Hydrogel flow channel device with sensor

Abstract

The hydrogel channel device (1) with a sensor according to the present invention comprises a solid substrate (10) and a hydrogel laminate (20) located on the solid substrate (10). The solid substrate (10) has, at the interface with the hydrogel laminate (20), an adhesive area that is bonded to the hydrogel laminate (20) and a non-adhesive area that is not bonded to the hydrogel laminate (20). The hydrogel laminate (20) has a swelling gel thin film layer (21) on the solid substrate (10) and a non-swelling gel layer (22) laminated on the swelling gel thin film layer (21). The present invention has, between the solid substrate (10) and the hydrogel laminate (20), a hydrogel channel (30) formed by separating the swelling gel thin film layer (21) in the non-adhesive area, and has a sensor part at the interface between the solid substrate (10) and the hydrogel laminate (20).

Description

[Technical Field]

[0001] The present invention relates to a hydrogel flow channel type device with a sensor. [Background technology]

[0002] In recent years, attempts have been made to mimic biological functions and structures in vitro using microfluidic chips in drug discovery research aimed at treating diseases and in research to elucidate the mechanisms of disease onset. One example of such attempts is the construction of a model system known as a biomimetic system (MPS). Microfluidic chips are widely used in MPS. The substrate used for MPS is made of a material that is suitable for cell culture and is easily moldable. Examples of such materials include glass, polymethyl methacrylate (PMMA), and polydimethylsiloxane (PDMS).

[0003] On the other hand, in MPS, it is necessary to evaluate the effects of oral drugs and understand the dynamics of infectious disease and disease-causing substances when they act on the diseased site. To do this, it is necessary to evaluate the diffusion and effects of drugs and causative substances that reach the site of action in biological tissues from the circulatory system, including blood vessels. The materials used in conventional microfluidic devices have low substance permeability and are not suitable for the above purposes. Therefore, there is a demand for materials suitable for the above purposes.

[0004] To achieve the above objectives, it is known that hydrogels, which have high substance permeability and biocompatibility, are effective. Microfluidic devices using hydrogels are known, for example, to have a three-dimensional structure resulting from the free swelling of the hydrogel in the non-adhesive regions, with adhesive / non-adhesive regions between a solid substrate and a swelling gel arranged in a pattern (see, for example, Patent Document 1). Hydrogel channel-type devices that utilize three-dimensional structures as flow channels are also known (see, for example, Patent Document 2). Furthermore, hydrogel channel-type devices are known to be usable as substance-permeable microfluidic devices for the diffusion of dyes from the channels, cell culture on the hydrogel channel-type device, and drug stimulation of cells (see, for example, Non-Patent Document 1). [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Publication No. 2020-62843 [Patent Document 2] International Publication No. 2021 / 079399 [Non-patent literature]

[0006] [Non-Patent Document 1] “Tough, permeable and biocompatible microfluidic devices formed through the buckling delamination of soft hydrogel films”,Riku Takahashi,et al., The Royal Society of Chemistry 2021,Lab Chip,2021,21,1307-1317 Summary of the Invention [Problem to be solved by the invention]

[0007] When measuring substance permeation through a hydrogel channel-type device, the use of dyes is essential to visualize the diffusion of substances into the device over time. However, when using MPS for pharmacodynamic analysis or to measure the dynamics of disease-causing substances, it is often impossible to visualize the target drug or causative substance (hereinafter referred to as "analyte"). One method for visualizing analytes is to modify the analyte with a labeling agent (e.g., fluorescent dye, nanoparticles). However, this method has drawbacks, such as being cumbersome and the possibility that modification with a labeling agent can change the permeability and diffusion rate of the target substance.

