Droplet control device, droplet production method, and composition

Abstract

A droplet control device for controlling droplets produced from a polysaccharide and a polyether, the droplet control device comprising: a light irradiation unit for irradiating light; and a monitoring unit for monitoring the state of the droplets. An aqueous medium including the droplets further includes a photoisomerizable compound. The light irradiation unit irradiates the aqueous medium with light of a wavelength that photoisomerizes the photoisomerizable compound.

Description

DROPLET CONTROL DEVICE, METHOD FOR MANUFACTURING DROPLETS, AND COMPOSITIONS

[0001] The present invention relates to a droplet control device, a droplet production method, and a composition. The present invention also relates to an apparatus for separating a target substance and a method for separating a target substance. This application claims priority to Japanese Patent Application No. 2023-181844, filed in Japan on October 23, 2023, the contents of which are incorporated herein by reference.

[0002] Liquid-liquid phase separation droplets are known as a technique for providing a closed space in water that maintains fluidity. For example, by using droplets with a diameter of 10 nm, a volume of 0.5 yL (0.5 × 10 －２４ It is possible to construct an ultra-small reaction space of about L. Using a two-phase system of dextran and polyethylene glycol, it is possible to inexpensively and very simply prepare a large number of droplets. However, droplets are generally unstable, and within a few minutes, droplets spontaneously fuse together, resulting in macroscale phase separation.

[0003] As a technique utilizing a two-phase system of dextran and polyethylene glycol, a method for forming droplets on a femtoliter reactor array and performing a digital bioassay has been reported (Non-Patent Document 1).

[0004] Y. Minagawa et al., On-Chip Enrichment System for Digital Bioassay Based on Aqueous Two-Phase System. ACS Nano 2023, 17, 1, 212-220.

[0005] In conventional liquid-liquid phase separation droplet technology, droplets are unstable and difficult to control. Therefore, it is difficult to use droplets as a site for chemical reactions or biological phenomena. Furthermore, it is difficult to selectively recover only the desired droplets after an assay when droplets are formed in a microspace such as a femtoliter reactor array. Furthermore, it can be difficult to selectively recover a target substance from a test composition containing multiple substances.

[0006] Therefore, an object of the present invention is to provide a droplet control device and a droplet production method capable of controlling liquid-liquid phase separation droplets. Another object of the present invention is to provide a composition capable of controlling the initiation of a reaction of multiple components. Another object of the present invention is to provide an apparatus for separating a target substance from a sample and a method for separating a target substance from a sample.

[0007] The present invention includes the following aspects. [1] A droplet control device for controlling droplets made from a polysaccharide and a polyether, comprising: a light irradiation unit for irradiating light; and a monitoring unit for monitoring the state of the droplets, wherein an aqueous medium containing the droplets further contains a photoisomerizable compound, and the light irradiation unit irradiates the aqueous medium with light of a wavelength that photoisomerizes the photoisomerizable compound. [2] The droplet control device according to [1], further comprising a processor for controlling the operation of the droplet control device. [3] The droplet control device according to [1] or [2], further comprising a container for containing the aqueous medium. [4] The droplet control device according to [2] or [3], wherein the processor controls the operation of the light irradiation unit, and the operation of the light irradiation unit controlled by the processor is at least one selected from the group consisting of timing of start of light irradiation, light irradiation duration, wavelength of irradiated light, and light irradiation position. [5] The droplet control device according to any one of [2] to [4], wherein the processor controls the operation of the monitoring unit, and the operation of the monitoring unit controlled by the processor is at least one selected from the group consisting of a monitoring start timing, a monitoring time, and a monitoring position. [6] The droplet control device according to any one of [1] to [5], further comprising an output unit for outputting a monitoring result by the monitoring unit. [7] The droplet control device according to any one of [1] to [6], wherein the monitoring unit comprises a microscope unit for magnifying and observing the droplet. [8] The droplet control device according to any one of [1] to [7], wherein the droplets include a first droplet containing a first component and a second droplet containing a second component, and the first component is a component that interacts with the second component.[9] The droplet control device described in [8], further comprising a first component storage section for storing the first component, a second component storage section for storing the second component, a polysaccharide storage section for storing a polysaccharide, a polyether storage section for storing a polyether, and a photoisomerizable compound storage section for storing the photoisomerizable compound.

[10] The droplet control device according to [9], further comprising a droplet preparation unit, wherein the droplet preparation unit is controlled to mix an aqueous solution containing the first component supplied from the first component storage unit, the polysaccharide supplied from the polysaccharide storage unit, the polyether supplied from the polyether storage unit, and the photoisomerizable compound supplied from the photoisomerizable compound storage unit to prepare the first droplet, and to mix an aqueous solution containing the second component supplied from the second component storage unit, the polysaccharide supplied from the polysaccharide storage unit, the polyether supplied from the polyether storage unit, and the photoisomerizable compound supplied from the photoisomerizable compound storage unit to prepare the second droplet.

[11] The droplet control device according to

[10] , further comprising a detection reagent storage unit for supplying a detection reagent, wherein the detection reagent storage unit is controlled to supply the detection reagent to the droplet preparation unit at either or both of the timings when the first droplet is prepared and the second droplet is prepared.

[12] The droplet control device according to

[10] or

[11] , wherein the droplet production unit includes a stirring mechanism.

[13] The droplet control device according to any one of [8] to

[12] , wherein the monitoring unit is controlled to monitor an interaction between the first component and the second component in a third droplet produced by fusing the first droplet and the second droplet.

[14] The droplet control device according to any one of [8] to

[13] , wherein the monitoring unit is controlled to perform monitoring based on a time point when the light irradiation unit starts irradiating the aqueous medium with light of a wavelength at which the photoisomerizable compound photoisomerizes.

[15] The droplet control device according to any one of [8] to

[14] , wherein the first component is a cell, the second component is a virus, and the interaction is infection of the cell by the virus.

[16] The droplet control device according to [3], wherein the container comprises a plurality of wells and a flow path communicating with the plurality of wells.

[17] The droplet control device according to

[16] , wherein the volume of the well is 1 picoliter or less.

[18] The droplet control device according to

[16] or

[17] , further comprising a liquid-sending unit that sends liquid to the plurality of wells via the flow path.

[19] The droplet control device according to

[18] , wherein the liquid-sending unit comprises a polysaccharide storage section for supplying a polysaccharide, a polyether storage section for supplying a polyether, and a photoisomerizable compound storage section for supplying the photoisomerizable compound.

[20] The droplet control device according to

[19] , wherein the liquid-feeding unit is controlled to perform (i) feeding of an aqueous solution containing the polysaccharide to the plurality of wells, (ii) feeding of an aqueous solution containing the polyether to the plurality of wells, and (iii) feeding of an aqueous solution containing the polyether and the photoisomerizable compound to the plurality of wells in this order.

[21] The droplet control device according to

[20] , wherein the monitoring unit is controlled to start monitoring after (iii).

[22] The droplet control device according to

[21] , wherein the light irradiation unit is controlled to irradiate wells other than a specific well with light of a wavelength at which the photoisomerizable compound photoisomerizes, based on the monitoring result of the monitoring unit.

[23] The droplet control device according to

[22] , further including a droplet recovery unit for recovering the droplet from the specific well.

[24] The droplet control device according to any one of [1] to

[23] , wherein the photoisomerizable compound is a compound containing a peptide structure represented by the following general formula (p1) or (p2): XaXb(XaXc). ｎ XaXdXa ... (p1) XaXd (XaXc) ｎXaXbXa (p2) [In the formula, Xa represents an amino acid residue having a hydrophilic side chain; Xb represents an amino acid residue having a side chain represented by the following general formula (b1); Xc and Xd each independently represent an amino acid residue having a hydrophobic side chain or a glycine residue; and n represents an integer of 1 to 3. Multiple Xa's may be the same or different. When n is 2 or 3, multiple Xc's may be the same or different.]

[0008] [In the formula, Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent; Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. * represents a bond.]

[0009]

[25] An apparatus for separating a target substance from a sample prepared in an aqueous medium containing a photoisomerizable compound, the apparatus comprising: a container having a plurality of compartments for accommodating the sample; a light irradiation unit for irradiating the sample accommodated in the compartments with light; and a monitoring unit for monitoring the sample accommodated in the compartments, wherein the light irradiation unit irradiates the sample with light of a wavelength that photoisomerizes the photoisomerizable compound, and the sample containing the photoisomerizable compound undergoes a sol-gel transition due to the photoisomerization of the photoisomerizable compound.

[26] A method for preparing droplets using a droplet container having a plurality of wells and flow paths communicating with the plurality of wells, the method comprising the following steps (a), (b), and (c1) in this order, or the following steps (b), (a), and (c2) in this order: (a) delivering an aqueous solution containing a polysaccharide to the plurality of wells via the flow paths; (b) delivering an aqueous solution containing a polyether to the plurality of wells via the flow paths; (c1) delivering an aqueous solution containing the polyether and a photoisomerizable compound to the plurality of wells via the flow paths; (c2) delivering an aqueous solution containing the polysaccharide and a photoisomerizable compound to the plurality of wells via the flow paths.

[27] The method according to

[26] , further comprising, after step (c1) or (c2), (d) irradiating wells other than the specific wells with light of a wavelength that photoisomerizes the photoisomerizable compound.

[28] The method according to

[27] , further comprising, after step (d), (e1) delivering an aqueous solution containing a polyether to the plurality of wells via the flow path and recovering droplets from the specific wells, or (e2) delivering an aqueous solution containing a polysaccharide to the plurality of wells via the flow path and recovering droplets from the specific wells.

[29] A method for separating a target substance from a sample prepared in an aqueous medium containing a photoisomerizable compound, the method comprising: (A) distributing the sample prepared in a sol-state aqueous medium containing the photoisomerizable compound into a plurality of compartments; (B) irradiating (i) a compartment containing the target substance or (ii) a compartment not containing the target substance with light of a wavelength that photoisomerizes the photoisomerizable compound, thereby gelling the sample contained in the compartment irradiated with the light; and (C) removing the sample from the compartment not irradiated with the light.

[30] The method according to any one of

[26] to

[29] , wherein the photoisomerizable compound is a compound containing a peptide structure represented by the following general formula (p1) or (p2): XaXb(XaXc). ｎ XaXdXa ... (p1) XaXd (XaXc) ｎ XaXbXa (p2) [In the formula, Xa represents an amino acid residue having a hydrophilic side chain; Xb represents an amino acid residue having a side chain represented by the following general formula (b1); Xc and Xd each independently represent an amino acid residue having a hydrophobic side chain or a glycine residue; and n represents an integer of 1 to 3. Multiple Xa's may be the same or different. When n is 2 or 3, multiple Xc's may be the same or different.]

[0010] [In the formula, Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent; Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. * represents a bond.]

[0011]

[29] A composition contained in a light-shielding container, the composition comprising: a polysaccharide, a polyether, a photoisomerizable compound, a first droplet encapsulating a first component, and a second droplet encapsulating a second component.

[30] The composition according to

[29] , wherein the photoisomerizable compound is a compound comprising a peptide structure represented by the following general formula (p1) or (p2): XaXb(XaXc) ｎ XaXdXa ... (p1) XaXd (XaXc)ｎ XaXbXa (p2) [In the formula, Xa represents an amino acid residue having a hydrophilic side chain; Xb represents an amino acid residue having a side chain represented by the following general formula (b1); Xc and Xd each independently represent an amino acid residue having a hydrophobic side chain or a glycine residue; and n represents an integer of 1 to 3. Multiple Xa's may be the same or different. When n is 2 or 3, multiple Xc's may be the same or different.]

[0012] [In the formula, Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent; Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. * represents a bond.]

[0013]

[31] A multiple-component formulation comprising an openable, light-shielding container and the composition according to

[29] or

[30] contained in the light-shielding container.

[0014] The present invention provides a droplet control device and a droplet production method capable of controlling liquid-liquid phase separation droplets. It also provides a composition capable of controlling the initiation of a reaction of multiple components. The present invention also provides an apparatus for separating a target substance from a sample and a method for separating a target substance from a sample.

[0015] 1 is a schematic diagram showing a configuration example of a droplet control device of a first embodiment; FIG. 1 is a schematic diagram showing a configuration example of a microscopy unit, which is an example of a monitoring unit; FIG. 2 is a flowchart showing an operation example of a droplet control device of a first embodiment; FIG. 3 is a schematic diagram showing a state of a photoisomerizable compound and droplets before light irradiation in a droplet control device of a first embodiment; FIG. 4 is a schematic diagram showing a state of a photoisomerizable compound and droplets during light irradiation in a droplet control device of a first embodiment; FIG. 5 is a schematic diagram showing a state of a photoisomerizable compound and droplets after light irradiation in a droplet control device of a first embodiment; FIG. 6 is a schematic diagram showing a configuration example of a droplet control device of a second embodiment; FIG. 7 is a schematic diagram showing a configuration example of a container of a droplet control device of a second embodiment; FIG. 8 is a schematic diagram showing a modified example of a container of a droplet control device of a second embodiment; FIG. 9 is a flowchart showing an operation example of a droplet control device of a second embodiment; FIG. 10 is a schematic diagram explaining a procedure for producing droplets in a droplet control device of a second embodiment; FIG. 11 is a diagram showing a state in which an aqueous polysaccharide solution has been delivered; FIG. 12 is a schematic diagram explaining a procedure for producing droplets in a droplet control device of a second embodiment; 1 shows a state in which an aqueous polyether solution has been delivered. FIG. 1 is a schematic diagram illustrating a procedure for producing droplets in the droplet control device of the second embodiment. FIG. 2 shows a state in which an aqueous polyether solution containing a photoisomerizable compound has been delivered. FIG. 3 is a schematic diagram illustrating a procedure for collecting droplets in the droplet control device of the second embodiment. FIG. 4 shows a state in which wells other than the specific well are irradiated with light. FIG. 5 is a schematic diagram illustrating a procedure for collecting droplets in the droplet control device of the second embodiment. FIG. 6 shows a state in which wells other than the specific well are irradiated with light. FIG. 7 is a schematic diagram illustrating a procedure for collecting droplets in the droplet control device of the second embodiment. FIG. 8 shows a state in which an aqueous polyether solution has been delivered. FIG. 9 is a flowchart illustrating another example of operation of the droplet control device of the second embodiment. FIG. 10 is a flowchart illustrating an example of operation when target cells are recovered using the droplet control device of the second embodiment. FIG. 11 is a schematic diagram illustrating a procedure for producing droplets containing cells in the droplet control device of the second embodiment. FIG. 12 shows a state in which an aqueous polysaccharide solution has been delivered. FIG. 13 is a schematic diagram illustrating a procedure for producing droplets containing cells in the droplet control device of the second embodiment. This shows the state in which a polyether aqueous solution containing cells is delivered.1 is a schematic diagram illustrating a procedure for producing droplets containing cells in a droplet control device of a second embodiment. FIG. 2 shows a state in which an aqueous polyether solution containing a photoisomerizable compound has been delivered. FIG. 3 is a schematic diagram illustrating a procedure for recovering droplets containing target cells in a droplet control device of a second embodiment. FIG. 4 shows a state in which light is irradiated onto wells other than the wells containing the target cells. FIG. 5 is a schematic diagram illustrating a procedure for recovering droplets containing target cells in a droplet control device of a second embodiment. FIG. 6 shows a state after light has been irradiated onto wells other than the wells containing the target cells. FIG. 7 is a schematic diagram illustrating a procedure for recovering droplets containing target cells in a droplet control device of a second embodiment. FIG. 8 shows a state in which an aqueous polyether solution has been delivered. FIG. 9 is a flowchart illustrating an example of operation when a second component that reacts with a first component is recovered using a droplet control device of a second embodiment. FIG. 10 is a schematic diagram illustrating a procedure for producing droplets containing a first component and a second component in a droplet control device of a second embodiment. FIG. 11 shows a state in which an aqueous polysaccharide solution containing the first component has been delivered. FIG. 12 is a schematic diagram illustrating a procedure for producing droplets containing a first component and a second component in a droplet control device of a second embodiment. 1 shows a state in which an aqueous polyether solution containing a second component has been delivered. FIG. 1 is a schematic diagram illustrating a procedure for preparing droplets containing the first component and the second component in a droplet control device of a second embodiment. FIG. 2 shows a state in which an aqueous polyether solution containing a photoisomerizable compound has been delivered. FIG. 3 is a schematic diagram illustrating a procedure for recovering droplets containing the first component and the second component in a droplet control device of a second embodiment. FIG. 4 shows a state in which wells other than the wells containing the first component and the second component are irradiated with light. FIG. 5 is a schematic diagram illustrating a procedure for recovering droplets containing the first component and the second component in a droplet control device of a second embodiment. FIG. 6 shows a state after wells other than the wells containing the first component and the second component are irradiated with light. FIG. 7 is a schematic diagram illustrating a procedure for recovering droplets containing the first component and the second component in a droplet control device of a second embodiment. FIG. 8 shows a state in which an aqueous polyether solution has been delivered. FIG. 9 is a schematic diagram illustrating a state in which droplet A and droplet B coexist in Example 1. Droplet A contains E. coli and fluorescein-di-β-D-galactopyranoside (FDG), and droplet B contains M13 phage.21(a)-21(c) are images showing the results of detecting green fluorescence from fluorescein when a solution containing droplets A and B is irradiated with and without light irradiation in Example 1. FIG. 21(a) is a schematic diagram showing the procedure of Example 2. FIG. 21(b) is an image showing the state of the well before the aqueous PEG solution is delivered (before (4) in FIG. 19) and before the aqueous PEG solution is delivered (after (4) in FIG. 19) in Example 2. FIG. 21(c) is an image showing the state of the well when the aqueous PEG solution is delivered without light irradiation (after (4) in FIG. 21) and when the aqueous PEG solution is delivered with light irradiation (after (6) in FIG. 21) in Example 3. FIG. 21(c) is a schematic diagram showing an example of the configuration of an apparatus for separating a target substance according to one embodiment. FIG. 21(c) is a flow chart showing an example of the operation when a target substance is recovered using an apparatus for separating a target substance according to one embodiment.

[0016] Hereinafter, embodiments of the present invention will be described in detail, with reference to the drawings where necessary. In the drawings, identical or corresponding parts are designated by identical or corresponding reference numerals, and duplicate explanations will be omitted. The dimensional ratios in each drawing may be exaggerated for the purpose of explanation and do not necessarily correspond to the actual dimensional ratios.

[0017] A numerical range expressed using "to" means a range that includes the numerical values ​​before and after "to" as the lower and upper limits.

[0018] Unless otherwise specified, "a," "an," and "the" are inclusive of singular and plural and are understood to mean "one or more."

