relationship between a predetermined threshold value and the luminance value of the individual separation compartments in which the target capture substance is detected, and the relationship between the predetermined threshold value and the luminance value of the individual separation compartments in which the target capture substance is not detected. Specifically, for the calculated luminance value (group) of the individual separation compartments in which the target capture substance is detected and the luminance value (group) of the individual separation compartments in which the target capture substance is not detected, a determination as to whether it is a negative individual separation compartment or a positive individual separation compartment is made using a predetermined threshold value for each. This determination is sometimes referred to as a negative and positive determination. Note that a positive individual separation compartment indicates a compartment containing a target substance, and a negative individual separation compartment indicates a compartment not containing a target substance. That is, it is classified into the following four types. · Positive individual separation compartments in which the target capture substance is detected · Negative individual separation compartments in which the target capture substance is detected · Positive individual separation compartments in which the target capture substance is not detected · Negative individual separation compartments in which the target capture substance is not detected

[0037] Whether to make it a positive individual separation section or a negative individual separation section can be determined, for example, by setting a threshold value for information on the intensity (signal intensity, luminance value) of a signal in an image from which information on a signal indicating the presence of a target substance can be obtained. For example, the threshold value may be automatically determined using the Otsu method or the like from the luminance values of each individual separation section, or the threshold value may be determined from the variation in the signal intensity based on an aggregate that does not contain the target substance. The threshold value may also be determined based on the signal intensity of an individual separation section that does not contain the target capture substance.

[0038] Despite the absence of the target capture substance, the reason for the increase in the luminance value of the individual separation section is considered to be that in the liquid, the target substance is released from the target capture substance at a certain rate and is accidentally isolated in an individual separation section without the target capture substance. The dissociation model can be expressed as shown in the following formula (1) in the case of an antibody-antigen reaction. Here, Ab is an antibody, Ag is an antigen, and AbAg indicates a state in which the antibody and the antigen are bound. Ab + Ag ⇔ AbAg ··· Formula (1)

[0039] As shown in the above formula (1), it can be seen that the antibody and the antigen bind and dissociate at a certain rate. Also, the rate of this binding and dissociation is determined by the following formula (2). Kd = [Ab][Ag] / [AbAg] ··· Formula (2)

[0040] Here, [Ab] is the concentration of the antibody, [Ag] is the concentration of the antigen, [AbAg] is the concentration at which the antibody and the antigen are bound, and Kd is the dissociation constant. The dissociation constant varies for each antibody, and from formula 2, the binding ratio depends on the concentrations of the antibody and the antigen.

[0041] This dissociated target substance may enter an individual separation section containing the target capture substance or may enter an individual separation section not containing the target capture substance. In the conventional method, only the individual separation section in which the target capture substance is present is the measurement target, but in this embodiment, even in an individual separation section not containing the target capture substance, it is the measurement target for calculating the concentration of the target substance.

[0042] Actually, to calculate the measured values, it is necessary to separate the individual separated compartments where the target capture substance was detected from those where the target capture substance was not detected. First, in the case of an individual separated compartment where the target capture substance was detected, the APR (Average Positive Rate), which indicates the ratio of positive compartments in the measurement target compartments, is calculated as shown in Equation (3). APR)b = BP / (BP + BN) ··· Equation (3)

[0043] Here, APRb is the APR of the individual separated compartment where the target capture substance was detected, BP is the number of positive individual separated compartments where the target capture substance was detected, and BN is the number of negative individual separated compartments where the target capture substance was detected.

[0044] Similarly, even in the case of an individual separated compartment where the target capture substance was not detected, the APR is calculated using Equation (4). APRw = WP / (WP + WN) ··· Equation (4)

[0045] Here, APRw is the APR of the individual separated compartment without the target capture substance, WP is the number of positive individual separated compartments where the target capture substance was not detected, and WN is the number of negative individual separated compartments where the target capture substance was not detected.

[0046] Next, the APR(P(k)) is calculated from the APRs of the individual separated compartments where the target capture substance was detected and those where the target capture substance was not detected, respectively. When the number of target substances contained in the sample is large, there is a possibility that two or more target substances are contained in one individual separated compartment. Therefore, there may be a case where the number of target substances does not match the number of individual separated compartments generating signals. For the above reasons, it is preferable to calculate the concentration of the target substance by calculation considering the Poisson distribution. In the Poisson distribution, when the average number of target substances per individual separated compartment is λ, the ratio P(k) of wells generating signals can be expressed by the following Equation (5). P(k / λ) = (λk / k!)e-1 (k = 0, 1, 2, ···) ··· Equation (5)

[0047] P(k) can be obtained from the number of individual separated compartments in which a signal is generated, and λ, which is the number of target substances, can be calculated.