[0008] The present invention has been made in consideration of the above circumstances, and aims to provide a hydrogel channel-type device with a sensor that can create a tissue-like structure that simulates the biological environment on a chip outside the body, and that can measure molecular diffusion into and out of the structure via a blood vessel-like tubular structure. [Means for solving the problem]

[0009] One aspect of the present invention is a hydrogel channel device with a sensor, comprising: a solid substrate; and a hydrogel laminate located on the solid substrate; the solid substrate having, at its interface with the hydrogel laminate, an adhesive region where the solid substrate is adhered to the hydrogel laminate and a non-adhesive region where the hydrogel laminate is not adhered to the hydrogel laminate; the hydrogel laminate having a swellable gel thin film layer on the solid substrate and a non-swellable gel layer laminated on the swellable gel thin film layer; a hydrogel channel between the solid substrate and the hydrogel laminate, where the swellable gel thin film layer is separated by the non-adhesive region; and a sensor unit at the interface between the solid substrate and the hydrogel laminate. [Effects of the Invention]

[0010] The present invention makes it possible to create tissue-like structures that mimic the biological environment on a chip outside of a living body, and to provide a hydrogel flow channel device with a sensor that can measure molecular diffusion into and out of the structure via a blood vessel-like tubular structure. [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 is a perspective view showing a schematic configuration of a hydrogel channel-type device with a sensor according to one embodiment of the present invention. [Figure 2] FIG. 1 is a perspective view showing a schematic configuration of a solid substrate and a hydrogel laminate that constitute a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. [Figure 3] FIG. 1 is a plan view showing a schematic configuration of a solid substrate constituting a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. [Figure 4] FIG. 1 is a side view showing a schematic configuration of a solid substrate constituting a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. [Figure 5] FIG. 5 is an enlarged view of region α shown in FIG. 4, illustrating a schematic configuration of a solid substrate that constitutes a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. [Figure 6] FIG. 5 is an enlarged view of region α shown in FIG. 4, illustrating a schematic configuration of a solid substrate that constitutes a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. [Figure 7] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution in a hydrogel channel-type device with an SPR sensor and detection by the SPR sensor. [Figure 8] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution in a hydrogel channel-type device with an SPR sensor and detection by the SPR sensor. [Figure 9] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution in a hydrogel channel-type device with an SPR sensor and detection by the SPR sensor. [Figure 10] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution in a hydrogel channel-type device with an SPR sensor and detection by the SPR sensor. [Figure 11]FIG. 10 is a diagram showing the relationship between the diffusion position of an analyte solution in a hydrogel laminate and the diffusion time of the analyte solution in the hydrogel laminate. [Figure 12] FIG. 10 is a graph showing the relationship between the diffusion time of an analyte solution into a hydrogel laminate and the intensity of the light absorption spectrum on the detection surface of the sensor part. [Figure 13] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution through a hydrogel flow channel device with a graphene / DNA aptamer sensor and detection by the graphene / DNA aptamer sensor. [Figure 14] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution through a hydrogel flow channel device with a graphene / DNA aptamer sensor and detection by the graphene / DNA aptamer sensor. [Figure 15] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution through a hydrogel flow channel device with a graphene / DNA aptamer sensor and detection by the graphene / DNA aptamer sensor. [Figure 16] FIG. 1 is a side view illustrating the concept of diffusion of an analyte solution through a hydrogel flow channel device with a graphene / DNA aptamer sensor and detection by the graphene / DNA aptamer sensor. [Figure 17] FIG. 10 is a diagram showing the relationship between the diffusion position of an analyte solution relative to a hydrogel laminate and the fluorescence intensity of a fluorescent dye at the end of a DNA aptamer. [Figure 18] FIG. 10 is a plan view showing a schematic configuration of a modified example of a solid substrate constituting a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. [Figure 19] FIG. 10 is a side view showing a schematic configuration of a modified example of a solid substrate constituting a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. [Figure 20] FIG. 1 shows the results of measuring the diffusion of a rhodamine solution in a hydrogel channel-type device equipped with an SPR sensor using an SPR sensor in an example. DETAILED DESCRIPTION OF THE INVENTION

[0012] A sensor-equipped hydrogel channel-type device according to an embodiment of the present invention will be described in detail below with reference to the drawings. Note that the drawings used in the following description may show characteristic portions enlarged for ease of understanding, and the dimensional proportions of the components may not be the same as those in reality. Furthermore, the materials, dimensions, etc. exemplified in the following description are merely examples, and the present invention is not limited thereto. Appropriate modifications may be made within the scope of the present invention.