[0019] The term "comprise" means that it may contain components other than the target component. The term "consist of" means that it does not contain components other than the target component. The term "consist essentially of" means that it does not contain components other than the target component in a manner that would exert a special function (such as a manner that would completely lose the effects of the invention). In this specification, when "comprise" is used, it includes both "consist of" and "consist essentially of" embodiments.

[0020] [Droplet Control Device] A first aspect of the present disclosure is a droplet control device for controlling droplets made from a polysaccharide and a polyether. In one embodiment, the droplet control device includes a light irradiation unit for irradiating light and a monitoring unit for monitoring the state of the droplets. The aqueous medium containing the droplets further includes a photoisomerizable compound. The light irradiation unit is used to irradiate the aqueous medium with light of a wavelength that photoisomerizes the photoisomerizable compound.

[0021] <Droplets> The droplets controlled by the droplet control device of the embodiment are droplets made of a polysaccharide and a polyether (hereinafter referred to as "droplet D"). When aqueous solutions containing a polysaccharide and a polyether are mixed and stirred, minute droplets containing a high concentration of polysaccharide are typically formed by liquid-liquid phase separation. Alternatively, as described below, droplets containing a high concentration of polysaccharide are formed in the wells by feeding an aqueous polysaccharide solution followed by an aqueous polyether solution into a flow path device having wells formed in the flow paths. Alternatively, droplets containing a high concentration of polyether are formed in the wells by feeding an aqueous polyether solution followed by an aqueous polysaccharide solution into a flow path device having wells formed in the flow paths.

[0022] When droplet D is coexisted with a photoisomerizable compound, the state of droplet D can be controlled by changing the photoisomerization state of the photoisomerizable compound.

[0023] (Polysaccharides) The polysaccharide is not particularly limited. Examples of polysaccharides include, but are not limited to, dextran, dextrin, amylose, amylopectin, glycogen, cellulose, agarose, carrageenan, heparin, alginic acid, hyaluronic acid, pectin, glucomannan, tamarind seed gum, guar gum, locust bean gum, gum arabic, karaya gum, xanthan gum, and gellan gum. The polysaccharide preferably contains glucose as a constituent sugar, and is preferably dextran or dextrin, with dextran being more preferred.

[0024] The molecular weight of the polysaccharide is not particularly limited, but examples thereof include 10,000 to 10,000,000, 50,000 to 5,000,000, 100,000 to 5,000,000, 300,000 to 1,000,000, and 400,000 to 800,000.

[0025] The polysaccharide may be labeled with a fluorescent dye or the like. The fluorescent dye is not particularly limited, and known fluorescent dyes can be used. To avoid droplet fusion, it is preferable to use a fluorescent dye having an excitation wavelength and a fluorescence wavelength different from the wavelength at which cis-conversion of compound (P) occurs. For example, when compound (P) having a side chain represented by formula (b1-1-1) described below is used, a fluorescent dye having an excitation wavelength and a fluorescence wavelength of 490 nm or longer can be used. Examples of such fluorescent dyes include, but are not limited to, rhodamine, 5-FAM, 5-FITC, ATTO520, AlexaFlour555, Cy3Cy3, Cy5, Alexa568, and Alexa647.

[0026] (Polyether) The polyether is not particularly limited. The polyether is a compound having a plurality of ether bonds in the molecule. The polyether may be an aliphatic polyether or an aromatic polyether, but an aliphatic polyether is preferred. Examples of the aliphatic polyether include polyalkylene glycol. Examples of the polyalkylene glycol include polyethylene glycol, polypropylene glycol, polytetramethylene glycol, polyoxyethylene polyoxypropylene glycol, etc. The polyether is preferably polyethylene glycol.

[0027] The molecular weight of the polyether is not particularly limited, but examples thereof include 300 to 10,000,000, 500 to 5,000,000, 1,000 to 3,000,000, 5,000 to 1,000,000, and 10,000 to 500,000.

[0028] (Photoisomerizable compound) A "photoisomerizable compound" refers to a compound that undergoes isomerization upon irradiation with light. A photoisomerizable compound reversibly changes from a trans isomer to a cis isomer or from a cis isomer to a trans isomer upon irradiation with light of a specific wavelength.

[0029] The photoisomerizable compound is preferably a compound that undergoes a sol-gel transition in an aqueous medium containing the photoisomerizable compound upon photoisomerization. In one embodiment, the photoisomerizable compound forms a gel in the trans isomer and a sol in the cis isomer. In one embodiment, the photoisomerizable compound forms a sol in the trans isomer and a gel in the cis isomer.

[0030] The photoisomerizable compound is preferably a compound having affinity for either polyether or polysaccharide. When the photoisomerizable compound has a higher affinity for polyether than for polysaccharide, the photoisomerizable compound is localized in the polyether phase in the dispersion of droplet D. When the photoisomerizable compound has a higher affinity for polysaccharide than for polyether, the photoisomerizable compound is localized in the polysaccharide phase in the dispersion of droplet D. It is preferable to use a photoisomerizable compound that has a high affinity for compounds localized outside the droplet. Examples of photoisomerizable compounds that have a high affinity for polyether include the compound (P) described below.

[0031] (Aqueous Medium) The "aqueous medium" is an aqueous liquid used to dissolve the polysaccharide, polyether, and photoisomerizable compound when preparing Droplet D. Examples of aqueous media include water, various buffer solutions (phosphate-buffered saline, acetate buffer, phosphate buffer, citrate buffer, citrate-phosphate buffer, borate buffer, tartrate buffer, Tris buffer, HEPES buffer, etc.), and various liquid media (Dulbecco's modified Eagle's medium (D-MEM) medium, MEMα medium, etc.). The aqueous medium preferably has a pH of 4 to 10, more preferably a pH of 5 to 9, more preferably a pH of 6 to 9, and particularly preferably a pH of 7 to 9.

[0032] Droplet D is formed by mixing the polysaccharide and the polyether in an aqueous medium, and the formed droplet D is retained in the aqueous medium.

[0033] (First Component, Second Component) When preparing droplet D by mixing an aqueous polysaccharide solution and an aqueous polyether solution, a desired component can be contained in these aqueous solutions to form droplet D in which the desired component is encapsulated within droplet D. The component to be encapsulated in droplet D is not particularly limited, and examples thereof include organic synthetic compounds such as low molecular weight compounds, fluorescent dyes, radical generators, and synthetic polymer compounds; biomolecules such as amino acids, nucleotides, sugars, vitamins, peptides, proteins, and nucleic acids; inorganic compounds such as inorganic nanoparticles, inorganic anions, inorganic cations, inorganic salts, and aggregates thereof; cells such as animal cells, plant cells, fungi, algae cells, and protozoan cells; microstructures such as phages, viruses, and molecular aggregates; and cell aggregates such as organoids and spheroids.

[0034] For example, a first droplet D1 containing a first component and a second droplet D2 containing a second component can be separately prepared and then mixed and fused to form a third droplet D3 containing the first and second components. If the second component interacts with the first component, the interaction between the first and second components can be monitored over time, with the formation of the third droplet D3 being the starting point of the interaction. Examples of combinations of a first component and a second component include a cell and an infectious virus for that cell, a compound that reacts with the cell, an antibody and an antigen, an active substance and an inhibitor, etc.

[0035] In the following description, a droplet containing the first component will be referred to as a first droplet D1, a droplet containing the second component will be referred to as a second droplet D2, and a droplet formed by fusion of the first droplet D1 and the second droplet D2 will be referred to as a third droplet D3. These may also be collectively referred to as droplet D.

[0036] <First embodiment> (Configuration example of droplet control device) Figure 1 is a schematic diagram showing a configuration example of a droplet control device of the first embodiment. The droplet control device 1 includes a container 10, a monitoring unit 20, a light irradiation unit 30, an output unit 50, an input unit 51, a communication unit 52, a memory 53, a waste liquid storage unit 68, and a droplet preparation unit 70, as well as a processor 40 for controlling the operation of each of these units. The droplet control device 1 also includes storage units for various components, including a polysaccharide storage unit 61, a polyether storage unit 62, a photoisomerizable compound storage unit 63, a first component storage unit 64, a second component storage unit 65, a detection reagent storage unit 66, and an aqueous medium storage unit 67.

[0037] In the droplet control device 1, components for producing the first droplet D1 are supplied to the droplet production unit 70 from the polysaccharide storage unit 61, the polyether storage unit 62, the photoisomerizable compound storage unit 63, and the first component storage unit 64. The droplet production unit 70 mixes and stirs these components to produce the first droplet D1. The produced first droplet D1 is transferred to the container 10 together with the aqueous medium. Furthermore, components for producing the second droplet D2 are supplied to the droplet production unit 70 from the polysaccharide storage unit 61, the polyether storage unit 62, the photoisomerizable compound storage unit 63, and the second component storage unit 65. The droplet production unit 70 mixes and stirs these components to produce the second droplet D2. The produced second droplet D2 is transferred to the container 10 together with the aqueous medium.

[0038] An aqueous medium containing first droplet D1 and second droplet D2 contained in container 10 is irradiated with light from light irradiation unit 30. As a result, the photoisomerizable compound contained in the aqueous medium is photoisomerized, and the first droplet D1 and the second droplet D2 fuse to form a third droplet D3. In the third droplet D3, the first component and the second component coexist, and as the third droplet D3 is formed, an interaction between these components begins. A monitoring unit 20 monitors the states of the first droplet D1, the second droplet D2, and the third droplet D3 in container 10. The monitoring unit 20 observes the interaction between the first component and the second component in the third droplet D3 over time.

[0039] <Container> The container 10 is for containing an aqueous medium containing droplet D. The container 10 is not particularly limited, but preferably has an internal space surrounded by a bottom surface, sidewall surfaces, and a ceiling surface. The container 10 may have a ceiling surface formed by an openable lid, or may be a closed type in which a ceiling plate is fixed to the sidewall. Alternatively, the container 10 may be an open type without a ceiling surface.

[0040] The material of the container 10 is not particularly limited, but if the container 10 has a ceiling surface, the ceiling surface is made of a material that can transmit light irradiated from the light irradiation unit 30. Furthermore, if the monitoring unit 20 includes a microscopic unit described below, the entire container 10 may be made of a transparent, low-autofluorescence material. Examples of transparent, low-autofluorescence materials include glass, polyethylene terephthalate (PET), polycarbonate, cycloolefin polymer, polydimethylsiloxane, polystyrene, and polyacrylate (acrylic resin).

[0041] The size of the container 10 is not particularly limited, but the depth of the droplet storage space is preferably 1 mm or less. When the depth of the droplet storage space is 1 cm or less, overlapping of the droplets D in the vertical direction is suppressed, making it easier to observe the droplets D using the monitoring unit 20. Examples of the depth of the droplet storage space include 10 nm to 0.5 mm, 0.1 mm to 50 μm, and 10 μm or less. The lower limit of the depth of the droplet storage space is not particularly limited, but examples include 10 nm to 50 nm, 100 nm to 500 nm, and 1 μm or more. Examples of the depth range of the droplet storage space include 10 nm to 1 mm, 10 nm to 0.5 mm, 10 nm to 0.1 mm, 10 nm to 50 μm, and 10 nm to 10 μm. The "droplet storage space" is the internal space of the container 10 defined by the bottom surface, sidewall surfaces, and ceiling surface. If the container 10 does not have a ceiling surface, the "droplet-receiving space" is the space defined by the bottom surface and the sidewall surface.

[0042] The container 10 is equipped with a temperature controller 10t for controlling the temperature of the droplet accommodation space. The temperature controller 10t controls the temperature of the droplet accommodation space to a temperature suitable for the interaction between the first component and the second component. The temperature controller 10t may be a heater, a cooler, a thermostat, or the like.

[0043] The container 10 may be detachable. For example, the droplet control device 1 may include a container holder, and a container prepared by the user may be placed in the container holder as the container 10.

[0044] <<Monitoring Unit>> The monitoring unit 20 is for monitoring the state of the droplets D contained in the container 10. The monitoring unit 20 is not particularly limited as long as it can observe the state of the droplets D.

[0045] The monitoring unit 20 may include, for example, a unit equipped with a microscope unit for magnifying and observing the droplet D. FIG. 2 shows an example of the microscope unit. The microscope unit 21 is disposed, for example, so as to face the lower surface of the container 10. The microscope unit 21 may be any microscope, such as a bright-field microscope, a phase-contrast microscope, a differential interference microscope, or a fluorescence microscope. The microscope unit 21 may also be a digital microscope.

[0046] The microscope unit 21 includes an objective lens 22. The objective lens 22 is disposed so as to face the bottom surface of the container 10. The objective lens 22 creates a magnified image of the droplet D in the container 10 from light incident on the objective lens 22 from the container 10. Any known objective lens can be used as the objective lens 22.

[0047] The image sensor 28 acquires a magnified image of the inside of the container 10 formed by the microscope unit 21 as image data and outputs the acquired image data. The image sensor 28 may be provided inside the microscope unit 21, or may be arranged outside the microscope unit 21 so as to receive the output of the magnified image from the microscope unit 21. For example, the microscope unit 21 may be a digital microscope incorporating the image sensor 28. Any image sensor such as a CMOS camera or a CCD camera can be used as the image sensor 28.

[0048] The light source 27 is arranged to emit light parallel to the optical axis of the objective lens 22 toward the control mirror 26, for example. Any light source, such as a laser device, can be used as the light source 27. The light emitted from the light source 27 is used to observe the droplets D in the container 10 via the objective lens 22. The wavelength of the light emitted by the light source 27 can be selected appropriately depending on the state of the droplets D to be observed. For example, when the fluorescence intensity changes as the interaction in the third droplet D3 progresses, the light source 27 may emit excitation light to detect the fluorescence.

[0049] The wavelength of the light emitted by the light source 27 may be the same as or different from the wavelength that photoisomerizes the photoisomerizable compound. From the viewpoint of strictly controlling the photoisomerization of the photoisomerizable compound, the wavelength of the light emitted by the light source 27 is preferably different from the wavelength that photoisomerizes the photoisomerizable compound.

[0050] The microscope unit 21 may include a control mirror 26, a pupil projection lens 25, an imaging lens 24, a dichroic mirror 23, and the like to direct light from the light source 27 into the objective lens 22. The control mirror 26 includes one or more mirrors and a driver for driving the mirror. The control mirror 26 can adjust the optical path of the reflected light by controlling and driving the position and tilt of the mirror at high speed. Examples of the control mirror 26 include, but are not limited to, electromagnetically driven mirrors such as a galvanometer mirror, a digital mirror device (DMD), and a MEMS mirror. The pupil projection lens 25 is disposed on the optical path of the light reflected by the control mirror 26. The pupil projection lens 25 converts the light pattern radiating from the control mirror 26 into parallel light. Any lens, such as a general convex lens, can be used as the pupil projection lens 25 as long as it satisfies the above functions. The imaging lens 24 is disposed downstream of the pupil projection lens 25 on the optical path of the light reflected by the control mirror 26. The imaging lens 24 focuses the light pattern converted into parallel rays by the pupil projection lens 25 at the back focal position of the objective lens 22. Any lens can be used as the imaging lens 24, such as a general convex lens, as long as it satisfies the above functions. The dichroic mirror 23 is provided below the objective lens 22 and is arranged downstream of the imaging lens 24 on the optical path of the light reflected by the control mirror 26. The dichroic mirror 23 reflects only light of a specific wavelength (for example, wavelengths in the near-infrared region) and transmits light of other wavelengths. The dichroic mirror 23 reflects light of a wavelength output by the light source 27. The dichroic mirror 23 reflects the light pattern formed by the light source 27 and the control mirror 26 toward the objective lens 22.

[0051] The monitoring unit 20 is not limited to one including the microscope unit 21, and may be one that observes the droplets in the container 10 by other methods. The monitoring unit 20 may also include, for example, an analyzer unit. The analyzer unit may be the same as the analyzer of a flow cytometer. The analyzer unit may include a channel through which the droplets pass one by one, a light source that irradiates the droplets passing through the channel, and a photodetector that detects scattered light from the droplets.

[0052] <<Light Irradiation Unit>> The light irradiation unit 30 is for irradiating the aqueous medium containing the droplets D in the container 10 with light. The light irradiation unit 30 has a mechanism for irradiating the aqueous medium with light of a wavelength that photoisomerizes the photoisomerizable compound. The wavelength at which the photoisomerizable compound photoisomerizes varies depending on the structure of the photoisomerizable compound. Therefore, the wavelength of light irradiated by the light irradiation unit 30 is controlled depending on the type of photoisomerizable compound.

[0053] When the photoisomerizable compound is in a gel state in the trans isomer and in a sol state in the cis isomer, it is preferable that the light irradiated by the light irradiation unit 30 has a wavelength that photoisomerizes the photoisomerizable compound from the trans isomer to the cis isomer. For example, when the compound (P) described below has a side chain represented by the formula (b1-1-1) described below as the side chain (b1), examples of the wavelength of the irradiation light that causes the cis conversion of the compound (P) include 200 to 400 nm, 250 to 390 nm, 300 to 380 nm, 320 to 370 nm, and 330 to 360 nm, and for example, irradiation light of 350 nm can be used. When compound (P) has a side chain represented by formula (b1-1-7) described below as the side chain (b1), examples of the wavelength of the irradiation light that causes the cis-formation of compound (P) include 400 to 700 nm, 450 to 650 nm, 500 to 600 nm, 510 to 580 nm, 520 to 570 nm, and 530 to 560 nm. For example, irradiation light of 540 nm can be used.

[0054] The light irradiation unit 30 may be equipped with a light source that generates light. Examples of light sources include lasers, continuous spectrum lamps (e.g., mercury vapor lamps, halogen lamps, and tungsten amplifiers), and light-emitting diodes (LEDs). Examples of lasers include solid-state lasers such as YAG lasers, ruby ​​lasers, glass lasers, YVO4 lasers, LD lasers, and fiber lasers; and semiconductor lasers. The light irradiation unit 30 may be equipped with multiple light sources that generate light of different wavelengths, and may be able to switch between these light sources depending on the type of photoisomerizable compound. Alternatively, the light irradiation unit 30 may be equipped with a tunable wavelength laser.

[0055] The light irradiation unit may include an optical member for adjusting the wavelength and direction of light emitted from the light source, such as a control mirror, a dichroic mirror, a bandpass filter, a mirror, a collimating lens, or a spectroscope.