[0048] When the average number of target substances in the individual separated compartments where the target capture substance is detected is λb, and the average number of target substances in the individual separated compartments where the target capture substance is not detected is λw, the concentration C of the target substance in the liquid can be calculated by Equation (6). C = λb × bead + λw × {(La - Lb) / Lw} ··· Equation (6)

[0049] Here, bead is the total number of target capture substances put into the liquid, La is the volume of the liquid, Lw is the volume of the individual separated compartment, and Lb is the total volume of the individual separated compartments where the target capture substance is detected.

[0050] By using the information processing method according to this embodiment, since the target substances dissociated from the target capture substances are also detection targets, it is possible to improve the measurement sensitivity and measurement accuracy. As a result, it is possible to improve the measurement stability in the measurement when the target substance is at a low concentration, and thus it is possible to expand the detection limit.

[0051] In addition, since the concentration of the dissociated target substance and the concentration of the bound target substance are known, if the dissociation constant of the antibody is known, it is possible to confirm whether the assay is successful using Equation 2.

[0052] In addition, the present disclosure can provide a program for executing the information processing method according to the above embodiment.

[0053] The information processing system and the calculation system according to this embodiment will be described with reference to FIG. 5.

[0054] The calculation device according to this embodiment includes a placement unit 103, an imaging unit 104, and an information processing unit 105, and the information processing unit 105 includes an image acquisition unit 1001, a detection unit 1002, and a calculation unit 1003.

[0055] The placement unit 103 places the assembly 200. The placement unit may have a fixing part adapted to the assembly in order to place the assembly 200. The imaging unit 104 captures an image including a plurality of individual separation compartments included in the assembly as subjects. The imaging unit is connected to the information processing unit 105. The calculation system in the present embodiment may further include a filling unit 101, a signal generation unit 102, and a display unit 109. The filling unit 101 may include a robotic arm, a pipette, a device for degassing, a pumping device, etc. for filling the individual separation compartments with a liquid containing a target substance. The signal generation unit includes an incubator, a stirrer, a permeation device, etc. so that the individual separation compartments filled with the liquid containing the target substance generate signals. The display unit includes a monitor of a personal computer. The calculation system according to the present embodiment may have each part in one device or may have them separately. For example, the information processing unit may be provided on the cloud, and information may be exchanged by communication means.

[0056] The hardware configuration of the information processing unit 105 in the present embodiment will be described with reference to FIG. 6.

[0057] The information processing unit 105 in the present embodiment has computer functions. For example, the information processing system may be integrally configured with a desktop PC (Personal Computer), a laptop PC, a tablet PC, a smartphone, etc. The information processing unit 105 may have a function of controlling the operations of the filling unit 101, the signal generation unit 102, and the imaging unit 104 according to a predetermined program.

[0058] In this embodiment, the information processing unit 105 includes a CPU (Central Processing Unit) 301, a RAM (Random Access Memory) 302, a ROM (Read Only Memory) 303, and an HDD (Hard Disk Drive) 304 in order to realize the functions of a computer that performs operations and storage. The information processing unit 105 also includes a communication I / F (interface) 306, a display device 307, and an input device 308. The CPU 301, the RAM 302, the ROM 303, the HDD 304, the communication I / F 306, the display device 307, and the input device 308 are interconnected via a bus 305. Note that the display device 307 and the input device 308 may be connected to the bus 305 via a drive device (not shown) for driving these devices.

[0059] In FIG. 6, each part constituting the information processing unit 105 is illustrated as an integrated device, but a part of these functions may be configured by an external device. For example, the display device 307 and the input device 308 may be external devices separate from the part that constitutes the functions of a computer including the CPU 301 and the like.

[0060] In this embodiment, the CPU 301 performs a predetermined operation according to a program stored in the RAM 302, the HDD 304, etc., and also has a function of controlling each part of the information processing unit 105. The RAM 302 is composed of a volatile storage medium and provides a temporary memory area necessary for the operation of the CPU 301. The ROM 303 is composed of a non-volatile storage medium and stores necessary information such as a program used for the operation of the information processing unit 105. The HDD 304 is a storage device composed of a non-volatile storage medium and stores information regarding the number and position of individual separation sections, fluorescence intensity, etc.