[0013] [Hydrogel flow channel device with sensor] FIG. 1 is a perspective view showing a schematic configuration of a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. FIG. 2 is a perspective view showing a schematic configuration of a solid substrate and a hydrogel laminate that constitute a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. FIG. 3 is a plan view showing a schematic configuration of a solid substrate that constitutes a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. FIG. 4 is a side view showing a schematic configuration of a solid substrate that constitutes a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention. FIGS. 5 and 6 are enlarged views of region α shown in FIG. 4, showing a schematic configuration of a solid substrate that constitutes a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention.

[0014] As shown in FIGS. 1 to 4, the sensor-equipped hydrogel channel device 1 of this embodiment includes a solid substrate 10 and a hydrogel laminate 20 located on the solid substrate 10. As shown in FIG. 2, the solid substrate 10 has, at its interface with the hydrogel laminate 20, an adhesive region 11 that is adhered to the hydrogel laminate 20 and a non-adhesive region 12 that is not adhered to the hydrogel laminate 20. Here, the interface of the solid substrate 10 with the hydrogel laminate 20 is one surface 10a of the solid substrate 10 (the upper surface shown in FIGS. 1 to 4). As shown in FIGS. 1 and 2, the hydrogel laminate 20 includes a swellable gel thin film layer 21 on one surface 10a of the solid substrate 10 and a non-swellable gel layer 22 laminated on the swellable gel thin film layer 21. The swellable gel thin film layer 21 is adhered to the adhesive region 11 but not to the non-adhesive region 12. The sensor-equipped hydrogel channel device 1 has a hydrogel channel 30 between a solid substrate 10 and a hydrogel laminate 20, with a swellable gel thin film layer 21 spaced apart by a non-adhesive region 12. As shown in Figures 3 and 4, the sensor-equipped hydrogel channel device 1 has a sensor section 40 at the interface between the solid substrate 10 and the hydrogel laminate 20. As shown in Figure 1, the sensor-equipped hydrogel channel device 1 preferably includes a liquid supply tube 50 connected to the hydrogel channel 30.

[0015] "Solid substrate" A solid substrate 10 supports a hydrogel laminate 20 . The solid substrate 10 is not particularly limited as long as it does not impair the effects of the present invention, and examples thereof include a substrate made of an inorganic material such as glass or silicon, and a plastic film made of an organic material such as polysilicone or polyurethane. The solid substrate 10 may have one surface 10a covered with a thin film made of a metal or inorganic oxide having any desired function, or a thin film of any desired shape made of an organic material having any desired function.

[0016] The adhesive region 11 has a first adhesive region 11A and a second adhesive region 11B with a non-adhesive region 12 interposed therebetween. The adhesive region 11 and the non-adhesive region 12 are strip-shaped in plan view, with one side longer. In FIG. 1 , the X direction is the longitudinal direction (length) of the adhesive region 11 and the non-adhesive region 12, and the Y direction is the lateral direction (width) of the adhesive region 11 and the non-adhesive region 12. The longitudinal direction of the solid substrate 10 and the hydrogel laminate 20 is the same as the longitudinal direction of the adhesive region 11 and the non-adhesive region 12. The lateral direction of the solid substrate 10 and the hydrogel laminate 20 is the same as the lateral direction of the adhesive region 11 and the non-adhesive region 12. The non-adhesive region 12 is arranged in a strip shape inside the hydrogel channel 30. The adhesive region 11 is arranged on both sides of the non-adhesive region 12 in the extension direction.

[0017] (Sensor section) The sensor unit 40 is provided on the adhesive region 11 and the non-adhesive region 12. The sensor unit 40 has a plurality of unit sensor unit rows 42, each consisting of two or more unit sensor units 41, lined up in the longitudinal direction of the adhesive region 11 and the non-adhesive region 12. Here, an example is shown in which the sensor unit 40 has a first unit sensor unit row 42A, a second unit sensor unit row 42B, a third unit sensor unit row 42C, a fourth unit sensor unit row 42D, and a fifth unit sensor unit row 42E, each consisting of unit sensor units 41. The first unit sensor unit row 41A, the second unit sensor unit row 42B, the third unit sensor unit row 42C, the fourth unit sensor unit row 42D, and the fifth unit sensor unit row 42E are lined up at a distance from each other in the width direction of the adhesive region 11 and the non-adhesive region 12. The sensor unit 40 is composed of a plurality of unit sensor units 41 provided at a distance from each other on one surface 10a of the solid substrate 10. In other words, the sensor unit 40 is a collection of a plurality of unit sensor units 41 provided on one surface 10a of the solid substrate 10. The unit sensor units 41 constitute a first unit sensor unit row 41A, a second unit sensor unit row 42B, a third unit sensor unit row 42C, a fourth unit sensor unit row 42D, and a fifth unit sensor unit row 42E. In addition, the unit sensor units 41 are present discontinuously in the length direction of one surface 10a of the solid substrate 10 in a plan view. Note that in this embodiment, "discontinuously" means that the unit sensor units 41 have an island-like structure, and there are portions on one surface 10a of the solid substrate 10 where the solid substrate 10 is exposed.