[0056] <Droplet Preparation Unit> The droplet preparation unit 70 is used to prepare droplets D. The droplet preparation unit 70 mixes and stirs the polysaccharide aqueous solution, the polyether aqueous solution, the photoisomerizable compound aqueous solution, and the first component supplied from the polysaccharide storage section 61, the polyether storage section 62, the photoisomerizable compound storage section 63, and the first component storage section 64, respectively, to prepare first droplets D1. The droplet preparation unit 70 also mixes and stirs the polysaccharide aqueous solution, the polyether aqueous solution, the photoisomerizable compound aqueous solution, and the second component supplied from the polysaccharide storage section 61, the polyether storage section 62, the photoisomerizable compound storage section 63, and the second component storage section 65, respectively, to prepare second droplets D2.

[0057] The droplet preparation unit 70 includes a mixing tank for mixing and stirring the components, and a stirring mechanism 71 for stirring the liquid in the mixing tank. The stirring mechanism 71 is not particularly limited, but examples thereof include a mechanism for rotating a stirrer having a rod-like, plate-like, or propeller-like shape, a mechanism for rotating a stirring blade fixed in the mixing tank, and a mechanism for rotating the mixing tank itself.

[0058] The droplet preparation unit 70 may include a temperature controller for controlling the temperature in the mixing tank. Examples of the temperature controller include a heater, a cooler, and a thermostat.

[0059] A waste liquid storage section 68 is connected to the droplet preparation unit 70. The waste liquid storage section 68 stores waste liquid used to wash the inside of the mixing tank after droplets D prepared in the mixing tank are transferred to the container 10, for example. The inside of the mixing tank is washed with an aqueous medium supplied from an aqueous medium storage section 67, for example. The connection and disconnection between the droplet preparation unit 70 and the waste liquid storage section 68 is switched by a valve B8.

[0060] <<Individual Component Storage Units>> The polysaccharide storage unit 61 is for storing an aqueous polysaccharide solution. The concentration of the aqueous polysaccharide solution stored in the polysaccharide storage unit 61 is preferably 0.3% by mass or more. In the mixed solution containing various components used in the preparation of droplet D, the polysaccharide concentration may be 0.1 to 40% by mass relative to the total mass of the mixed solution. The concentration of the aqueous polysaccharide solution stored in the polysaccharide storage unit 61 may be any concentration that allows the polysaccharide concentration to be adjusted to within the above concentration range when the mixed solution is prepared. The aqueous polysaccharide solution stored in the polysaccharide storage unit 61 may be a concentrated solution, and in this case, it may be diluted by supplying an aqueous medium from the aqueous medium storage unit 67. The polysaccharide storage unit 61 is connected to the droplet preparation unit 70 via a conduit 90. The connection and disconnection between the polysaccharide storage unit 61 and the droplet preparation unit 70 is switched by a valve B1.

[0061] The polyether container 62 is for containing an aqueous polyether solution. The concentration of the aqueous polyether solution contained in the polyether container 62 is preferably 0.3% by mass or more. In the mixed solution containing various components used in the preparation of droplet D, the polyether concentration may be 0.1 to 40% by mass relative to the total mass of the mixed solution. The concentration of the aqueous polyether solution contained in the polyether container 62 may be any concentration that allows the polyether concentration to be adjusted to within the above concentration range when the mixed solution is prepared. The aqueous polyether solution contained in the polyether container 62 may be a concentrated solution, and in this case, it may be diluted by supplying an aqueous medium from the aqueous medium container 67. The polyether container 62 is connected to the droplet preparation unit 70 by a conduit 90. The connection and disconnection between the polyether container 62 and the droplet preparation unit 70 are switched by valves B1 and B2.

[0062] The photoisomerizable compound storage section 63 is for storing an aqueous photoisomerizable compound solution. The concentration of the aqueous photoisomerizable compound solution stored in the photoisomerizable compound storage section 63 is preferably 0.3% by mass or more. In the mixed solution containing various components used in the preparation of droplet D, the concentration of the photoisomerizable compound may be 0.1 to 10% by mass relative to the total mass of the mixed solution. The concentration of the aqueous photoisomerizable compound solution stored in the photoisomerizable compound storage section 63 may be any concentration that allows the concentration of the photoisomerizable compound to be adjusted within the above-mentioned concentration range when the mixed solution is prepared. The aqueous photoisomerizable compound solution stored in the photoisomerizable compound storage section 63 may be a concentrated solution, and in this case, it may be diluted by supplying an aqueous medium from the aqueous medium storage section 67. The photoisomerizable compound storage section 63 is connected to the droplet preparation unit 70 via a conduit 90. The connection and disconnection between the photoisomerizable compound storage section 63 and the droplet preparation unit 70 is switched by valves B1, B2, and B3. The photoisomerizable compound stored in the photoisomerizable compound storage section 63 is preferably an isomer that forms a gel state. For example, in the case of a photoisomerizable compound whose trans isomer is in a gel state and whose cis isomer is in a sol state, the trans isomer is preferably stored in the photoisomerizable compound storage section 63. In the case of compound (P) described below, it is preferable that the trans isomer of compound (P) be stored in the photoisomerizable compound storage section 63.

[0063] The first component storage unit 64 is for storing a liquid (solution or dispersion) containing a first component. The liquid can be selected depending on the type of the first component. Examples of the medium for the liquid include those listed above as aqueous media. The concentration of the first component in the liquid stored in the first component storage unit 64 is not particularly limited and can be set appropriately depending on the type of the first component. The concentration of the first component in the liquid stored in the first component storage unit 64 may be a concentration that allows the concentration of the first component to be adjusted to a desired concentration range when made into a mixed solution. The liquid containing the first component stored in the first component storage unit 64 may be a concentrated liquid, and in this case, it may be diluted by supplying an aqueous medium from the aqueous medium storage unit 67. The first component storage unit 64 is connected to the droplet preparation unit 70 by a conduit 90. The first component storage unit 64 and the droplet preparation unit 70 are connected and disconnected by valves B1, B2, B3, and B4.

[0064] The second component storage unit 65 is for storing a liquid (solution or dispersion) containing the second component. The liquid can be selected depending on the type of the second component. Examples of the liquid medium include those similar to those listed above as the aqueous medium. The concentration of the second component in the liquid stored in the second component storage unit 65 is not particularly limited and can be set appropriately depending on the type of the second component. The concentration of the second component in the liquid stored in the second component storage unit 65 may be a concentration that, when mixed into a mixed solution, allows the concentration of the second component to be adjusted to a desired concentration range. The liquid containing the second component stored in the second component storage unit 65 may be a concentrated liquid, in which case it may be diluted by supplying an aqueous medium from the aqueous medium storage unit 67. The second component storage unit 65 is connected to the droplet preparation unit 70 via a conduit 90. The second component storage unit 65 and the droplet preparation unit 70 are connected and disconnected by valves B1, B2, B3, B4, and B5.

[0065] The detection reagent storage unit 66 is for storing an aqueous detection reagent solution. The detection reagent is a reagent capable of detecting an interaction between a first component and a second component. The detection reagent may be supplied to the droplet preparation unit 70 during the preparation of the first droplet D1, or may be supplied to the droplet preparation unit 70 during the preparation of the second droplet D2. Alternatively, the detection reagent may be supplied to the droplet preparation unit 70 both during the preparation of the first droplet D1 and the preparation of the second droplet D2. The aqueous detection reagent solution stored in the detection reagent storage unit 66 may be a concentrated solution, in which case it may be diluted by supplying an aqueous medium from the aqueous medium storage unit 67. The detection reagent storage unit 66 is connected to the droplet preparation unit 70 by a conduit 90. The connection and disconnection between the detection reagent storage unit 66 and the droplet preparation unit 70 are switched by valves B1, B2, B3, B4, B5, and B6.

[0066] The aqueous medium storage section 67 is for storing an aqueous medium. The aqueous medium is used for diluting the mixed solution when producing droplets D and for cleaning the mixing tank of the droplet production unit 70 after producing droplets D. The aqueous medium storage section 67 is connected to the droplet production unit 70 by a conduit 90. The connection and disconnection between the aqueous medium storage section 67 and the droplet production unit 70 is switched by valves B1, B2, B3, B4, B5, B6, and B7.

[0067] <<Output Unit, Input Unit, Communication Unit, Memory>> The droplet control device 1 includes an output unit 50 , an input unit 51 , and a communication unit 52 .

[0068] The output unit 50 outputs various information such as the monitoring results of the monitoring unit 20. The output unit 50 includes, for example, a display, a speaker, etc. The display displays information. For example, the display displays various information output from the processor 40 as images, or displays a GUI (Graphical User Interface) for receiving various input operations from the user. For example, the display is an LCD (Liquid Crystal Display), an organic EL (Electro Luminescence) display, etc. The speaker outputs the information output by the processor 40 as sound.

[0069] The input unit 51 accepts various input operations from the user, converts the accepted input operations into electrical signals, and outputs the electrical signals to the processor 40. The input unit 51 includes physical operation components such as a mouse, keyboard, trackball, switches, buttons, joystick, and touch panel. The input unit 51 may also be a user interface that accepts audio input from a microphone or the like. When the input unit 51 is a touch panel, the input unit 51 may also have a display function such as a display.

[0070] The input unit 51 is not limited to those having physical operation parts such as a mouse, a keyboard, etc. For example, an example of the input unit 51 also includes an electrical signal processing circuit that receives an electrical signal corresponding to an input operation from an external input device provided separately from the device and outputs this electrical signal to a control circuit.

[0071] The communication unit 52 includes, for example, a network interface card (NIC) and an antenna for wireless communication. The communication unit 52 communicates with external devices via a communication network NW. The communication network NW may refer to any information communication network that uses electrical communication technology. For example, the communication network NW includes a local area network (LAN), a wide area network (WAN), the Internet, a telephone communication network, an optical fiber communication network, a cable communication network, a satellite communication network, and the like.

[0072] The external device may be, for example, a computer connected to a LAN (also called a workstation), or a cloud server connected to a WAN or the Internet.

[0073] The memory 53 is realized by, for example, a semiconductor memory element such as a RAM (Random Access Memory), a flash memory, a hard disk, or an optical disk. These non-transitory storage media may be realized by other storage devices connected via a communication network NW, such as a NAS (Network Attached Storage) or an external storage server device. The memory 53 may also include other non-transitory storage media such as a ROM (Read Only Memory) or a register.

[0074] <<Processor>> The processor 40 controls the operation of the droplet control device 1. The operation of each of the above units of the droplet control device 1 is controlled by the processor 40. The processor 40 realizes these controls by, for example, executing a program stored in the memory 53.

[0075] The processor 40 may be, for example, a central processing unit (CPU), a graphics processing unit (GPU), an application specific integrated circuit (ASIC), a programmable logic device (e.g., a simple programmable logic device (SPLD) or a complex programmable logic device (CPLD)), or a field programmable gate array (FGA). The program may be a circuit (circuitry) such as a FPGA (Field Programmable Gate Array). Instead of storing the program in memory 53, the program may be directly embedded in the circuit of processor 40. In this case, processor 40 realizes its functions by reading and executing the program embedded in the circuit. The program may be stored in memory 53 in advance, or may be stored in a non-transitory storage medium such as a DVD or CD-ROM, and installed into memory 53 from the non-transitory storage medium by inserting the non-transitory storage medium into a drive device (not shown) of droplet control device 1. Processor 40 is not limited to being configured as a single circuit, but may be configured as a single hardware processor by combining multiple independent circuits to realize each function. Furthermore, multiple components may be integrated into a single hardware processor to realize each function.

[0076] The processor 40 has a container control unit 41, a light irradiation unit control unit 42, a droplet production unit control unit 43, a monitoring unit control unit 44, an output unit control unit 45, an input unit control unit 46, and a communication unit control unit 47, and has the function of controlling the operation of each of the above units.

[0077] The container control unit 41 has a function of controlling the container 10. The container control unit 41 may have a function of controlling the temperature controller 10t to control the temperature of the droplet accommodation space.

[0078] The light irradiation unit control unit 42 has a function of controlling the light irradiation unit 30. The light irradiation unit control unit 42 may have a function of controlling, for example, at least one operation of the light irradiation unit 30 selected from the group consisting of the timing of starting light irradiation, the light irradiation time, the wavelength of the irradiated light, and the light irradiation position.

[0079] The droplet preparation unit control unit 43 has a function of controlling the droplet preparation unit 70. It may also have a function of controlling the supply of each component from the polysaccharide storage section 61, the polyether storage section 62, the photoisomerizable compound storage section 63, the first component storage section 64, the second component storage section 65, the detection reagent storage section 66, and the aqueous medium storage section 67 to the droplet preparation unit 70. For example, the droplet preparation unit control unit 43 may control the supply of these components to the droplet preparation unit 70 by controlling valves B1 to B7. The droplet preparation unit control unit 43 may also have a function of controlling the stirring mechanism 71 of the droplet preparation unit 70. For example, the droplet preparation unit control unit 43 may have a function of controlling the operation timing, stirring time, stirring intensity, etc. of the stirring mechanism 71. The droplet preparation unit control unit 43 may also have a function of controlling the transfer of droplets D from the droplet preparation unit 70 to the container 10. For example, the droplet production unit control unit 43 may control the transfer of droplets D from the droplet production unit 70 to the container 10 by controlling valve B9. The droplet production unit control unit 43 may have a function of controlling the cleaning of the mixing tank of the droplet production unit 70. For example, the droplet production unit control unit 43 may have a function of controlling the timing of supply of aqueous medium from the aqueous medium storage unit 67 to the droplet production unit 70 and the discharge of waste liquid to the waste liquid storage unit 68. For example, the droplet production unit control unit 43 may control the cleaning of the droplet production unit 70 by controlling valves B1 to B8. The droplet production unit control unit 43 may have a function of controlling the temperature in the mixing tank of the droplet production unit 70.

[0080] The monitoring unit control unit 44 has a function of controlling the monitoring unit 20. The monitoring unit control unit 44 may have a function of controlling at least one operation of the monitoring unit 20 selected from the group consisting of the timing of starting monitoring, the monitoring time, and the monitoring position, for example.

[0081] The output unit control section 45 has a function of outputting various information such as the monitoring results of the monitoring unit 20 via the output unit 50 .

[0082] The input unit control section 46 has a function of processing information input from the input unit 51 .

[0083] The communication unit control unit 47 has a function of controlling communication via the communication unit 52 .

[0084] (Example of Operation of Droplet Control Device) FIG. 3 is a flowchart showing an example of operation of the droplet control device 1.

[0085] In step S1, the polysaccharide storage section 61, the polyether storage section 62, the photoisomerizable compound storage section 63, and the first component storage section 64 supply liquids containing a polysaccharide aqueous solution, a polyether aqueous solution, a photoisomerizable compound aqueous solution, and a first component to the droplet preparation unit 70, respectively. At this time, the amount of each component required for preparation of the first droplet D1 is supplied. If necessary, a detection reagent may be supplied to the droplet preparation unit 70 from the detection reagent storage section 66. Alternatively, if necessary, an aqueous medium may be supplied to the droplet preparation unit 70 from the aqueous medium storage section 67. The amount of each component supplied may be preset or may be set by the user via the input unit 51. The supply of each component to the droplet preparation unit 70 may be performed by a pump (not shown) or the like.

[0086] In step S2, the mixed solution of each component supplied to the mixing tank of the droplet preparation unit 70 is mixed and stirred to prepare the first droplet D1. The stirring time may be set in advance or may be set by the user via the input unit 51. Alternatively, the stirring time may be manually stopped by the user via the input unit 51.

[0087] In step S3, the first droplets D1 prepared in step S2 are contained in the container 10. The transfer of the first droplets D1 from the droplet preparation unit 70 to the container 10 is performed by controlling the valve B9. The entire amount of the aqueous medium containing the first droplets D1 may be transferred to the container 10, or a portion may be transferred to the container 10 and the remainder may be stored in the waste liquid storage section 68.

[0088] The aqueous medium containing the first droplets D1 may be transferred to the container 10 or the waste liquid storage section 68 by a pump or the like (not shown).

[0089] In step S4, the polysaccharide storage section 61, the polyether storage section 62, the photoisomerizable compound storage section 63, and the second component storage section 65 supply liquids containing a polysaccharide aqueous solution, a polyether aqueous solution, a photoisomerizable compound aqueous solution, and a second component, respectively, to the droplet preparation unit 70. At this time, the amount of each component required for preparation of the second droplet D2 is supplied. If necessary, a detection reagent may be supplied to the droplet preparation unit 70 from the detection reagent storage section 66. Alternatively, if necessary, an aqueous medium may be supplied to the droplet preparation unit 70 from the aqueous medium storage section 67. The amount of each component supplied may be preset or may be set by the user via the input unit 51. The supply of each component to the droplet preparation unit 70 may be performed by a pump (not shown) or the like. After step S3 and before step S4, an aqueous medium may be supplied from the aqueous medium storage section 67 to wash the mixing chamber of the droplet preparation unit 70.

[0090] In step S5, the mixed solution of each component supplied to the mixing tank of the droplet preparation unit 70 is mixed and stirred to prepare a second droplet D2. The stirring time may be set in advance or may be set by the user via the input unit 51. Alternatively, the stirring time may be manually stopped by the user via the input unit 51.

[0091] In step S6, the second droplet D2 produced in step S5 is contained in the container 10. The transfer of the second droplet D2 from the droplet production unit 70 to the container 10 is performed by controlling the valve B9. The entire amount of the aqueous medium containing the second droplet D2 may be transferred to the container 10, or a portion may be transferred to the container 10 and the remainder may be stored in the waste liquid storage section 68. By step S6, the aqueous medium containing the first droplet D1 and the second droplet D2 is held in the container 10.

[0092] In step S7, the monitoring unit 20 starts monitoring the droplets D in the container 10. The monitoring status may be displayed on a display or the like, which is the output unit 50. Furthermore, monitoring images or videos may be recorded as needed in the memory 53 or an external storage device.

[0093] In step S8, the light irradiation unit 30 begins irradiating the aqueous medium containing droplets D in the container 10 with light. The wavelength of the light irradiated at this time is a wavelength that photoisomerizes the photoisomerizable compound. The wavelength and irradiation time of light may be preset depending on the type of photoisomerizable compound, or may be set by the user via the input unit 51. The irradiation time may be a time sufficient to photoisomerize the photoisomerizable compound. Examples of the irradiation time include approximately 0.1 seconds to 15 minutes. The irradiation time may be 0.1 seconds to 15 minutes, 0.5 seconds to 15 minutes, 1 second to 15 minutes, 5 seconds to 15 minutes, or 10 seconds to 15 minutes.

[0094] In step S9, the light irradiation unit 30 ends the light irradiation.

[0095] In step S10, the monitoring unit 20 ends monitoring.

[0096] In step S11, the monitoring result is output via the output unit 50. This completes the series of operation flows.