[0061] In this embodiment, the communication I / F 410 is a communication interface based on standards such as Wi-Fi (registered trademark) and 4G, and is a module for communicating with other devices. The display device 307 is a liquid crystal display, an OLED (Organic Light Emitting Diode) display, etc., and is used for displaying videos, still images, characters, etc. The input device 308 is a button, a touch panel, a keyboard, a pointing device, etc., and is used for a user to operate the information processing unit 105. The display device 307 and the input device 308 may be integrally formed as a touch panel.

[0062] Note that the hardware configuration shown in FIG. 6 is an example, and other devices may be added, or some devices may not be provided. Also, some devices may be replaced with other devices having similar functions. Further, some functions may be provided by other devices via a network, or the functions constituting this embodiment may be realized by being distributed among a plurality of devices. For example, the HDD 304 may be replaced with an SSD (Solid State Drive) using a semiconductor element such as a flash memory, or may be replaced with cloud storage.

[0063] In this embodiment, the CPU 301 realizes the functions of the image acquisition unit 1001, the exclusion region determination means 1002, and the calculation unit 1003 by loading and executing the programs stored in the ROM 303 etc. into the RAM 302. Also, the CPU 301 realizes the function of the display unit 109 by controlling the display device 307. Also, the CPU 301 realizes the function of the storage unit by controlling the HDD 304.

[0064] The information processing system according to this embodiment includes the above-described information processing unit.

[0065] The present invention can also be realized by supplying a program that realizes one or more functions of the above-described embodiments to a system or device via a network or a storage medium, and having one or more processors in a computer of the system or device read and execute the program. It can also be realized by a circuit (for example, ASIC) that realizes one or more functions.

[0066] Note that the above disclosure includes the following methods, programs, and configurations.

[0067] (Method 1) An information processing method for detecting a target substance, an image acquisition step of acquiring an image including a plurality of individual separation compartments that may include at least one of the target substance and a target capture substance that captures the target substance as a subject; a detection step of detecting the presence or absence of the target capture substance in each of the individual separation compartments in the image; information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected; and a calculation step of calculating the amount or concentration of the target substance based on information regarding the number of the individual separation compartments in which the target capture substance is not detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected.

[0068] (Method 2) The information processing method according to Method 1, wherein the individual separation compartment is a well or a droplet.

[0069] (Method 3) The information processing method according to Method 1 or 2, wherein the target capture substance includes particles.

[0070] (Method 4) The information processing method according to Method 3, wherein the particles include magnetic particles.

[0071] (Method 5) The information processing method according to method 3 or 4, wherein the particles have a particle size of 1 μm or more and 10 μm or less.

[0072] (Method 6) The information processing method according to any one of claims 1 to 5, wherein the target capture substance contains at least one of nucleic acid, antibody, antigen, enzyme, and enzyme substrate.

[0073] (Method 7) The information processing method according to any one of claims 1 to 6, wherein the target substance contains at least one of nucleic acid, antibody, antigen, enzyme, and enzyme substrate.

[0074] (Method 8) The information processing method according to any one of claims 1 to 7, wherein the image contains at least one of a bright-field image and a fluorescence image.

[0075] (Method 9) The information processing method according to any one of claims 1 to 8, wherein the volume of the individual separation compartment is 0.1 fL or more and 1000 fL or less.

[0076] (Method 10) The information processing method according to any one of claims 1 to 9, wherein the calculation step includes a step of performing negative and positive determinations based on the relationship between a predetermined threshold value and the luminance value of the individual separation compartment in which the target capture substance is detected, and the relationship between the predetermined threshold value and the luminance value of the individual separation compartment in which the target capture substance is not detected.

[0077] (Method 11) The information processing method according to any one of claims 1 to 10, wherein the calculation step includes a step of calculating the amount or concentration of the target substance using the average number of target substances in the individual separation compartment in which the target capture substance is detected and the average number of target substances in the individual separation compartment in which the target capture substance is not detected.

[0078] (Program) A program for executing the information processing method according to any one of claims 1 to 11.