[0018] In the adhesion region 11, a hydrogel laminate 20 (swellable gel thin film layer 21) is adhered to one surface 10a of the solid substrate 10. Therefore, the sensor section 40 is covered with the hydrogel laminate 20 (swellable gel thin film layer 21). On the other hand, in the non-adhesion region 12, the hydrogel laminate 20 (swellable gel thin film layer 21) is not adhered to one surface 10a of the solid substrate 10. Therefore, the sensor section 40 is not covered with the hydrogel laminate 20 (swellable gel thin film layer 21).

[0019] As shown in FIG. 5, in the sensor unit 40, each unit sensor element 41 has a detection surface 43 and a probe 44. The detection surface 43 is the outermost surface (top surface) of the unit sensor element 41. The detection surface 43 detects changes in the concentration of an analyte in the vicinity of the sensor unit 40. The probe 44 is provided on the detection surface 43 so as to protrude in the thickness direction of the solid substrate 10. The probe 44 specifically binds to the analyte. Note that each unit sensor element 41 may have one type of probe or two or more types of probes. That is, in the sensor unit 40, each unit sensor element 41 may have a probe 45 shown in FIG. 6 that is different from the probe 44, or may have both the probe 44 and the probe 45.

[0020] The types and combinations of the detection surface 43 and probe 44 of the sensor section 40 are not limited as long as they can detect the target analyte. Examples of materials that can be used to form the detection surface 43 include a gold thin film, gold nanoparticles, and graphene.

[0021] Examples of materials constituting the probe 44 include an antibody to which the analyte specifically binds, and a DNA aptamer modified with a fluorescent dye at the end.

[0022] In particular, when the detection surface 43 is made of a gold thin film and the probe 44 is made of an antibody, the sensor unit 40 is used for surface plasmon measurement. Also, when the detection surface 43 is made of graphene and the probe 44 is made of a DNA aptamer, the sensor unit 40 is used for measuring fluorescence intensity.

[0023] (Sacrificial layer) If the adhesion between the sensor part 40 and the swellable gel thin film layer 21 is strong and it is difficult to form a hydrogel flow path 30 by buckling and peeling of the swellable gel thin film layer 21 on the non-adhesive region 12, a sacrificial layer may be formed on the non-adhesive region 12 of the solid substrate 10.

[0024] The sacrificial layer is located between the solid substrate 10 and the swellable gel thin film layer 21 in at least a portion of the non-adhesive region 12. The region of the sacrificial layer becomes a peeling region that peels off upon application of a predetermined stimulant solution stimulus (addition of a chelating agent). The material of the sacrificial layer is not particularly limited as long as it can be dissolved by the predetermined stimulant solution stimulus (addition of a chelating agent). Here, predetermined solution stimuli include chelating agent solution stimulus that binds with calcium ions in calcium alginate in aqueous solution, temperature stimulus, light stimulus, etc. The sacrificial layer is preferably a thin film that can maintain adhesion to one surface 10a of the solid substrate 10 in dry and wet environments (especially physiological environments).

[0025] Materials for the sacrificial layer include calcium alginate, which dissolves when calcium chelating agents such as ethylenediaminetetraacetic acid (EDTA), glycol ether diaminetetraacetic acid, 1,2-bis(o-aminophenoxide)ethane-N,N,N',N'-tetraacetic acid, and citric acid are added; biopolymers such as dextran that can be decomposed by enzymes; gelatin that exhibits a sol-gel transition depending on the temperature; and polymers containing photoisomerizable molecules such as azobenzene and spiropyran that undergo a sol-gel transition when exposed to light.