[0097] In the above operational example, the order of steps S7 and S8 may be reversed. The order of steps S9 and S10 may be reversed. The start time of monitoring in step 7 may be set to a predetermined time before the start of light irradiation, simultaneously with the start of light irradiation, or after a predetermined time has elapsed since the start of light irradiation, based on the time when the light irradiation unit starts irradiating the aqueous medium with light.

[0098] 4A to 4C are schematic diagrams illustrating the behavior of the photoisomerizable compound and droplet D in step S8.

[0099] 4A shows the state before light irradiation. Before light irradiation, the photoisomerizable compound PI forms a fibrous structure. When the photoisomerizable compound PI is compound (P), compound (P) is in a trans form before light irradiation. Because the photoisomerizable compound PI forms a fibrous structure, the aqueous medium W is in a gel state. As a result, the first droplet D1 containing the first component C1 and the second droplet D2 containing the second component C2 are maintained in the aqueous medium W without fusing with each other.

[0100] FIG. 4B shows a state in which light is irradiated from the monitoring unit 20. The light from the monitoring unit 20 isomerizes the photoisomerizable compound PI to form a photoisomerizable compound PI', which is an isomer of the photoisomerizable compound PI. When the photoisomerizable compound PI is compound (P), the photoisomerizable compound PI' is a cis-isomer of compound (P). The fibrous structure of the photoisomerizable compound PI' collapses and the photoisomerizable compound PI' disperses in the aqueous medium W. This causes the aqueous medium W to become a sol. This facilitates fusion of the first droplet D1 and the second droplet D2 within the aqueous medium W. Therefore, upon initiation of light irradiation, fusion of the first droplet D1 and the second droplet D2 begins. This also initiates interaction between the first component C1 and the second component C2.

[0101] 4C shows the state after light irradiation. After light irradiation, a third droplet D3 is formed by the fusion of the first droplet D1 and the second droplet D2. The third droplet D3 encapsulates the first component C1 and the second component C2. The interaction between the first component C1 and the second component C2 progresses inside the third droplet D3.

[0102] The droplet control device of the first embodiment, having the above-described configuration, can control the start time of the interaction between the first component and the second component by light irradiation, thereby enabling the interaction of multiple components to be monitored over time from the start of the interaction.

[0103] (Modification) The droplet control device may include two or more droplet preparation units. In this case, the first droplet D1 and the second droplet D2 may be prepared in separate droplet preparation units. The droplet control device may include a container for holding an aqueous medium containing the first droplet D1 and the second droplet D2 installed within the device. In this case, the containers for each component, the droplet preparation units, and the containers may not be provided in the droplet control device. The droplet control device may include a holder for holding a container installed by the user.

[0104] Second Embodiment (Configuration Example of Droplet Control Device) Figure 5 is a schematic diagram showing a configuration example of a droplet control device of the second embodiment. The droplet control device 100 includes a container 110, a monitoring unit 120, a light irradiation unit 130, an output unit 150, an input unit 151, a communication unit 152, a memory 153, a waste liquid storage unit 168, and a droplet recovery unit 170, as well as a processor 140 for controlling the operation of each of these units. The droplet control device 100 also includes storage units for various components, including a polysaccharide storage unit 161, a polyether storage unit 162, a photoisomerizable compound storage unit 163, a first component storage unit 164, a second component storage unit 165, a detection reagent storage unit 166, and an aqueous medium storage unit 167.

[0105] The droplet control device 100 does not have a droplet preparation unit, and is configured so that each component is sent from each component storage section 161 to 167 to a container 110. In other words, the container 110 also serves as a droplet preparation unit. A waste liquid storage section 168 and a droplet recovery unit 170 are connected to the container 110 via a conduit 190. Other than the above configuration, the droplet control device 100 has the same configuration as the droplet control device 1.

[0106] <Container> FIG. 6A is a schematic diagram showing an example of a container 110. The container 110 includes a plurality of wells 111 and a channel 115 communicating with the plurality of wells 111. The container 110 is composed of a ceiling substrate 110a, a bottom substrate 110b, and a sidewall 110c. The sidewall 110c is disposed between the ceiling substrate 110a and the bottom substrate 110b and defines the height of the internal space of the container 110. The sidewall 110c is fixed to the edges of the ceiling substrate 110a and the bottom substrate 110b. As a result, the interior of the container 110 is a closed space with only the supply port 116 and the discharge port 117 open. The height of the internal space of the container 110 defined by the sidewall 110c can be, for example, 1 nm or more, 10 nm or more, 100 nm or more, 1 μm or more, or 10 μm or more. The upper limit of the height is, for example, 5 mm or less, 1 mm or less, 500 μm or less, 100 μm or less, or 50 μm or less. The range of the height is, for example, 1 nm to 5 mm or less, 10 nm to 1 mm or less, 100 nm to 500 μm or less, 1 μm to 100 μm or less, or 10 μm to 50 μm or less.

[0107] A plurality of wells 111 separated by partition walls 112 are provided on the inner bottom surface of the bottom substrate 110b. A flow path 115 is formed above each well 111, and the plurality of wells 111 communicate with each other via the flow path 115. Each well 111 is a recess with one open end, and is a space defined by the partition wall 112 and the inner surface of the bottom substrate 110b. The shape of the well 111 is not particularly limited, and examples thereof include a cylindrical shape and a polygonal prism shape (such as a square prism or a hexagonal prism).

[0108] The volume of the well 111 is not particularly limited as long as it is large enough to form droplets D. Examples of the volume of the well 111 include 10 nanoliters or less, 1 nanoliter or less, 100 picoliters or less, 10 picoliters or less, 1 picoliters or less, 100 femtoliters or less, and 50 femtoliters or less. The lower limit of the volume of the well 111 is not particularly limited, and examples include 1 attoliter or more, 10 attoliters or more, 100 attoliters or more, or 1 femtoliter or more. Examples of the volume range of the well 111 include 1 attoliter or more and 10 nanoliters or less, 10 attoliters or more and 100 picoliters or less, 100 attoliters or more and 10 picoliters or less, or 1 femtoliter or more and 1 picoliter or less.

[0109] The side wall 110c is provided with a supply port 116 and a discharge port 117, each of which is connected to a conduit 190. The liquid supplied from the supply port 116 to the inside of the container 110 via the conduit 190 passes through the flow path 115 and is discharged from the discharge port 117. During this process, the liquid is filled into the multiple wells 111 via the flow path 115. As will be described later, droplets D are formed in the wells 111 by changing the type of liquid delivered to the inside of the container 110 in a predetermined order.

[0110] Fig. 6B shows a modified example of the container 110. The container 110' has the same configuration as the container 110 shown in Fig. 6A, except that a plurality of wells 111' are provided on the inner surface of a top substrate 110a' rather than on a bottom substrate 110b'.

[0111] The material of the container 110 is not particularly limited, but is preferably a material that transmits the light irradiated by the light irradiation unit 130. Furthermore, when the monitoring unit 120 includes a microscope unit, the entire container 110 may be made of a transparent material with low autofluorescence. Examples of transparent materials with low autofluorescence include the same materials as those described above.

[0112] The container 110 can be manufactured by, for example, photolithography microfabrication technology, 3D printer technology, or the like.

[0113] The container 110 is equipped with a temperature controller 110t for controlling the temperature of the internal space. The temperature controller 110t controls the temperature of the internal space depending on the component to be encapsulated in the droplet D. The temperature controller 110t may be a heater, a cooler, a thermostat, or the like.

[0114] The container 110 may be detachable. For example, the droplet control device 100 may include a container holder, and a container prepared by the user may be placed in the container holder as the container 110.

[0115] <Liquid Delivery Unit> The droplet control device 100 includes a liquid delivery unit 160 that delivers liquid to the internal space of the container 110. The liquid delivery unit 160 delivers liquid to the multiple wells 111 via the flow path 115. The liquid delivery unit 160 is composed of a polysaccharide storage section 161, a polyether storage section 162, a photoisomerizable compound storage section 163, a first component storage section 164, a second component storage section 165, a detection reagent storage section 166, an aqueous medium storage section 167, a waste liquid storage section 168, a droplet recovery unit 170, and a pump P.

[0116] The configurations of the polysaccharide storage section 161, the polyether storage section 162, the photoisomerizable compound storage section 163, the first component storage section 164, the second component storage section 165, the detection reagent storage section 166, and the detection reagent storage section 166 are the same as those in the first embodiment. Instead of the photoisomerizable compound aqueous solution, the photoisomerizable compound storage section 163 may store an aqueous solution containing a photoisomerizable compound and a polyether (photoisomerizable compound / polyether aqueous solution) or an aqueous solution containing a photoisomerizable compound and a polysaccharide (photoisomerizable compound / polysaccharide aqueous solution). When the photoisomerizable compound has a high affinity for polyether, the photoisomerizable compound storage section 163 preferably stores a photoisomerizable compound / polyether aqueous solution. When the photoisomerizable compound has a high affinity for polysaccharide, the photoisomerizable compound storage section 163 preferably stores a photoisomerizable compound / polysaccharide aqueous solution.

[0117] The aqueous polysaccharide solution contained in the polysaccharide storage unit 161 is supplied into the interior of the container 110 via the conduit 190 by switching the valve B1. The aqueous polysaccharide solution is pumped into the interior of the container 110 by the driving force of the pump P. The aqueous polysaccharide solution that flows into the interior of the container 110 from the supply port 116 passes through the flow path 115, is discharged from the discharge port 117, and is stored in the waste liquid storage unit 168. The connection state between the container 110 and the waste liquid storage unit 168 is switched by the valves B8 and B9. When the aqueous polysaccharide solution contained in the polysaccharide storage unit 161 is a concentrated solution, the aqueous polysaccharide solution may be diluted in the conduit 190 by supplying an aqueous medium from the aqueous medium storage unit 167.

[0118] The other components are similarly delivered to the interior of the container 110 .

[0119] The liquid-feeding unit 160 may be controlled to perform, in this order, (i) the delivery of the aqueous polysaccharide solution to the plurality of wells 111, (ii) the delivery of the polyether solution to the plurality of wells 111, and (iii) the delivery of the aqueous photoisomerizable compound / polyether solution to the plurality of wells 111. Alternatively, the liquid-feeding unit 160 may be controlled to perform, in this order, (i) the delivery of the aqueous polyether solution to the plurality of wells 111, (ii) the delivery of the polysaccharide solution to the plurality of wells 111, and (iii) the delivery of the aqueous photoisomerizable compound / polysaccharide solution to the plurality of wells 111.

[0120] The droplet recovery unit 170 is for recovering droplets D held in any of the wells 111. The container 110 and the droplet recovery unit 170 are connected via a conduit 190. The connection state between the container 110 and the droplet recovery unit 170 is switched by valves B8 and B9.

[0121] <Monitoring Unit> The monitoring unit 120 is for monitoring the state of the droplets in the container 110. The monitoring unit 120 may include the above-described microscope unit. In one embodiment, the monitoring unit 120 is controlled to start monitoring after (iii) above.

[0122] <Light Irradiation Unit> The light irradiation unit 130 is for irradiating light onto the aqueous medium containing the droplets D in the container 110. As a light source for the light irradiation unit 130, the same one as the light irradiation unit 30 in the first embodiment can be used.

[0123] The light irradiation unit 130 includes a mechanism for irradiating light onto wells 111, excluding specific wells 111, among the multiple wells 111 in the container 110. The light irradiation unit 130 may include, for example, a member that blocks light irradiated from a light source onto the specific wells 111. An example of the light blocking member is a photomask. Alternatively, the light irradiation unit 130 may include a laser device that can control the light irradiation position.

[0124] The output unit 150, input unit 151, and communication unit 152 can be the same as those in the first embodiment.

[0125] <<Processor>> The processor 140 controls the operation of the droplet control device 100. The operation of each of the above units of the droplet control device 100 is controlled by the processor 40. The processor 40 realizes these controls by, for example, executing a program stored in the memory 53. The hardware configuration of the processor 140 can be the same as that in the first embodiment.

[0126] The processor 140 has a container control unit 141, a light irradiation unit control unit 142, a liquid delivery unit control unit 143, a monitoring unit control unit 144, an output unit control unit 145, an input unit control unit 146, and a communication unit control unit 147, and has the function of controlling the operation of each of the above units.

[0127] The container control unit 141 has a function of controlling the container 110. The container control unit 141 may have a function of controlling the temperature of the internal space of the container 110 by controlling the temperature controller 110t.

[0128] The light irradiation unit control unit 142 has a function of controlling the light irradiation unit 130. The light irradiation unit control unit 142 may have a function of controlling at least one operation selected from the group consisting of the timing of starting light irradiation, the light irradiation time, the wavelength of the irradiation light, and the light irradiation position in the light irradiation unit 130, for example. The light irradiation unit control unit 142 may have a function of controlling a photomask. For example, the light irradiation unit control unit 142 may have a function of selecting wells 111 to block the irradiation light based on the monitoring information of the monitoring unit 120, and placing a photomask at the position of the specific wells 111. Alternatively, the light irradiation unit control unit 142 may have a function of controlling a laser device included in the light irradiation unit 130 to irradiate wells 111 other than the specific wells 111 with laser light.

[0129] The liquid-sending unit control unit 143 has a function of controlling the liquid-sending unit 160. The liquid-sending unit control unit 143 may have a function of controlling the sending of each component from the polysaccharide storage unit 161, the polyether storage unit 162, the photoisomerizable compound storage unit 163, the first component storage unit 164, the second component storage unit 165, the detection reagent storage unit 166, and the aqueous medium storage unit 167 to the container 110. For example, the liquid-sending unit control unit 143 may control the sending of these components to the container 110 by controlling the valves B1 to B7 and the pump P. The liquid-sending unit control unit 143 may have a function of controlling the order in which the liquid is sent from each component storage unit. The liquid-sending unit control unit 143 may have a function of controlling the speed at which the liquid is sent from each component storage unit.

[0130] The liquid sending unit control section 143 may have a function of controlling the valves B8 and B9 to switch the destination of the liquid discharged from the container 110 to either the droplet recovery unit 170 or the waste liquid storage section 168.

[0131] The monitoring unit control unit 144 has a function of controlling the monitoring unit 20. The monitoring unit control unit 44 may have a function of controlling at least one operation of the monitoring unit 20 selected from the group consisting of the timing of starting monitoring, the monitoring time, and the monitoring position, for example.

[0132] The output unit control section 145, the input unit control section 146, and the communication unit control section 47 are the same as those in the first embodiment.

[0133] (Operational Example of Droplet Control Device) Figure 7 is a flowchart showing an operational example of the droplet control device 100. Figure 7 shows an operational example when using a photoisomerizable compound that has a higher affinity for polyether than for polysaccharide. Figures 8A to 8C and 9A to 9C are schematic diagrams explaining the state inside the container 110 at each step.

[0134] In step S101, the aqueous polysaccharide solution is delivered from the polysaccharide storage section 161 to the container 110. The delivery of the aqueous polysaccharide solution is performed by the liquid delivery unit control section 143 controlling the valve B1 and the pump P. The delivery of the aqueous polysaccharide solution continues until all of the wells 111 in the container 110 are filled with the aqueous polysaccharide solution. At this time, the valves B8 and B9 are controlled so that the container 110 is connected to the waste liquid storage section 168. The delivery time may be set in advance or may be set by the user via the input unit 151. Alternatively, a sensor or the like installed in the container 110 may detect the filling state of the container 110 and control the stopping of the delivery. The delivery rate of the aqueous polysaccharide solution is not particularly limited as long as it is a rate at which the aqueous polysaccharide solution fills the wells 111. The delivery rate of the aqueous polysaccharide solution may be adjusted appropriately depending on the sizes of the container 110 and the wells 111. When the size of the well 111 is on the order of femtoliters, the speed of delivery of the aqueous polysaccharide solution is, for example, 1 to 1000 μL / min, or may be 10 to 800 μL / min, or may be 50 to 500 μL / min. Fig. 8A shows a state in which the flow path 115 and well 111 in the container 110 are filled with the aqueous polysaccharide solution PS as a result of delivery of the aqueous polysaccharide solution PS. The arrows in the figure indicate the delivery direction.

[0135] In step S102, the aqueous polyether solution is delivered from the polyether storage section 162 to the container 110. The delivery of the aqueous polyether solution is performed by the delivery unit control section 143 controlling the valves B1 and B2 and the pump P. The delivery of the aqueous polyether solution continues until the flow path 115 in the container 110 is filled with the aqueous polyether solution. The delivery time may be set in advance or may be set by the user via the input unit 151. Alternatively, a sensor or the like installed in the container 110 may detect the formation state of droplets D in the multiple wells 111 and control the cessation of delivery. The delivery rate of the aqueous polyether solution is not particularly limited as long as it is a rate at which the aqueous polysaccharide solution in the flow path 115 is replaced with the aqueous polyether solution. The delivery rate of the aqueous polyether solution may be adjusted appropriately depending on the sizes of the container 110 and the wells 111. When the size of the well 111 is on the order of femtoliters, the rate of delivery of the aqueous polyether solution is, for example, 1 to 1,000 μL / min, or may be 10 to 800 μL / min, or may be 50 to 500 μL / min. Fig. 8B shows a state in which the flow path 115 in the container 110 is filled with the aqueous polyether solution PE as a result of delivery of the aqueous polyether solution PE. At this time, the aqueous polysaccharide solution PS remains in the well 111, and droplets D are formed in the well 111.

[0136] In step S103, the photoisomerizable compound / polyether aqueous solution is sent from the photoisomerizable compound storage section 163 to the container 110. The sending of the photoisomerizable compound / polyether aqueous solution from the photoisomerizable compound storage section 163 is performed by the liquid sending unit control section 143 controlling the valves B1 to B3 and the pump P. Alternatively, the photoisomerizable compound aqueous solution may be sent from the photoisomerizable compound storage section 163, and the polyether aqueous solution may be sent from the polyether storage section 162, and they may be mixed in the conduit 190 and sent to the container 110. The sending of the polyether aqueous solution from the polyether storage section 162 and the sending of the photoisomerizable compound from the photoisomerizable compound storage section 163 is performed by the liquid sending unit control section 143 controlling the valves B1 to B3 and the pump P. The photoisomerizable compound / polyether aqueous solution is fed until the flow path 115 in the container 110 is filled with the photoisomerizable compound / polyether aqueous solution. The feeding time may be set in advance or may be set by the user via the input unit 151. Alternatively, a sensor or the like installed in the container 110 may detect the viscosity state of the liquid in the flow path 115 and control the stopping of the feeding. The feeding rate of the photoisomerizable compound / polyether aqueous solution is not particularly limited as long as it is the rate at which the polyether aqueous solution in the flow path 115 is replaced with the photoisomerizable compound / polyether aqueous solution. The feeding rate of the photoisomerizable compound / polyether aqueous solution may be appropriately adjusted depending on the size of the container 110 and the well 111. When the size of the well 111 is on the order of femtoliters, the feeding rate of the polyether aqueous solution may be, for example, 1 to 1,000 μL / min, 10 to 800 μL / min, or 50 to 500 μL / min.