[0079] (Configuration 1) A placement unit for placing an aggregate including a plurality of individual separation compartments; An imaging unit for imaging an image including a plurality of the individual separation compartments as subjects; An information processing unit, and includes: The information processing unit: An image acquisition unit that acquires an image including a plurality of individual separation compartments that may include at least one of a target substance and a target capture substance that captures the target substance as subjects; A detection unit that detects the presence or absence of the capture substance in each of the individual separation compartments in the image; Information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected; A calculation unit that calculates the amount or concentration of the target substance based on the number of the individual separation compartments in which the target capture substance is not detected and information regarding the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected; A calculation system having the above.

[0080] (Configuration 2) An information processing system including an information processing unit that processes an image including a plurality of individual separation compartments as subjects, The information processing unit: An image acquisition unit that acquires an image including a plurality of individual separation compartments that may include at least one of a target substance and a target capture substance that captures the target substance as subjects; A detection unit that detects the presence or absence of the capture substance in each of the individual separation compartments in the image; Information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected; A calculation unit that calculates the amount or concentration of the target substance based on the number of the individual separation compartments in which the target capture substance is not detected and information regarding the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected; An information processing system having

Explanation of symbols

[0081] 101 Filling unit 102 Signal generation unit 103 Mounting unit 104 Imaging unit 105 Information processing unit 109 Display unit

Claims

1. An information processing method for detecting a target substance, comprising: an image acquisition step of acquiring an image including a plurality of individual separation compartments that may contain at least one of the target substance and a target capture substance that captures the target substance as a subject; a detection step of detecting the presence or absence of the target capture substance in each of the individual separation compartments in the image; information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected; a calculation step of calculating the amount or concentration of the target substance based on information regarding the number of the individual separation compartments in which the target capture substance is not detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected.

2. The information processing method according to claim 1, wherein the individual separation compartment is a well or a droplet.

3. The information processing method according to claim 1, wherein the target capture substance contains particles.

4. The information processing method according to claim 3, wherein the particles contain magnetic particles.

5. The information processing method according to claim 3, wherein the particles have a particle size of 1 μm or more and 10 μm or less.

6. The information processing method according to claim 1, wherein the target capture substance contains at least one of nucleic acid, antibody, antigen, enzyme, and enzyme substrate.

7. The information processing method according to claim 1, wherein the target substance contains at least one of nucleic acid, antibody, antigen, enzyme, and enzyme substrate.

8. The information processing method according to claim 1, wherein the image includes at least one of a bright field image and a fluorescence image.

9. The information processing method according to claim 1, wherein the volume of the individual separation compartment is 0.1 fL or more and 1000 fL or less.

10. The calculation step in claim 1 includes a step of performing negative and positive determination based on the relationship between a predetermined threshold value and the luminance value of the individual separation compartment in which the target capture substance is detected, and the relationship between the predetermined threshold value and the luminance value of the individual separation compartment in which the target capture substance is not detected.

11. The calculation step in claim 1 includes a step of calculating the amount or concentration of the target substance using the average number of target substances in the individual separation compartments in which the target capture substance is detected and the average number of target substances in the individual separation compartments in which the target capture substance is not detected.

12. A program for causing the information processing method according to claim 1 to be executed.

13. A placement unit for placing an aggregate including a plurality of individual separation compartments; An imaging unit for imaging an image including a plurality of the individual separation compartments as subjects; An information processing unit, and includes: The information processing unit: An image acquisition unit that acquires an image including a plurality of individual separation compartments that may include at least one of a target substance and a target capture substance that captures the target substance as subjects; A detection unit that detects the presence or absence of the capture substance in each of the individual separation compartments in the image; Information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected; A calculation unit that calculates the amount or concentration of the target substance based on the number of the individual separation compartments in which the target capture substance is not detected and information regarding the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected; A calculation system having the above.

14. An information processing system including an information processing unit that processes an image including a plurality of individual separation compartments as subjects, The information processing unit: An image acquisition unit that acquires an image including a plurality of individual separation compartments that may include at least one of a target substance and a target capture substance that captures the target substance as subjects; A detection unit that detects the presence or absence of the capture substance in each of the individual separation compartments in the image; Information regarding the number of the individual separation compartments in which the target capture substance is detected and the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is detected; A calculation unit that calculates the amount or concentration of the target substance based on the number of the individual separation compartments in which the target capture substance is not detected and information regarding the number of the compartments in which the target substance is detected among the individual separation compartments in which the target capture substance is not detected; An information processing system having the above.

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