[0026] The thickness of the sacrificial layer is not particularly limited as long as it is possible to form a hydrogel flow path 30 between solid substrate 10 and swellable gel thin film layer 21 after swelling by a dissolution stimulus.

[0027] "Hydrogel laminate" (Swellable gel thin film layer) The hydrogel laminate 20 has a swellable gel thin film layer 21. The swellable gel thin film layer 21 is made of hydrogel and is laminated on one surface 10a of the solid substrate 10.

[0028] Examples of polymeric materials that make up hydrogels include water-soluble polymers such as polyacrylamide and polyvinyl alcohol, polysaccharides such as chitosan and alginic acid, and proteins such as collagen and albumin. These materials have a three-dimensional network structure and swell by absorbing a solvent to fill most of their volume. Examples of solvents that swell the polymeric materials that make up hydrogels include water.

[0029] Furthermore, a stimuli-responsive polymer material can be used as the polymer material constituting the hydrogel. Here, "stimuli-responsive" refers to the property of the polymer material constituting the hydrogel to change its molecular structure in response to stimuli such as heat, light, electricity, and pH. In a stimuli-responsive hydrogel, the three-dimensional network structure of the polymer material constituting the hydrogel changes in response to a stimulus that changes its molecular structure, thereby changing its degree of swelling. In the following explanation, a hydrogel containing a stimuli-responsive polymer material is sometimes referred to as a "stimuli-responsive hydrogel."

[0030] Examples of such stimulus-responsive polymeric materials include polymeric materials that respond to thermal stimuli, polymeric materials that respond to pH, polymeric materials that respond to light, and polymeric materials that respond to electrical stimuli.

[0031] Examples of polymeric materials that respond to thermal stimuli include poly(N-isopropylacrylamide) and poly(methyl vinyl ether).

[0032] Examples of pH-responsive polymeric materials include polymer electrolytes obtained by polymerizing anionic or cationic monomers.

[0033] Examples of polymeric materials that respond to light include polymeric materials having spiropyran or azobenzene in the molecular skeleton.

[0034] Examples of polymeric materials that respond to electrical stimuli include polypyrrole, polythiophene, and polyaniline.

[0035] The material for forming the swellable gel thin film layer 21 may be a mixture of these polymeric materials to form a hydrogel that responds to multiple stimuli. Furthermore, the material for forming the swellable gel thin film layer 21 may also be a tough hydrogel such as a double network gel, slide-ring gel, Tetra-PEG gel, or nanoclay gel.

[0036] Various known methods can be used to synthesize the polymer material that constitutes the hydrogel. For example, when the polymer material that constitutes the hydrogel is an acrylic polymer material, a three-dimensional network structure may be formed by crosslinking acrylic groups during polymerization of acrylic monomers.

[0037] The type of polymerization reaction when polymerizing the acrylic monomer is not particularly limited, but examples thereof include radical polymerization using a water-soluble photopolymerization initiator. Examples of the water-soluble photopolymerization initiator include 2-oxoglutaric acid, 4'-(2-hydroxyethoxy)-2-hydroxy-2-methylpropiophenone (trade name: Irgacure 2959), lithium phenyl(2,4,6-trimethylbenzoyl)phosphinate (abbreviation: LAP), and 2,2'-azobis[2-methyl-N-(2-hydroxyethyl)propionamide] (trade name: VA-086).

[0038] During radical polymerization, a deoxidizer may be added to the reaction system to prevent polymerization inhibition by oxygen. Examples of the deoxidizer include a combination of glucose and glucose oxidase. Radical polymerization may also be carried out under an inert gas atmosphere such as nitrogen or argon.

[0039] When the polymeric material constituting the hydrogel is a polysaccharide or a protein, the three-dimensional network structure may be formed by physical bonding of the polysaccharide or protein, or by crosslinking the polysaccharide or protein using a crosslinking agent, such as glutaraldehyde.

[0040] The shape of the swellable gel thin film layer 21 is not particularly limited, and various shapes can be selected depending on the mode of use. For example, the swellable gel thin film layer 21 may be in the form of a film, a plate, a block, etc. Among these, the shape of the swellable gel thin film layer 21 is preferably in the form of a film.