[0137] 8C shows a state in which the flow path 115 in the container 110 is filled with an aqueous polyether solution PE containing the photoisomerizable compound PI as a result of the delivery of the photoisomerizable compound / aqueous polyether solution. At this time, droplet D remains in the well 111. The photoisomerizable compound PI forms a fibrous structure. When the photoisomerizable compound PI is compound (P), the trans isomer of compound (P) is delivered.

[0138] In step S104, the monitoring unit 120 starts monitoring the droplets D in the container 110. The monitoring status may be displayed on a display or the like, which is the output unit 150. Furthermore, monitoring images or videos may be recorded as needed in the memory 153 or an external storage device.

[0139] In step S105, the light irradiation unit 130 irradiates light onto wells 111 other than the specific well 111S. The wavelength of the light irradiated at this time is a wavelength that photoisomerizes the photoisomerizable compound. The light irradiation time may be a time sufficient for the photoisomerization of the photoisomerizable compound. Examples of the light irradiation time include approximately 0.1 seconds to 15 minutes. The light irradiation time may be 0.1 seconds to 15 minutes, 0.5 seconds to 15 minutes, 1 second to 15 minutes, 5 seconds to 15 minutes, or 10 seconds to 15 minutes.

[0140] The specific well 111S to which light irradiation is not performed is selected based on monitoring information from the monitoring unit 120. For example, the monitoring unit 120 detects a substance contained in a droplet D and selects a well 111 holding a droplet D containing a desired substance as the specific well. For example, a light-shielding member such as a mask is controlled to be placed in the specific well 111S, and light irradiation is performed, thereby irradiating wells other than the specific well 111S with light. Alternatively, light may be irradiated to wells 111 other than the specific well 111S using a laser device or the like capable of controlling the light irradiation position.

[0141] 9A shows a state in which light is being irradiated onto wells 111 other than the specific well 111S. A mask M is placed between the light irradiation unit 130 and the ceiling substrate 110a of the container 110 at the position of the specific well 111S, blocking light irradiation onto the specific well 111S.

[0142] FIG. 9B shows the change in the photoisomerizable compound PI upon light irradiation. The photoisomerizable compound PI present at the irradiated position undergoes photoisomerization, becoming a photoisomerizable compound PI', which is an isomer of the photoisomerizable compound PI. The photoisomerizable compound PI' loses its fibrous structure and disperses within the aqueous polyether solution PE. On the other hand, the photoisomerizable compound PI present inside and around the specific well 111S is not photoisomerized and remains as the photoisomerizable compound PI. When the photoisomerizable compound PI is compound (P), the photoisomerizable compound PI' is the cis form of compound (P).

[0143] In step S106, the light irradiation unit 130 ends the light irradiation.

[0144] In step S107, the aqueous polyether solution is delivered from the polyether storage section 162 to the container 110. The delivery of the aqueous polyether solution is performed by the delivery unit control section 143 controlling the valves B1 to B3 and the pump P. At this time, the valves B8 and B9 are controlled so that the container 110 is connected to the droplet recovery unit 170. The delivery of the aqueous polyether solution is continued until the recovery of droplets D from the specific well 111S is completed. The delivery rate of the aqueous polyether solution is not particularly limited, as long as the droplets D in the wells 111 irradiated with light in step S105 remain and the droplets D in the wells 111 not irradiated with light flow out. The delivery rate of the aqueous polyether solution may be adjusted appropriately depending on the sizes of the container 110 and the wells 111. When the size of the well 111 is on the order of femtoliters, the delivery rate of the aqueous polyether solution may be, for example, 1 to 1,000 μL / min, 10 to 800 μL / min, or 50 to 500 μL / min. FIG. 9C shows a state in which droplets D in the specific well 111S flow out of the specific well 111S due to delivery of the aqueous polyether solution PE. The photoisomerizable compound PI remains in a fibrous form inside and around the specific well 111S. Delivery of the aqueous polyether solution PE causes droplets D in the specific well 111S to flow out of the well together with the fibrous photoisomerizable compound PI. The droplets D that flow out of the specific well 111S are discharged from the outlet 117 and collected in the droplet collection unit 170 via the conduit 190.

[0145] In step 108, the monitoring unit 120 ends the monitoring.

[0146] In step S109, the monitoring result is output via the output unit 150. This completes the series of operation flows.

[0147] In the above operation example, the order of steps S106 and S107 may be reversed, and step S106 may be performed after step S108.

[0148] <Another Operation Example (1)> Fig. 10 is a flowchart showing another operation example of the droplet control device 100. Fig. 10 shows an operation example when using a photoisomerizable compound that has a higher affinity for polysaccharides than for polyethers.

[0149] In step S111, the aqueous polyether solution is sent from the polyether storage section 162 to the container 110.

[0150] In step S112 , the aqueous polysaccharide solution is transferred from the polysaccharide storage section 161 to the container 110 .

[0151] In step S113, the photoisomerizable compound / polysaccharide aqueous solution is delivered from the photoisomerizable compound storage unit 163 to the container 110. Alternatively, the photoisomerizable compound aqueous solution may be delivered from the photoisomerizable compound storage unit 163, and the polyether aqueous solution may be delivered from the polysaccharide storage unit 161, and they may be mixed in the conduit 190 and delivered to the container 110.

[0152] In step S114, the monitoring unit 120 starts monitoring the droplets D in the container 110.

[0153] In step S115, the light irradiation unit 130 irradiates light onto wells 111 other than the specific well 111S. The wavelength of the light irradiated at this time is a wavelength that photoisomerizes the photoisomerizable compound. The selection of the specific well 111S and the avoidance of light irradiation on the specific well 111S are controlled in the same manner as in step S105 above. The photoisomerizable compound PI present at the irradiated position is photoisomerized to become a photoisomerizable compound PI', which is an isomer of the photoisomerizable compound PI. On the other hand, the photoisomerizable compound PI present inside and around the specific well 111S is not photoisomerized and remains as the photoisomerizable compound PI.

[0154] In step S116, the light irradiation unit 130 ends the light irradiation.

[0155] In step S117, the aqueous polysaccharide solution is delivered from the polysaccharide storage section 161 to the container 110. At this time, the valves B8 and B9 are controlled so that the container 110 is connected to the droplet recovery unit 170. The delivery of the aqueous polysaccharide solution continues until the recovery of the droplet D from the specific well 111S is completed.

[0156] In step 118, the monitoring unit 120 ends the monitoring.

[0157] In step S119, the monitoring results are output via the output unit 150. This completes the series of operation flows. This operation flow is the same as the operation flow in Figure 7, except that the polysaccharide phase and the polyether phase are reversed.

[0158] <Another Operation Example (2)> Figure 11 is a flowchart showing another operation example of the droplet control device 100. Figure 11 shows an operation example when target cells are recovered using a photoisomerizable compound that has a higher affinity for polyether than for polysaccharide. Figures 12A to 12C and 13A to 13C are schematic diagrams explaining the state inside the container 110 at each step.

[0159] In step S121, the aqueous polysaccharide solution PS is delivered from the polysaccharide storage section 161 to the container 110. Fig. 12A shows the state in which the aqueous polysaccharide solution PS has been delivered, filling the flow path 115 and well 111 in the container 110 with the aqueous polysaccharide solution PS. The arrows in the figure indicate the delivery direction.

[0160] In step S122, the cell-containing aqueous polyether solution is delivered from the first component storage section 164 to the container 110. Alternatively, the polyether solution may be delivered from the polyether storage section 162, the cell dispersion liquid may be delivered from the first component storage section 164, and the two may be mixed in the conduit 190 and supplied into the container 110. FIG. 12B shows a state in which the channel 115 in the container 110 is filled with the aqueous polyether solution PE due to the delivery of the cell-containing aqueous polyether solution PE. At this time, the aqueous polysaccharide solution PS remains in the well 111, and droplets D are formed in the well 111. The cells include target cells Ct and non-target cells C. Because cells have a higher affinity for polysaccharides than polyethers, the cells are taken up into droplets D upon contact with the droplets D.

[0161] In step S123, the photoisomerizable compound / polyether aqueous solution is delivered from the photoisomerizable compound storage section 163 to the container 110. Alternatively, the photoisomerizable compound aqueous solution may be delivered from the photoisomerizable compound storage section 163, and the polyether aqueous solution may be delivered from the polyether storage section 162, and they may be mixed in the conduit 190 and delivered to the container 110. In step S123, a detection reagent for detecting the target cells Ct may be delivered from the detection reagent storage section 166. Examples of the detection reagent include antibodies that bind to cell membrane proteins of the target cells Ct. The antibodies may be bound to a labeling substance such as a fluorescent dye. The detection reagent delivered to the container 110 binds to the target cells Ct contained in the droplets D, making it easy to identify the target cells Ct. FIG. 12C shows a state in which the flow path 115 in the container 110 is filled with the aqueous polyether solution PE containing the photoisomerizable compound PI by feeding the photoisomerizable compound / aqueous polyether solution.

[0162] In step S124, the monitoring unit 120 starts monitoring the droplet D in the container 110. The monitoring unit 120 observes the cells taken up in the droplet D and transmits information such as the shape of the cells to the processor 140. If the detection reagent is delivered in step S123, the monitoring unit 120 may detect a labeling substance or the like bound to the detection reagent. This may obtain information on the binding state of the detection reagent to the cells and transmit the information to the processor 140.

[0163] In step S125, the processor 140 identifies the target cells Ct from among the cells taken into the droplet D based on the monitoring information transmitted from the monitoring unit 120. The processor 140 also identifies a specific well 111S that holds a droplet D containing the target cells Ct.

[0164] In step S126, the light irradiation unit 130 irradiates light onto wells 111 other than the specific well 111S containing the target cells Ct. The wavelength of the light irradiated at this time is a wavelength that photoisomerizes the photoisomerizable compound.

[0165] 13A shows a state in which light is irradiated to wells 111 other than the specific well 111S. The specific well 111S holds droplets D containing target cells Ct. The other wells 111 hold droplets containing non-target cells C or no cells.

[0166] 13B shows the change in the photoisomerizable compound PI due to light irradiation. The photoisomerizable compound PI present at the irradiated position is photoisomerized to become a photoisomerizable compound PI', which is an isomer of the photoisomerizable compound PI. The photoisomerizable compound PI present inside and around the specific well 111S is not photoisomerized and remains as the photoisomerizable compound PI.

[0167] In step S127, the light irradiation unit 130 ends the light irradiation.

[0168] In step S128, the aqueous polyether solution is delivered from the polyether storage section 162 to the container 110. At this time, valves B8 and B9 are controlled so that the container 110 is connected to the droplet recovery unit 170. Figure 13C shows a state in which droplets D containing target cells Ct in the specific well 111S are discharged from the specific well 111S due to the delivery of the aqueous polyether solution PE. The droplets D that have discharged from the specific well 111S are discharged from the outlet 117 and recovered in the droplet recovery unit 170 via the conduit 190.

[0169] In step S129, the monitoring unit 120 ends the monitoring.

[0170] In step S130, the monitoring results are output via the output unit 150. This completes the series of operation flows.

[0171] Another Operation Example (3) Figure 14 is a flowchart showing another operation example of the droplet control device 100. Figure 14 shows an operation example in which a component (second component) that reacts with an arbitrary component (first component) is recovered using a photoisomerizable compound that has a higher affinity for polyether than for polysaccharide. The first component and the second component are preferably substances that have a higher affinity for polysaccharide than for polyether. Figures 15A to 15C and 16A to 16C are schematic diagrams illustrating the state inside the container 110 at each step.

[0172] In step S131, an aqueous polysaccharide solution containing a first component is delivered from the first component storage section 164 to the container 110. Alternatively, the aqueous polysaccharide solution may be delivered from the polysaccharide storage section 161, the first component aqueous solution may be delivered from the first component storage section 164, and the two solutions may be mixed in the conduit 190 and supplied to the container 110. Fig. 15A shows a state in which the flow path 115 and well 111 in the container 110 are filled with the aqueous polysaccharide solution PS containing the first component C1 due to the delivery of the aqueous polysaccharide solution PS containing the first component C1. The arrows in the figure indicate the delivery direction.

[0173] In step S132, the aqueous polyether solution containing the second component is delivered from the second component storage section 165 to the container 110. Alternatively, the aqueous polyether solution may be delivered from the polyether storage section 162, the second component aqueous solution may be delivered from the second component storage section 165, and the two may be mixed in the conduit 190 and supplied to the container 110. FIG. 15B shows a state in which the flow path 115 in the container 110 is filled with the aqueous polyether solution PE containing the second component C2 by delivering the aqueous polyether solution PE containing the second component C2. At this time, the aqueous polysaccharide solution PS remains in the well 111, and a droplet D is formed in the well 111. At this time, the first component C1 is encapsulated in the droplet D. The second component C2, which has a binding ability to the first component C1, is incorporated into the droplet D by binding to the first component C1.

[0174] In step S133, the photoisomerizable compound / polyether aqueous solution is delivered from the photoisomerizable compound storage section 163 to the container 110. Alternatively, the photoisomerizable compound aqueous solution may be delivered from the photoisomerizable compound storage section 163, and the polyether aqueous solution may be delivered from the polyether storage section 162, and they may be mixed in the conduit 190 and delivered to the container 110. In step S123, a detection reagent for detecting the reaction between the first component C1 and the second component C2 may be delivered from the detection reagent storage section 166. Examples of the detection reagent include substances that generate fluorescence, color, or the like upon reaction between the first component and the second component. The detection reagent delivered to the container 110 generates fluorescence or color upon reaction between the first component C1 and the second component C2 contained in the droplet D, making it easier to detect the reaction between the first component C1 and the second component C2. FIG. 15C shows a state in which the flow path 115 in the container 110 is filled with the aqueous polyether solution PE containing the photoisomerizable compound PI by feeding the photoisomerizable compound / aqueous polyether solution.

[0175] In step S134, the monitoring unit 120 starts monitoring the droplet D in the container 110. The monitoring unit 120 monitors the reaction between the first component C1 and the second component C2 taken into the droplet D and transmits information such as the reaction progress state to the processor 140. If the detection reagent is delivered in step S133, the monitoring unit 120 may detect fluorescence or color development by the detection reagent. This may acquire information on the reaction state of the first component C1 and the second component C2 and transmit it to the processor 140.

[0176] In step S135, the processor 140 determines the droplet D into which the second component C2 that reacts with the first component C1 has been taken, based on the monitoring information transmitted from the monitoring unit 120. The processor 140 also identifies the specific well 111S that holds the droplet D.

[0177] In step S136, the light irradiation unit 130 irradiates light onto the wells 111 other than the specific well 111S. The wavelength of the light irradiated at this time is a wavelength that photoisomerizes the photoisomerizable compound.

[0178] 16A shows a state in which light is irradiated to wells 111 other than the specific well 111S. The specific well 111S holds a droplet D containing a second component C2 that reacts with a first component C1.

[0179] 16B shows the change in the photoisomerizable compound PI due to light irradiation. The photoisomerizable compound PI present at the irradiated position is photoisomerized to become a photoisomerizable compound PI', which is an isomer of the photoisomerizable compound PI. The photoisomerizable compound PI present inside and around the specific well 111S is not photoisomerized and remains as the photoisomerizable compound PI.

[0180] In step S137, the light irradiation unit 130 ends the light irradiation.

[0181] In step S138, the aqueous polyether solution is delivered from the polyether storage section 162 to the container 110. At this time, valves B8 and B9 are controlled so that the container 110 is connected to the droplet recovery unit 170. Figure 16C shows a state in which droplets D containing the second component C2 in the specific well 111S flow out of the specific well 111S due to the delivery of the aqueous polyether solution PE. The droplets D that flow out of the specific well 111S are discharged from the outlet 117 and recovered in the droplet recovery unit 170 via the conduit 190.

[0182] In step S139, the monitoring unit 120 ends the monitoring.

[0183] In step S140, the monitoring results are output via the output unit 150. This completes the series of operation flows.

[0184] A compound library may be used as the sample containing the second component.

[0185] The droplet control device of the second embodiment, having the above-described configuration, can recover droplets containing a desired component even from wells with a very small volume, such as on the order of picoliters or femtoliters. Conventionally, it has been difficult to selectively recover specific droplets from wells with such very small volumes. The droplet control device of the second embodiment utilizes a photoisomerizable compound, allowing the selective recovery of specific droplets even from wells with such very small volumes.

[0186] [Method for Preparing Droplets] A second aspect of the present invention is a method for preparing droplets. In one embodiment, the method for preparing droplets uses a droplet container including a plurality of wells and a flow path communicating with the plurality of wells. In one embodiment, the method for preparing droplets includes the following (a), (b), and (c1) in this order, or the following (b), (a), and (c2) in this order: (a) a step of delivering an aqueous solution containing a polysaccharide to the plurality of wells via the flow path; (b) a step of delivering an aqueous solution containing a polyether to the plurality of wells via the flow path; (c1) a step of delivering an aqueous solution containing the polyether and a photoisomerizable compound to the plurality of wells via the flow path; and (c2) a step of delivering an aqueous solution containing the polysaccharide and a photoisomerizable compound to the plurality of wells via the flow path.

[0187] As the droplet container, the container 110 or the container 110' in the droplet control device 100 of the second embodiment can be used.

[0188] In the case of a method including steps (a), (b), and (c1) in this order, steps (a), (b), and (c1) can be performed in the same manner as steps S101, S102, and S103, respectively.

[0189] In the case of a method including steps (b), (a), and (c2) in this order, steps (b), (a), and (c2) can be performed in the same manner as steps S111, S112, and S113, respectively.

[0190] In one embodiment, the droplet preparation method may further include the following step (d) after step (c1) or (c2): (d) irradiating wells other than the specific well with light of a wavelength that photoisomerizes the photoisomerizable compound. Step (d) can be performed in the same manner as step S105 or step S115.

[0191] In one embodiment, the droplet preparation method may include the following steps (e1) or (e2) after step (d): (e1) a step of delivering an aqueous solution containing a polyether to a plurality of wells via a flow path and recovering droplets from specific wells; and (e2) a step of delivering an aqueous solution containing a polysaccharide to a plurality of wells via a flow path and recovering droplets from specific wells. Step (e1) can be performed in the same manner as step S107 above. Step (e2) can be performed in the same manner as step S117 above.

[0192] The droplet preparation method of this embodiment allows stable droplets to be formed and maintained in wells with microvolumes on the order of picoliters or femtoliters, and by controlling the position of light irradiation, specific droplets can be selectively recovered from any well.

[0193] [Device for Separating a Target Substance] A third aspect of the present disclosure is a device for separating a target substance from a sample prepared in an aqueous medium containing a photoisomerizable compound. In one embodiment, the device for separating a target substance includes a container having a plurality of compartments for accommodating the sample, a light irradiation unit for irradiating the sample accommodated in the compartments with light, and a monitoring unit for monitoring the sample accommodated in the compartments. The light irradiation unit irradiates the sample with light of a wavelength that photoisomerizes the photoisomerizable compound. The sample containing the photoisomerizable compound undergoes a sol-gel transition due to the photoisomerization of the photoisomerizable compound.

[0194] (Target substance) The target substance is not particularly limited. Examples of the target substance include organic synthetic compounds such as low molecular weight compounds, fluorescent dyes, radical generators, and synthetic polymer compounds; biomolecules such as amino acids, nucleotides, sugars, vitamins, peptides, proteins, and nucleic acids; inorganic compounds such as inorganic nanoparticles, inorganic anions, inorganic cations, inorganic salts, and aggregates thereof; cells such as animal cells, plant cells, fungi, algae cells, and protozoan cells; microstructures such as phages, viruses, and molecular aggregates; and cell aggregates such as organoids and spheroids.

[0195] (Sample) A sample can be prepared by mixing a photoisomerizable compound with a test composition containing a target substance. Examples of test compositions containing a target substance include, but are not limited to, body fluids (blood, plasma, serum, saliva, tissue fluid, lymph, urine, pleural effusion, ascites, cerebrospinal fluid, bile, pancreatic juice, amniotic fluid, nasal discharge, tears, sweat, etc.), cell culture medium, cell culture supernatant, cell lysate, cell lysate, and compound library.

[0196] (Photoisomerizable compound) Examples of the photoisomerizable compound include those similar to those described above. In one embodiment, the photoisomerizable compound is a compound that causes the sample to undergo a sol-gel transition upon photoisomerization of the photoisomerizable compound. Specific examples of the photoisomerizable compound include the compound (P) described below.

[0197] (Aqueous Medium) Examples of the aqueous medium include the same as those described above.

[0198] (Configuration Example of Target Substance Separation Apparatus) Figure 23 is a schematic diagram showing a configuration example of a target substance separation apparatus. Separation apparatus 200 includes a container 210, a monitoring unit 220, a light irradiation unit 230, an output unit 250, an input unit 251, a communication unit 252, a memory 253, waste liquid storage units 268, 212, a sample preparation unit 270, and a target substance recovery unit 211, as well as a processor 240 for controlling the operation of each of these units. Separation apparatus 200 also includes a test composition storage unit 261, a photoisomerizable compound storage unit 263, a detection reagent storage unit 266, and an aqueous medium storage unit 267 as storage units for various components.

[0199] In the separation device 200, components for sample preparation are supplied from the test composition storage section 261 and the photoisomerizable compound storage section 263 to the sample preparation unit 270. The sample preparation unit 270 mixes and stirs these components to prepare a sample. The prepared sample is transferred to the container 210. The container 210 has multiple compartments for storing the sample. The sample transferred to the container 210 is distributed among the compartments of the container 210. In one embodiment, the sample transferred to the container 210 is in a sol state.

[0200] The samples contained in each compartment of the container 210 are irradiated with light from the light irradiation unit 30. As a result, the photoisomerizable compound contained in the sample is photoisomerized. The sample containing the photoisomerized photoisomerizable compound gels.

[0201] <<Container>> The container 210 is not particularly limited as long as it has multiple compartments. The multiple compartments may be multiple wells. Examples of the container 210 include those similar to the container 110 of the droplet control device of the second embodiment. Examples of the container 210 include microwell plates (e.g., 96-well microplates), nanowell plates, etc.

[0202] <<Monitoring Unit>> The monitoring unit 220 is for monitoring the state of the sample contained in each compartment of the container 210. The monitoring unit 220 is not particularly limited as long as it can observe the state of the sample contained in each compartment. As the monitoring unit 220, one equipped with the above-mentioned microscope unit can be used.

[0203] <<Light Irradiation Unit>> The light irradiation unit 230 is for irradiating light onto the samples stored in each compartment within the container 210. As a light source for the light irradiation unit 230, a light source similar to that of the light irradiation unit 130 in the droplet control device of the second embodiment can be used.

[0204] <<Sample Preparation Unit>> The sample preparation unit 270 is for preparing a sample. The sample preparation unit 270 prepares a sample by mixing and stirring the test composition and the aqueous photoisomerizable compound solution supplied from the test composition storage section 261 and the photoisomerizable compound storage section 263, respectively.

[0205] The sample preparation unit 270 includes a mixing tank for mixing and stirring the components, and a stirring mechanism 271 for stirring the liquid in the mixing tank. The stirring mechanism 271 is not particularly limited, but examples thereof include a mechanism for rotating a stirrer having a rod-like, plate-like, or propeller-like shape, a mechanism for rotating a stirring blade fixed in the mixing tank, and a mechanism for rotating the mixing tank itself.

[0206] The sample preparation unit 270 may include a temperature controller for controlling the temperature in the mixing tank. Examples of the temperature controller include a heater, a cooler, and a thermostat.

[0207] A waste liquid storage section 268 is connected to the sample preparation unit 270. The waste liquid storage section 268 stores waste liquid used to wash the inside of the mixing tank after, for example, the sample prepared in the mixing tank has been transferred to the container 210. The inside of the mixing tank is washed, for example, with an aqueous medium supplied from an aqueous medium storage section 267. The connection and disconnection between the sample preparation unit 270 and the waste liquid storage section 268 is switched by a valve B5.

[0208] <<Each Component Storage Section>> The test composition storage section 261 is for storing the test composition. The test composition stored in the test composition storage section 261 is preferably in liquid form. The test composition may be a solution or a suspension. The test composition storage section 261 is connected to the sample preparation unit 270 by a conduit 290. The connection and disconnection between the test composition storage section 261 and the sample preparation unit 270 is switched by a valve B1.

[0209] The photoisomerizable compound storage section 263 is used to store an aqueous photoisomerizable compound solution. The concentration of the aqueous photoisomerizable compound solution stored in the photoisomerizable compound storage section 263 is preferably 0.3% by mass or more. In the sample prepared in the sample preparation unit 270, the concentration of the photoisomerizable compound may be 0.1 to 10% by mass relative to the total mass of the sample. The concentration of the aqueous photoisomerizable compound solution stored in the photoisomerizable compound storage section 263 may be any concentration that allows the concentration of the photoisomerizable compound to be adjusted within the aforementioned concentration range when the sample is prepared. The aqueous photoisomerizable compound solution stored in the photoisomerizable compound storage section 263 may be a concentrated solution, and in this case, it may be diluted by supplying an aqueous medium from the aqueous medium storage section 267. The photoisomerizable compound storage section 263 is connected to the sample preparation unit 270 by a conduit 290. The connection and disconnection between the photoisomerizable compound storage section 263 and the sample preparation unit 270 is switched by valves B1 and B2. The photoisomerizable compound stored in the photoisomerizable compound storage section 263 is preferably an isomer that produces a sol state. For example, in the case of a photoisomerizable compound whose trans isomer is in a gel state and whose cis isomer is in a sol state, the cis isomer is preferably stored in the photoisomerizable compound storage section 263. In the case of compound (P) described below, it is preferable that the cis isomer of compound (P) be stored in the photoisomerizable compound storage section 263.

[0210] The detection reagent storage section 266 is for storing an aqueous detection reagent solution. The detection reagent is a reagent that can specifically detect a target substance. The detection reagent is supplied to the sample preparation unit 270 during sample preparation. Examples of the detection reagent include a reagent containing a substance that specifically binds to the target substance (antibody, aptamer, etc.). The detection reagent may be a substance that specifically binds to the target substance to which a labeling substance (such as a fluorescent dye) is bound. Examples of the fluorescent dye include the same as those described above.

[0211] The detection reagent storage section 266 is connected to the sample preparation unit 270 by a conduit 290. The connection and disconnection between the detection reagent storage section 266 and the sample preparation unit 270 is switched by valves B1, B2, and B3.

[0212] Aqueous medium storage section 267 is for storing an aqueous medium. The aqueous medium is used for dilution during sample preparation and for cleaning the mixing tank of sample preparation unit 270 after sample preparation. Aqueous medium storage section 267 is connected to sample preparation unit 270 by conduit 290. The connection and disconnection between aqueous medium storage section 267 and sample preparation unit 270 is switched by valves B1, B2, B3, and B4.

[0213] The output unit 250, input unit 251, and communication unit 252 can be the same as those in the droplet control device.

[0214] <<Processor>> The processor 240 controls the operation of the separation device 200. The operation of each of the above units of the separation device 200 is controlled by the processor 240. The processor 240 realizes these controls by, for example, executing a program stored in the memory 253. The hardware configuration of the processor 240 can be the same as that in the droplet control device.

[0215] The processor 240 has a container control unit 241, a light irradiation unit control unit 242, a sample preparation unit control unit 243, a monitoring unit control unit 244, an output unit control unit 245, an input unit control unit 246, and a communication unit control unit 247, and has the function of controlling the operation of each of the above units.

[0216] The container control unit 241 has a function of controlling the container 210. The container control unit 241 may have a function of controlling the temperature controller 210t to control the temperature of the internal space of the container 210.

[0217] The light irradiation unit control unit 242 has a function of controlling the light irradiation unit 230. The light irradiation unit control unit 242 may have a function of controlling at least one operation selected from the group consisting of the timing of starting light irradiation, the light irradiation time, the wavelength of the irradiation light, and the light irradiation position in the light irradiation unit 230, for example. The light irradiation unit control unit 242 may have a function of controlling a photomask. For example, the light irradiation unit control unit 242 may have a function of selecting a section to block the irradiation light based on the monitoring information of the monitoring unit 220 and placing a photomask at the position of the specific section. Alternatively, the light irradiation unit control unit 242 may have a function of controlling a laser device included in the light irradiation unit 230 to irradiate sections other than the specific section with laser light.

[0218] The sample preparation unit controller 243 has a function of controlling the sample preparation unit 270. The sample preparation unit controller 243 may have a function of controlling the supply of each component from the test composition storage section 261, the photoisomerizable compound storage section 263, the detection reagent storage section 266, and the aqueous medium storage section 267 to the sample preparation unit 270. For example, the sample preparation unit controller 243 may control the supply of these components to the sample preparation unit 270 by controlling valves B1 to B4. The sample preparation unit controller 243 may have a function of controlling the stirring mechanism 271 of the sample preparation unit 270. For example, the sample preparation unit controller 243 may have a function of controlling the operation timing, stirring time, stirring strength, etc. of the stirring mechanism 271. The sample preparation unit controller 243 may have a function of controlling the transfer of the sample from the sample preparation unit 270 to the container 210. For example, the sample preparation unit controller 243 may control the transfer of the sample from the sample preparation unit 270 to the container 210 by controlling pump P. The sample preparation unit control unit 243 may have a function to control the cleaning of the mixing chamber of the sample preparation unit 270. For example, the sample preparation unit control unit 243 may have a function to control the timing of supply of aqueous medium from the aqueous medium storage unit 267 to the sample preparation unit 270, and the discharge of waste liquid to the waste liquid storage unit 268. For example, the sample preparation unit control unit 243 may control the cleaning of the sample preparation unit 270 by controlling the valves B1 to B5. The sample preparation unit control unit 243 may have a function to control the temperature in the mixing chamber of the sample preparation unit 270.

[0219] The monitoring unit control unit 244 has a function of controlling the monitoring unit 220. The monitoring unit control unit 244 may have a function of controlling at least one operation of the monitoring unit 220 selected from the group consisting of the timing of starting monitoring, the monitoring time, and the monitoring position, for example.

[0220] The output unit control unit 245, input unit control unit 246, and communication unit control unit 247 are the same as those in the droplet control device.

[0221] (Example of Operation of Demultiplexer) FIG. 24 is a flowchart showing an example of operation of the demultiplexer 200.

[0222] In step S201, the test composition and photoisomerizable compound are supplied from the test composition storage section 261 and the photoisomerizable compound storage section 263 to the container 210, respectively, to the sample preparation unit 270. If necessary, a detection reagent may be supplied from the detection reagent storage section 266 to the sample preparation unit 270. Alternatively, if necessary, an aqueous medium may be supplied from the aqueous medium storage section 267 to the sample preparation unit 270. The amount of each component supplied may be set in advance, or may be set by the user via the input unit 251. The supply of each component to the sample preparation unit 270 may be performed by a pump (not shown) or the like.

[0223] In step S202, the sample is prepared by mixing and stirring the mixture of the components supplied to the mixing tank of the sample preparation unit 270. The stirring time may be set in advance, or may be set by the user via the input unit 251. Alternatively, the stirring time may be manually stopped by the user via the input unit 251.

[0224] In step S203, the sample prepared in step S202 is stored in each compartment of the container 210. The sample is transferred from the sample preparation unit 270 to the container 210 by controlling the pump P. The entire amount of the sample may be transferred to the container 210, or a portion may be transferred to the container 210 and the remainder may be stored in the waste liquid storage section 268.

[0225] In step S204, the monitoring unit 220 starts monitoring the samples contained in each compartment of the container 210. The monitoring status may be displayed on a display or the like that is the output unit 250. Furthermore, monitoring images or videos may be recorded as needed in the memory 253 or an external storage device.

[0226] In step S205, the light irradiation unit 230 irradiates light onto the compartment of the container 210 that does not contain the target substance. The wavelength of the light irradiated at this time is a wavelength that photoisomerizes the photoisomerizable compound. The light irradiation time may be a time sufficient to photoisomerize the photoisomerizable compound. The light irradiation time may be, for example, the same time as that of the droplet control device described above.

[0227] The compartments to be irradiated with light are selected based on monitoring information from the monitoring unit 220. For example, the monitoring unit 220 detects compartments containing the target substance based on a signal from a detection reagent or the like, and selects compartments that do not contain the target substance as compartments to be irradiated with light. In the compartments irradiated with light, the sample gels due to photoisomerization of the photoisomerizable compound.

[0228] In step S206, the light irradiation unit 230 ends the light irradiation.

[0229] In step S207, the aqueous medium is delivered from the aqueous medium storage section 267 to the container 210 via the sample preparation unit 270. The delivery of the aqueous medium is performed by the sample preparation unit control section 243 controlling valves B1 to B4, and the container control section 241 controlling pump P and valves B6 and B7. At this time, valves B6 and B7 are controlled so that the container 210 is connected to the target substance recovery unit 211. The compartments not containing the target substance were not irradiated with light in step S206, and the sample is in a sol state. Therefore, the target substance can be discharged from the compartment by delivering the aqueous medium. On the other hand, the sample in the compartment irradiated with light in step S206 is in a gel state, and therefore the sample does not flow out of the compartment even when the aqueous medium is delivered. Therefore, the target substance can be selectively recovered. The delivery of the aqueous medium continues until the recovery of the target substance from the compartment containing the target substance is completed.

[0230] In step S208, the monitoring unit 220 ends the monitoring.

[0231] In step S209, the monitoring results are output via the output unit 250. This completes the series of operation flows.

[0232] In step S205, the compartment containing the target substance may be irradiated with light. In this case, in step S207, an aqueous medium is sent to container 210, thereby discharging substances other than the target substance from the compartment not irradiated with light into waste liquid storage section 268. At this time, valves B6 and B7 are controlled so that container 210 is connected to waste liquid storage section 268. The target substance can be recovered by being fixed in container 210 due to gelation of the photoisomerizable compound.

[0233] [Method for Separating a Target Substance] A fourth aspect of the present invention is a method for separating a target substance from a sample prepared in an aqueous medium containing a photoisomerizable compound. In one embodiment, the method for separating a target substance includes the following steps (A) to (C) in this order: (A) distributing the sample prepared in a sol-state aqueous medium containing the photoisomerizable compound into a plurality of compartments; (B) irradiating (i) a compartment containing the target substance or (ii) a compartment not containing the target substance with light of a wavelength that photoisomerizes the photoisomerizable compound, thereby gelling the sample contained in the compartment irradiated with the light; and (C) removing the sample from the compartment not irradiated with the light.

[0234] The container 210 in the separation device 200 can be used as the container containing multiple compartments.

[0235] Steps (A), (B), and (C) can be performed in the same manner as step S203, steps S204 to S206, and step S207, respectively.

[0236] [Composition] A third aspect of the present invention is a composition contained in a light-shielding container. In one embodiment, the composition includes a sugar, a polyether, a photoisomerizable compound, a first droplet encapsulating a first component, and a second droplet encapsulating a second component.

[0237] In one embodiment, the first component and the second component are components that react with each other. Examples of the first component and the second component include components that react with each other to cure. For example, the first component may be an isocyanate compound having an isocyanate group, and the second component may be a compound having a functional group (e.g., an epoxy group, an amino group, a carboxy group, etc.) that is reactive with the isocyanate group.

[0238] The first droplet and the second droplet may be produced by the same method as that described in the droplet control device of the first embodiment.

[0239] In one embodiment, the photoisomerizable compound contained in the composition is an isomer that forms a gel state. For example, when a trans form forms a gel state, the composition contains the trans form of the photoisomerizable compound. When the photoisomerizable compound is compound (P), the composition contains the trans form of compound (P).

[0240] The light-shielding container is not particularly limited as long as it is a container made of a material that blocks light. Examples of the light-shielding container include a metal tube and a resin tube containing a light-shielding agent. The light-shielding container preferably has an opening and closing mechanism such as a lid.

[0241] The composition of the present embodiment can be used, for example, as a two-component adhesive, a two-component coating, a two-component resin, etc. In one embodiment, the composition may be a multi-component formulation contained in a resealable, light-shielding container.

[0242] <Compound (P)> Examples of photoisomerizable compounds include compounds (compound (P)) containing a peptide structure represented by the following general formula (p1) or (p2): XaXb(XaXc) ｎ XaXdXa ... (p1) XaXd (XaXc) ｎXaXbXa (p2) [In the formula, Xa represents an amino acid residue having a hydrophilic side chain; Xb represents an amino acid residue having a side chain represented by the following general formula (b1); Xc and Xd each independently represent an amino acid residue having a hydrophobic side chain or a glycine residue; and n represents an integer of 1 to 3. Multiple Xa's may be the same or different. When n is 2 or 3, multiple Xc's may be the same or different.]

[0243] [In the formula, Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent; Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. * represents a bond.]

[0244] A "peptide structure" refers to a structure in which amino acids are bonded via amide bonds to form a polymer. A compound containing a peptide structure may be a peptide, or may be a compound containing a chemical structure other than a peptide. When referring to a specific peptide structure, the specific peptide structure may be a partial structure contained in one molecule of peptide.

[0245] In the formula (p1) or (p2), Xa represents an amino acid residue having a hydrophilic side chain. The hydrophilic side chain is an amino acid side chain having a hydrophilic group. Xa is preferably an amino acid residue derived from an α-amino acid. Specific examples of Xa include arginine residue, asparagine residue, aspartic acid residue, glutamic acid residue, glutamine residue, lysine residue, serine residue, threonine residue, cysteine ​​residue, histidine residue, and ornithine residue. In the formula (p1) or (p2), multiple Xa may be the same or different.

[0246] In the formula (p1) or (p2), Xb represents an amino acid residue having a side chain represented by the formula (b1) above (hereinafter also referred to as "side chain (b1)"). In formula (b1), * represents a bond bonded to a carbon atom constituting the main chain of the amino acid polymer. Xb is preferably an amino acid residue derived from an α-amino acid. When Xb is an amino acid residue derived from an α-amino acid, the α-amino acid from which Xb is derived is represented by the following formula (Xb-1):

[0247] [In the formula, Ar １ , and Ar ２ represents Ar in the formula (b1). １ , and Ar ２ are the same as

[0248] In formula (b1), Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent. １ The aryl group in the formula (I) preferably has 6 to 18 carbon atoms, more preferably 6 to 15 carbon atoms, and even more preferably 6 to 12 carbon atoms. The aryl group may be a monocyclic group or a polycyclic group. The number of aromatic hydrocarbon rings contained in the aryl group is preferably 1 to 3, more preferably 1 or 2, and even more preferably 1. Specific examples of the aryl group include a phenyl group, a naphthyl group, an anthrenyl group, and a phenanthrenyl group.

[0249] Ar １The heteroaryl group in the above formula preferably has 1 to 18 carbon atoms, more preferably 2 to 15 carbon atoms, and even more preferably 3 to 12 carbon atoms. Examples of heteroatoms contained in the aromatic heterocycle of the heteroaryl group include an oxygen atom, a nitrogen atom, and a sulfur atom. The heteroaryl group may be a monocyclic group or a polycyclic group. The heteroaryl group may be a monocyclic group of an aromatic heterocycle, a fused ring group of an aromatic hydrocarbon ring and an aromatic heterocycle, or a fused ring group of aromatic heterocycles. The total number of aromatic heterocycles and aromatic hydrocarbon rings contained in the heteroaryl group is preferably 1 to 3, more preferably 1 or 2, and even more preferably 1. Examples of aromatic heterocycles contained in heteroaryl groups include nitrogen-containing aromatic heterocycles (pyrrole ring, pyridine ring, azepine ring, azocine ring, azonine ring, imidazole ring, pyrazole ring, imidazoline ring, triazole ring, tetrazole ring, pyridazine ring, pyrimidine ring, pyrazine ring, etc.); fused rings of nitrogen-containing aromatic heterocycles and benzene rings (indole ring, isoindole ring, benzimidazole ring, purine ring, benzotriazole ring, quinoline ring, isoquinoline ring, quinazoline ring, quinoxaline ring, cinnoline ring, etc.); sulfur-containing aromatic heterocycles (thiol ring, thiepine ring, thionine ring, etc.); aromatic heterocycles containing nitrogen and sulfur atoms (thiazole ring, thiazine ring, etc.); oxygen-containing aromatic heterocycles (oxole ring, oxepin ring, oxonine ring, etc.); and aromatic heterocycles containing nitrogen and oxygen atoms (oxazole ring).

[0250] Ar １The aryl group or heteroaryl group in the formula (I) may or may not have a substituent. The substituent is not particularly limited, but a hydrophobic group is preferred. Examples of the substituent include an alkyl group, a halogenated alkyl group, an alkoxy group, a halogenated alkoxy group, and a halogen atom. The alkyl group and alkoxy group as the substituent preferably have 1 to 12 carbon atoms, more preferably 1 to 10 carbon atoms, still more preferably 1 to 6 carbon atoms, and particularly preferably 1 to 3 carbon atoms. Examples of the halogen atom in the halogenated alkyl group and halogenated alkoxy group, and the halogen atom as the substituent, include a fluorine atom, a bromine atom, and an iodine atom, with a fluorine atom being preferred.

[0251] Ar １ is preferably an aryl group which may have a substituent, more preferably a phenyl group or naphthyl group which may have a substituent, still more preferably a phenyl group which may have a substituent, and particularly preferably a perfluorophenyl group (pentafluorophenyl group) or a phenyl group.

[0252] In formula (b1), Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. ２ The arylene group in the formula (I) preferably has 6 to 18 carbon atoms, more preferably 6 to 15 carbon atoms, and even more preferably 6 to 12 carbon atoms. The arylene group may be a monocyclic group or a polycyclic group. The number of aromatic hydrocarbon rings contained in the arylene group is preferably 1 to 3, more preferably 1 or 2, and even more preferably 1. Specific examples of the arylene group include a phenylene group, a naphthylene group, an anthracenediyl group, and a phenanthrenediyl group.

[0253] Ar ２The heteroarylene group in the above formula preferably has 1 to 18 carbon atoms, more preferably 2 to 15 carbon atoms, and even more preferably 3 to 12 carbon atoms. Examples of heteroatoms contained in the aromatic heterocycle of the heteroarylene group include an oxygen atom, a nitrogen atom, and a sulfur atom. The heteroarylene group may be a monocyclic group or a polycyclic group. The heteroarylene group may be a monocyclic group of an aromatic heterocycle, a fused ring group of an aromatic hydrocarbon ring and an aromatic heterocycle, or a fused ring group of aromatic heterocycles. The total number of aromatic heterocycles and aromatic hydrocarbon rings contained in the heteroaryl group is preferably 1 to 3, more preferably 1 or 2, and even more preferably 1. Examples of aromatic heterocycles contained in the heteroarylene group include those mentioned above in Ar １ Examples of the heteroaryl group include the same as the heteroaryl group in the above.

[0254] Ar ２ The arylene group or heteroarylene group in the formula (I) may or may not have a substituent. The substituent is not particularly limited, but a hydrophobic group is preferred. Examples of the substituent include the above-mentioned Ar １ The substituents are the same as those in the above.

[0255] Ar ２ is preferably an arylene group which may have a substituent, more preferably a phenylene group or naphthylene group which may have a substituent, and even more preferably a phenylene group.

[0256] The side chain (b1) is preferably a side chain represented by the following general formula (b1-1).

[0257] [In the formula, m1 and m2 each independently represent an integer of 0 to 3; R １ and R ２ each independently represents a substituent; k1 and k2 each independently represent an integer of 0 or more, as far as the valence allows. When k1 is an integer of 2 or more, two or more R １ When k2 is an integer of 2 or more, two or more R ２ may be the same or different. * represents a bond.]

[0258] In the formula (b1-1), m1 and m2 each independently represent an integer of 0 to 3. m1 and m2 are preferably 0 to 2, more preferably 0 or 1, and even more preferably 0.

[0259] In the formula (b1-1), R １ and R ２ Each of the groups independently represents a substituent. The substituent is not particularly limited, but a hydrophobic group is preferable. Specific examples of the substituent include Ar １ The same substituents as those in R １ and R ２ As the atom, a halogen atom is preferable, and a fluorine atom is more preferable.

[0260] When m1 is 0, k1 is an integer from 0 to 5. When m1 is 1, k1 is an integer from 0 to 7. When m1 is 2, k1 is an integer from 0 to 9. When m1 is 3, k1 is an integer from 0 to 11. m1 is preferably 0, and k1 is preferably 0 or 5. When m2 is 0, k2 is an integer from 0 to 4. When m2 is 1, k2 is an integer from 0 to 6. When m2 is 2, k2 is an integer from 0 to 8. When m2 is 3, k2 is an integer from 0 to 10. m2 is preferably 0, and k1 is preferably 0.

[0261] Specific examples of the side chain (b1) are shown below, but are not limited to these. In the following formula, * represents a bond bonded to a carbon atom constituting the main chain of the amino acid polymer. The following specific examples show trans isomers, but cis isomers are also acceptable.

[0262]

[0263] In the formula (p1) or (p2), Xc and Xd represent an amino acid residue having a hydrophobic side chain or a glycine residue. The amino acid residue having a hydrophobic side chain is preferably an amino acid residue derived from an α-amino acid. Specific examples of the amino acid residue having a hydrophobic side chain include an alanine residue, a valine residue, a leucine residue, an isoleucine residue, a methionine residue, a proline residue, a phenylalanine residue, a tryptophan residue, and a tyrosine residue.

[0264] Xc is preferably an alanine residue or a glycine residue from the viewpoint of the solubility of compound (P) in water. When n in formula (p1) or (p2) is 2 or 3, a plurality of Xc's may be the same or different.

[0265] Xd is preferably an amino acid residue having a hydrophobic side chain having 3 or more carbon atoms. The hydrophobic side chain having 3 or more carbon atoms preferably has 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and even more preferably 3 to 8 carbon atoms. Specific examples of Xd include valine residue, leucine residue, isoleucine residue, methionine residue, proline residue, phenylalanine residue, tryptophan residue, and tyrosine residue.

[0266] In the formula (p1) or (p2), n represents an integer of 1 to 3, preferably 1 or 2, and more preferably 1.

[0267] The peptide structure (p) is preferably a peptide structure represented by the following general formula (p1-1) or (p2-1).

[0268] [In the formula, Ra represents a hydrophilic side chain; Rb represents a side chain represented by the general formula (b1) above; Rc and Rd each independently represent a hydrophobic side chain or a hydrogen atom; and n represents an integer of 1 to 3. Multiple Ra's may be the same or different. When n is 2 or 3, multiple Rc's may be the same or different. * represents a bond.]

[0269] In formula (p1-1) or (p2-1), Ra represents a hydrophilic side chain. Examples of the hydrophilic side chain include an aliphatic hydrocarbon group having a hydrophilic group. Examples of the hydrophilic group include an amino group (preferably a primary amino group), a carboxy group, a hydroxyl group (preferably an alcoholic hydroxyl group), and a thiol group. The aliphatic hydrocarbon group may be a saturated aliphatic hydrocarbon group or an unsaturated aliphatic hydrocarbon group, with a saturated aliphatic hydrocarbon group being preferred. The aliphatic hydrocarbon group may be linear, branched, or cyclic, with a linear or branched chain being preferred, and a linear chain being preferred. The aliphatic hydrocarbon group preferably has 1 to 6 carbon atoms, more preferably 1 to 4 carbon atoms, and even more preferably 1 to 3 carbon atoms.

[0270] The hydrophilic side chain in Ra includes the side chain of a hydrophilic amino acid. Specific examples of hydrophilic amino acids include arginine, asparagine, aspartic acid, glutamine, glutamic acid, lysine, serine, threonine, cysteine, histidine, and ornithine. Specific examples of Ra are shown below, but are not limited to these. In the formula (p1-1) or (p2-1), multiple Ra may be the same or different. In the formula below, * represents a bond bonded to a carbon atom constituting the main chain of the amino acid polymer in the formula (p1-1) or (p2-1). The symbol in parentheses represents the amino acid from which it is derived.

[0271]

[0272] In the formula (p1-1) or (p2-1), Rb represents a side chain represented by the formula (b1). Rb is preferably a side chain represented by the formula (b1-1), and more preferably a side chain represented by any one of the formulas (b1-1-1) to (b1-1-7).

[0273] In the formula (p1-1) or (p2-1), Rc and Rd represent a hydrophobic side chain or a hydrogen atom. Examples of the hydrophobic side chain include hydrocarbon groups having 1 or more carbon atoms, which may have a substituent. The hydrocarbon group preferably has 1 to 12 carbon atoms, more preferably 1 to 10 carbon atoms, and even more preferably 1 to 8 carbon atoms. The hydrocarbon group may be an aromatic hydrocarbon group or an aliphatic hydrocarbon group. The aliphatic hydrocarbon group may be a saturated aliphatic hydrocarbon group or an unsaturated aliphatic hydrocarbon group, with saturated aliphatic hydrocarbon groups being preferred. The aliphatic hydrocarbon group may be linear, branched, or cyclic. The hydrocarbon group may have a substituent as long as it does not become hydrophilic. Examples of the hydrophobic side chain in Rc and Rd include the side chain of a hydrophobic amino acid. Examples of the hydrophobic amino acid include the same as those listed above for Xc and Xd.

[0274] From the viewpoint of the solubility of compound (P) in water, Rc is preferably a hydrogen atom or a methyl group. When n in formula (p1-1) or (p2-1) is 2 or 3, multiple Rc's may be the same or different.

[0275] Rd is preferably a hydrophobic side chain having 3 or more carbon atoms. Examples of hydrophobic side chains having 3 or more carbon atoms include hydrocarbon groups having 3 or more carbon atoms, which may have a substituent. The hydrocarbon group preferably has 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and even more preferably 3 to 8 carbon atoms. The hydrocarbon group may be an aromatic hydrocarbon group or an aliphatic hydrocarbon group. The aliphatic hydrocarbon group may be a saturated aliphatic hydrocarbon group or an unsaturated aliphatic hydrocarbon group, with saturated aliphatic hydrocarbon groups being preferred. The aliphatic hydrocarbon group may be linear, branched, or cyclic. The hydrocarbon group may have a substituent as long as it does not become hydrophilic. Examples of substituents that the hydrocarbon group in Rd may have include a secondary amino group, a tertiary amino group, a sulfide group, a phenolic hydroxyl group, etc.

[0276] Examples of hydrophobic side chains having 3 or more carbon atoms include side chains of hydrophobic amino acids having 3 or more carbon atoms. Specific examples of such hydrophobic amino acids include valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, and tyrosine. Specific examples of Rd are shown below, but are not limited to these. When n in formula (p1-1) or (p2-1) is 2 or 3, multiple Rds may be the same or different. In the following formulas, * represents a bond bonded to a carbon atom constituting the main chain of the amino acid polymer in formula (p1-1) or (p2-1). The symbol in parentheses represents the amino acid from which it is derived.

[0277]

[0278] When Rd is a proline side chain, Rd has the following structure together with the carbon atom to which Rd is bonded in formula (p1-1) or formula (p2-1) and the NH group adjacent to the carbon atom:

[0279]

[0280] In the formula (p1-1) or (p2-1), n ​​represents an integer of 1 to 3, preferably 1 or 2, and more preferably 1.

[0281] In the formula (p1-1) or (p2-1), examples of combinations of Ra, Rb, Rc, and Rd include those in which Ra is a side chain of an amino acid selected from the group consisting of arginine, asparagine, aspartic acid, glutamine, glutamic acid, lysine, serine, threonine, cysteine, histidine, and ornithine; Rb is a side chain represented by the formula (b1-1); Rc is a hydrogen atom or a methyl group; and Rd is a side chain of an amino acid selected from the group consisting of valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, and tyrosine. In these combinations, n in the formula (p1-1) or (p2-1) is preferably 1 or 2, and more preferably 1. In the formula (p1-1) or (p2-1), a combination of Ra, Rb, Rc, and Rd is preferably a combination in which Ra is a side chain of arginine or aspartic acid, Rb is a side chain represented by the formula (b1-1), Rc is a hydrogen atom or a methyl group, and Rd is a side chain of methionine. In the above combination, n in the formula (p1-1) or (p2-1) is preferably 1 or 2, and more preferably 1.

[0282] Specific examples of the peptide structure (p) are shown below, but are not limited to these. In the specific examples below, the side chain (b1) is in a trans form, but the side chain (b1) may be in a cis form. * indicates a bond binding to an adjacent atom.

[0283]

[0284]

[0285] In compound (P), the structure other than the peptide structure (p) is not particularly limited. Compound (P) may be, for example, a peptide containing a peptide structure (p). In this case, the number of amino acid residues contained in compound (P) is not particularly limited, and examples include 40 or less, 30 or less, 25 or less, 20 or less, 15 or less, or 10 or less. The number of amino acid residues includes the number of amino acid residues contained in peptide structure (p). When compound (P) is a peptide containing peptide structure (p), the type of amino acid residues in the region other than peptide structure (p) is not particularly limited. In compound (P), the portion other than peptide structure (p) does not have to be a peptide. For example, an organic group having 1 to 30 carbon atoms may be bonded to either or both of the two terminals of peptide structure (p). Compound (P) may be a compound represented by the following general formula (p1-1') or (p2-1'):

[0286] [In the formula, R Ａ and R Ｂ each independently represents a monovalent group. Ra, Rb, Rc, Rd, and n are the same as Ra, Rb, Rc, Rd, and n in formula (p1-1) or (p2-1), respectively.

[0287] R in general formula (p1-1′) or (p2-1′) Ａ and R Ｂ The monovalent group in R is not particularly limited. Ａ and R Ｂ Examples of the monovalent group in include an acetyl group, an amino group, a hydroxy group, a hydrogen atom, a halogen atom, and a monovalent organic group. Examples of the monovalent organic group include a peptide consisting of 1 to 20 amino acid residues and a monovalent organic group other than a peptide. Examples of the monovalent organic group other than a peptide include a hydrocarbon group which may have a substituent. Specific examples include, but are not limited to, an alkyl group, a polyoxyethylene group, a polyoxypropylene group, and a phenyl group.

[0288] Specific examples of the compound (P) are shown below, but are not limited to these. In the specific examples below, the side chain (b1) is in a trans form, but the side chain (b1) may be in a cis form.

[0289]

[0290]

[0291] Compound (P) can be produced by known peptide synthesis methods such as solid-phase peptide synthesis. Commercially available amino acids can be used as raw materials for compound (P).

[0292] The present invention will be described below with reference to examples, but the present invention is not limited to the following examples.

[0293] [Synthesis of Photoisomerizable Compound (Compound (P1))] Compound (P1) represented by the above formula (P1) was synthesized as a photoisomerizable compound by solid-phase peptide synthesis. Phenylazophenylalanine used in peptide synthesis was purchased from Tokyo Chemical Industry Co., Ltd., and other amino acids (arginine, aspartic acid, alanine, methionine) were purchased from Watanabe Chemical Industry Co., Ltd. The synthesized compound (P1) was analyzed using matrix-assisted laser desorption / ionization (MALDI). A peak (observed value 1096.65) attributable to the molecular weight of the protonated product of compound (P1) (theoretical value 1096.55) was observed. This result confirmed that compound (P1) was obtained.

[0294] Compound (P1) is a compound that photoisomerizes from a trans isomer to a cis isomer when irradiated with light of 350 nm, and from a cis isomer to a trans isomer when irradiated with light of 450 nm.

[0295]

[0296] Example 1 Preparation of Droplets The following samples A and B were prepared.

[0297] Sample A: Polyethylene glycol (35 kDa; PEG35k, 5 wt%), dextran (550 kDa; DEX550k, 5 wt%), compound (P1) (1 wt%), Escherichia coli, fluorescein-di-β-D-galactopyranoside (FDG, 60 μM), and isopropyl-β-D-thiogalactopyranoside (IPTG, 170 μM) were dispersed in water.

[0298] Sample B: Polyethylene glycol (35 kDa; PEG35k, 5 wt %), dextran (550 kDa; DEX550k, 5 wt %), compound (P1) (1 wt %), and M13 phage virus were dispersed in water.

[0299] Sample A was stirred by vortexing for 30 seconds to prepare a droplet (Droplet A). Droplet A was considered to contain E. coli, FDG, and IPTG (see FIG. 17).

[0300] Sample B was stirred by vortexing for 30 seconds to prepare a droplet (droplet B). Droplet B was considered to contain encapsulated M13 phage virus (see FIG. 17).

[0301] Thereafter, 2 μL of the dispersion of droplet A and 2 μL of the dispersion of droplet B were collected, placed in one microtube, and mixed by pipetting to prepare a droplet dispersion containing droplets A and B (see FIG. 17 ).

[0302] <Light Irradiation> 4 μL of the droplet dispersion prepared as described above was taken and dropped onto a plate to prepare a droplet dispersion plate. Two droplet dispersion plates were prepared. One plate was irradiated with light (UNA) at 360 to 370 nm for 5 minutes. The other plate was not irradiated with light. Thereafter, the plates were incubated at 37°C, 5% CO ２ Two plates were incubated for one day under the conditions.

[0303] <Observation of Phage Infection into E. coli> After the incubation, the two plates were observed under a fluorescence microscope (Olympus IX-73). The results are shown in Figure 18.

[0304] In the plate that was not irradiated with light, almost no green fluorescence was observed. This was thought to be because the M13 phage did not infect the E. coli and the FDG was not decomposed by the β-galactosidase contained in the M13 phage. The faint signal observed was thought to be due to scattering.

[0305] On the other hand, strong green fluorescence was observed in the plate that had been irradiated with light. This was thought to be due to the fusion of droplets A and B upon light irradiation, resulting in infection of the E. coli with the M13 phage. Specifically, it was thought that the β-D-galactopyranoside of FDG was decomposed by the β-galactosidase contained in the M13 phage, resulting in the emission of fluorescence (see Figure 17). Compound (P1) is isomerized from the trans form to the cis form upon irradiation with light of 360 to 370 nm. The trans form of compound (P1) is fibrous, and the droplet dispersion is gel-like. Therefore, in the presence of the trans form of compound (P1), droplet fusion hardly occurs. On the other hand, the cis form of compound (P1) loses its fibrous structure, resulting in the droplet dispersion becoming sol-like. Therefore, in the presence of the cis form of compound (P1), droplet fusion occurs. It was believed that upon irradiation with light at 360 to 370 nm, compound (P1) was isomerized from a trans form to a cis form, resulting in the fusion of droplets A and B.

[0306] Example 2 Femtoliter Chamber Array Device (FRAD) The FRAD was fabricated by photolithographic microfabrication according to Ueno et al. (Ueno, H. et al., Protein Sci 2021, 30(8), 1628-1639.) The flow cell was assembled using the FRAD, a top glass with inlet and outlet holes, and double-sided tape (~80 μm) as a spacer. The top glass of the flow cell chamber was precoated with CYTOP 809M to prevent nonspecific binding of biomolecules. The well volume of the fabricated FRAD was 6.3 fL.

[0307] <Sample Preparation> The following solutions were prepared. DEX aqueous solution: DEX550k was dissolved in water to a concentration of 10 wt %. PEG aqueous solution: PEG35k was dissolved in water to a concentration of 5 wt %. P1 / PEG aqueous solution: PEG35k and compound (P1) were dissolved in water to a concentration of 5 wt % (PEG) and 1 wt % (compound (P1)), respectively.

[0308] <Preparation of droplets> Figure 19 shows an outline of the procedure in Example 2. First, a DEX aqueous solution was pumped into the flow path in the FRAD ((1) in Figure 19). The pumping rate was 200 µL / min. Next, a PEG aqueous solution was pumped into the flow path in the FRAD ((2) in Figure 19). The pumping rate was 200 µL / min. This resulted in the formation of DEX droplets in the wells of the FRAD. Next, a P1 / PEG aqueous solution was pumped into the flow path in the FRAD ((3) in Figure 19). The pumping rate was 200 µL / min.

[0309] <Recovery of Droplets> Next, without light irradiation, an aqueous PEG solution was pumped into the channel in the FRAD ((4) in FIG. 19). The pumping speed was set to 200 μL / min.

[0310] Photographs of the FRAD before and after the injection of the aqueous PEG solution are shown in Figure 20. It was confirmed that many of the droplets present in the well before the injection of the PEG solution were shed by the injection of the aqueous PEG solution. This result confirmed that droplets could be recovered from femtoliter-sized wells by injecting the aqueous PEG solution without irradiating the well with light.

[0311] [Example 3] <Preparation of Samples> The following solutions were prepared. DEX aqueous solution: DEX550k was dissolved in water to a concentration of 10 wt%. PEG aqueous solution: PEG35k was dissolved in water to a concentration of 5 wt%. P1 / PEG aqueous solution: PEG35k and compound (P1) were dissolved in water to a concentration of 5 wt% (PEG) and 1 wt% (compound (P1)), respectively. 3T3 cell / PEG aqueous solution: 1x10 ６ 3T3 cells were suspended at 1000 cells / mL.

[0312] <Preparation of droplets> Figure 21 shows an outline of the procedure in Example 3. First, a DEX aqueous solution was pumped into the flow path in the FRAD ((1) in Figure 21). The pumping rate was 200 µL / min. Next, a 3T3 cell / PEG aqueous solution was pumped into the flow path in the FRAD ((2) in Figure 21). The pumping rate was 200 µL / min. This resulted in the formation of a DEX droplet containing 3T3 cells in the well of the FRAD. Next, a P1 / PEG aqueous solution was pumped into the flow path in the FRAD ((3) in Figure 21). The pumping rate was 200 µL / min. Two FRADs in which droplets were prepared using the above procedure were prepared.

[0313] <Comparison of droplet recovery rates with and without light irradiation> After (3) in Figure 21, an aqueous PEG solution was pumped through the flow path in one FRAD without light irradiation ((4) in Figure 21). The pumping rate was 200 μL / min. After (3) in Figure 21, the other FRAD was irradiated with light (350 m, 10 minutes) ((5) in Figure 21). Then, an aqueous PEG solution was pumped through the flow path in the FRAD ((6) in Figure 21). The pumping rate was 200 μL / min.

[0314] Figure 22 shows photographs of the FRAD after the PEG aqueous solution was pumped with or without light irradiation. When the PEG aqueous solution was pumped with or without light irradiation, 27 cells remained in the FRAD. On the other hand, when the PEG aqueous solution was pumped with light irradiation, 46 cells remained in the FRAD. These results confirmed that irradiation with light of a wavelength that photoisomerizes compound (P1) increases the retention rate of droplets containing 3T3 cells.

[0315] According to the present invention, a droplet control device and a droplet production method capable of controlling liquid-liquid phase separation droplets are provided, and a composition capable of controlling the initiation of a reaction of multiple components is also provided.

[0316] Although the preferred embodiments of the present invention have been described above, the present invention is not limited to these embodiments. Addition, omission, substitution, and other modifications of the configuration are possible within the scope of the spirit of the present invention. The present invention is not limited by the above description, but is limited only by the scope of the appended claims.

[0317] 1,100 Droplet control device 10, 110, 210 Container 10t, 110t, 210t Temperature controller 20, 120, 120 Monitoring unit 21 Microscope unit 22 Objective lens 23 Dichroic mirror 24 Imaging lens 25 Pupil projection lens 26 Control mirror 27 Light source 28 Image sensor 30, 130, 230 Light irradiation unit 40, 140, 240 Processor 41, 141, 241 Container control unit 42, 142, 242 Light irradiation unit control unit 43 Droplet preparation unit control unit 44, 144, 244 Monitoring unit control unit 45, 145, 245 Output unit control unit 46, 146, 246 Input unit control unit 47, 147, 247 Communication unit control unit 50, 150, 250 Output unit 51, 151, 251 Input unit 52, 152, 252 Communication unit 53, 153, 253 Memory 61, 161 Polysaccharide storage section 62, 162 Polyether storage section 63, 163, 263 Photoisomerizable compound storage section 64, 164 First component storage section 65, 165 Second component storage section 66, 166, 266 Detection reagent storage section 67, 167, 267 Aqueous medium storage section 68, 168, 212, 268 Waste liquid storage section 70 Droplet preparation unit 71, 271 Stirring mechanism 160 Liquid delivery unit 170 Droplet recovery unit 110a, 110a' Ceiling substrate 110b, 110b' Bottom substrate 110c, 110c' Side wall 111, 111' Well 112, 112' Partition wall 115, 115' Flow path 116, 116' Supply port 117, 117' Discharge port 90, 190, 290 Conduit 211 Target substance recovery unit 243 Sample preparation unit control section 261 Test composition storage section 270 Sample preparation unit

Claims

1. A droplet control device for controlling droplets made from polysaccharides and polyethers, comprising: a light irradiation unit for irradiating light; and a monitoring unit for monitoring the state of the droplets, wherein an aqueous medium containing the droplets further contains a photoisomerizable compound, and the light irradiation unit irradiates the aqueous medium with light of a wavelength that photoisomerizes the photoisomerizable compound.

2. The droplet controller of claim 1, further comprising a processor for controlling operation of said droplet controller.

3. The droplet control device according to claim 1 or 2, further comprising a container for containing the aqueous medium.

4. The droplet control device according to claim 2, wherein the processor controls the operation of the light irradiation unit, and the operation of the light irradiation unit controlled by the processor is at least one selected from the group consisting of timing to start light irradiation, light irradiation time, wavelength of the irradiated light, and light irradiation position.

5. The droplet control device according to claim 2, wherein the processor controls the operation of the monitoring unit, and the operation of the monitoring unit controlled by the processor is at least one selected from the group consisting of a monitoring start timing, a monitoring time, and a monitoring position.

6. The droplet control device according to claim 1 or 2, further comprising an output unit for outputting a monitoring result by the monitoring unit.

7. A droplet control device according to claim 1 or 2, wherein the monitoring unit comprises a microscope unit for magnifying and observing the droplet.

8. The droplet control device of claim 1 or 2, wherein the droplets include: a first droplet including a first component; and a second droplet including a second component, the first component being a component that interacts with the second component.

9. The droplet control device of claim 8, further comprising: a first component storage section for storing the first component; a second component storage section for storing the second component; a polysaccharide storage section for storing a polysaccharide; a polyether storage section for storing a polyether; and a photoisomerizable compound storage section for storing the photoisomerizable compound.

10. The droplet control device according to claim 9, further comprising a droplet preparation unit, the droplet preparation unit being controlled to: prepare the first droplet by mixing an aqueous solution containing the first component supplied from the first component storage unit, the polysaccharide supplied from the polysaccharide storage unit, the polyether supplied from the polyether storage unit, and the photoisomerization compound supplied from the photoisomerization compound storage unit; and prepare the second droplet by mixing the second component supplied from the second component storage unit, the polysaccharide supplied from the polysaccharide storage unit, the polyether supplied from the polyether storage unit, and the aqueous solution containing the photoisomerization compound supplied from the photoisomerization compound storage unit.

11. The droplet control device of claim 10, further comprising a detection reagent storage section for supplying a detection reagent, the detection reagent storage section being controlled to supply the detection reagent to the droplet production unit at either or both of the timings when the first droplet is produced and when the second droplet is produced.

12. The droplet control device according to claim 10, wherein the droplet production unit includes a stirring mechanism.

13. The droplet control device of claim 8, wherein the monitoring unit is controlled to monitor an interaction between the first component and the second component within a third droplet produced by fusing the first droplet and the second droplet.

14. The droplet control device according to claim 8, wherein the monitoring unit is controlled to perform monitoring based on a point in time when the light irradiation unit starts irradiating the aqueous medium with light of a wavelength at which the photoisomerizable compound is photoisomerized.

15. The droplet control device of claim 8, wherein the first component is a cell, the second component is a virus, and the interaction is infection of the cell by the virus.

16. The droplet control device of claim 3, wherein the container comprises a plurality of wells and a channel communicating with the plurality of wells.

17. The droplet control device of claim 16, wherein the volume of the well is 1 picoliter or less.

18. The droplet control device of claim 16, further comprising a liquid delivery unit that delivers liquid to the plurality of wells via the flow paths.

19. The droplet control device according to claim 18, wherein the liquid delivery unit comprises: a polysaccharide storage section for supplying a polysaccharide; a polyether storage section for supplying a polyether; and a photoisomerizable compound storage section for supplying the photoisomerizable compound.

20. The droplet control device of claim 19, wherein the liquid delivery unit is controlled to deliver, in the order of: (i) an aqueous solution containing the polysaccharide to the plurality of wells; (ii) an aqueous solution containing the polyether to the plurality of wells; and (iii) an aqueous solution containing the polyether and the photoisomerizable compound to the plurality of wells.

21. The droplet control device of claim 20, wherein the monitoring unit is controlled to begin monitoring after (iii).

22. The droplet control device according to claim 21, wherein the light irradiation unit is controlled to irradiate wells other than a specific well with light of a wavelength at which the photoisomerizable compound photoisomerizes, based on the monitoring result of the monitoring unit.

23. The droplet control device of claim 22, further comprising a droplet recovery unit for recovering said droplet from said particular well.

24. The droplet control device according to claim 1 or 2, wherein the photoisomerizable compound is a compound containing a peptide structure represented by the following general formula (p1) or (p2): XaXb(XaXc) ｎ XaXdXa ... (p1) XaXd (XaXc) ｎ XaXbXa ... (p2) [In the formula, Xa represents an amino acid residue having a hydrophilic side chain; Xb represents an amino acid residue having a side chain represented by the following general formula (b1); Xc and Xd each independently represent an amino acid residue having a hydrophobic side chain or a glycine residue; and n represents an integer of 1 to 3. The multiple Xas may be the same or different. When n is 2 or 3, the multiple Xcs may be the same or different.] [In the formula, Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent; Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. * represents a bond.

25. An apparatus for separating a target substance from a sample prepared in an aqueous medium containing a photoisomerizable compound, comprising: a container having a plurality of compartments for containing the sample; a light irradiation unit for irradiating the sample contained in the compartments with light; and a monitoring unit for monitoring the sample contained in the compartments, wherein the light irradiation unit irradiates the sample with light of a wavelength that photoisomerizes the photoisomerizable compound, and the sample containing the photoisomerizable compound undergoes a sol-gel transition due to the photoisomerization of the photoisomerizable compound.

26. A method for preparing droplets using a droplet container having a plurality of wells and flow paths communicating with the plurality of wells, comprising the following (a), (b), and (c1) in this order, or the following (b), (a), and (c2) in this order: (a) a step of delivering an aqueous solution containing a polysaccharide to the plurality of wells via the flow paths; (b) a step of delivering an aqueous solution containing a polyether to the plurality of wells via the flow paths; (c1) a step of delivering an aqueous solution containing the polyether and a photoisomerizable compound to the plurality of wells via the flow paths; (c2) a step of delivering an aqueous solution containing the polysaccharide and a photoisomerizable compound to the plurality of wells via the flow paths.

27. The method according to claim 26, further comprising, after step (c1) or (c2), a step (d) of irradiating wells other than the specific well with light having a wavelength that photoisomerizes the photoisomerizable compound.

28. The method according to claim 27, further comprising, after step (d), (e1) delivering an aqueous solution containing a polyether to the plurality of wells via the flow path and recovering droplets from the particular wells, or (e2) delivering an aqueous solution containing a polysaccharide to the plurality of wells via the flow path and recovering droplets from the particular wells.

29. A method for separating a target substance from a sample prepared in an aqueous medium containing a photoisomerizable compound, comprising: (A) a step of distributing the sample prepared in an aqueous medium in a sol state containing the photoisomerizable compound into a plurality of compartments; (B) a step of irradiating (i) the compartment containing the target substance or (ii) the compartment not containing the target substance with light of a wavelength that photoisomerizes the photoisomerizable compound, thereby gelling the sample contained in the compartment irradiated with the light; and (C) a step of removing the sample from the compartment not irradiated with the light.

30. The method according to any one of claims 26 to 29, wherein the photoisomerizable compound is a compound containing a peptide structure represented by the following general formula (p1) or (p2): XaXb(XaXc) ｎ XaXdXa ... (p1) XaXd (XaXc) ｎ XaXbXa ... (p2) [In the formula, Xa represents an amino acid residue having a hydrophilic side chain; Xb represents an amino acid residue having a side chain represented by the following general formula (b1); Xc and Xd each independently represent an amino acid residue having a hydrophobic side chain or a glycine residue; and n represents an integer of 1 to 3. The multiple Xas may be the same or different. When n is 2 or 3, the multiple Xcs may be the same or different.] [In the formula, Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent; Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. * represents a bond.

31. A composition contained in a light-tight container, the composition comprising: a polysaccharide, a polyether, a photoisomerizable compound, a first droplet having a first component encapsulated therein, and a second droplet having a second component encapsulated therein.

32. The composition according to claim 31, wherein the photoisomerizable compound is a compound containing a peptide structure represented by the following general formula (p1) or (p2): XaXb(XaXc) ｎ XaXdXa ... (p1) XaXd (XaXc) ｎ XaXbXa ... (p2) [In the formula, Xa represents an amino acid residue having a hydrophilic side chain; Xb represents an amino acid residue having a side chain represented by the following general formula (b1); Xc and Xd each independently represent an amino acid residue having a hydrophobic side chain or a glycine residue; and n represents an integer of 1 to 3. The multiple Xas may be the same or different. When n is 2 or 3, the multiple Xcs may be the same or different.] [In the formula, Ar １ represents an aryl group which may have a substituent or a heteroaryl group which may have a substituent; Ar ２ represents an arylene group which may have a substituent or a heteroarylene group which may have a substituent. * represents a bond.

33. A multiple component formulation comprising: an openable, light-shielding container; and the composition according to claim 31 or 32 contained in said light-shielding container.

Images