[0041] The thickness of the swellable gel thin film layer 21 is not particularly limited, but is preferably a thickness that provides sufficient structural strength to prevent the layer from collapsing under its own weight. For example, when a hydrogel containing polyacrylamide is used as the material for forming the swellable gel thin film layer 21, the thickness of the swellable gel thin film layer 21 is preferably 50 μm to 1000 μm, and more preferably 120 μm to 200 μm.

[0042] The strength of swellable gel thin film layer 21 can be improved by increasing the crosslinking of the polymeric material that constitutes the hydrogel by chemical crosslinking or physical crosslinking, or by increasing the concentration of the polymeric material that constitutes the hydrogel.

[0043] For example, when preparing a hydrogel containing polyacrylamide by polymerizing acrylamide monomers (precursors), the monomer concentration is preferably 0.8 mol / L to 8 mol / L, more preferably 2 mol / L to 4 mol / L.

[0044] Furthermore, when methylenebisacrylamide is used as a chemical crosslinking agent for polymerizing acrylamide monomers, the concentration of the crosslinking agent is preferably 0.01 mol % to 20 mol %, more preferably 0.03 mol % to 1 mol %, based on the monomer.

[0045] Hydrogels can contain various additives. The type of additive is not particularly limited as long as it does not inhibit hydrogel formation. Examples of additives include biomolecules that improve biocompatibility, silver nanoparticles or surfactants that exhibit antibacterial properties, ionic liquids or conductive polymers that increase conductivity, and magnetic nanoparticles that react to magnetic fields. Adding these additives to hydrogels can impart desired functions to the hydrogels.

[0046] (non-swelling gel thin film layer) The hydrogel laminate 20 has a non-swellable gel layer 22 on a swellable gel thin film layer 21. The non-swellable gel layer 22 is made of hydrogel and is laminated on the swellable gel thin film layer 21.

[0047] The non-swelling gel layer 22 is a gel formed after the polymer material constituting the hydrogel has swelled. That is, the non-swelling gel layer 22 is formed when a liquid such as water flows into the network structure of the polymer material, causing the layer to swell. Therefore, the non-swelling gel layer 22 can also be said to be a swollen polymer material.

[0048] The polymer material constituting the non-swelling gel layer 22 has a lower swelling degree than the polymer material constituting the swellable gel thin film layer 21. There are no particular limitations on the swelling degree of the polymer material constituting the non-swelling gel layer 22, as long as it is lower than the swelling degree of the polymer material constituting the swellable gel thin film layer 21. The swelling degree of the polymer material constituting the non-swelling gel layer 22 in one direction is preferably, for example, about 0.8 to 1.2 times the size before swelling.

[0049] Here, the "degree of swelling" can be calculated, for example, by cutting out a disk-shaped sample of an appropriate diameter from the polymeric material constituting the swellable gel thin film layer 21 or the polymeric material constituting the non-swellable gel layer 22 immediately after polymerization, leaving the disk-shaped sample to stand in pure water until no size change occurs, and then using the following formula (1). (Swelling degree) = D / D0(1) In the above formula (1), D is the diameter of the largest part of the sample after being placed in pure water, and D0 is the diameter of the circular sample before being placed in pure water.

[0050] The polymer material constituting non-swelling gel layer 22 may be a hydrogel or a gel other than a hydrogel. When the polymer material constituting non-swelling gel layer 22 is a hydrogel, the polymer material constituting non-swelling gel layer 22 may be the same as or different from the polymer material constituting swelling gel thin film layer 21.

[0051] Examples of polymeric materials constituting non-swelling gel layer 22 include chemically cross-linked gels that are cross-linked by covalent bonds due to a radical polymerization reaction of monomers. Examples of chemically cross-linked gels include polyacrylamide and its derivatives (polydimethylacrylamide, poly-N-isopropylacrylamide, etc.). In this case, methylenebisacrylamide may be used as a cross-linking agent to increase the cross-linking density and keep the swelling degree of the polymeric material constituting non-swelling gel layer 22 within the above-mentioned range.

[0052] The polymer material that constitutes the non-swelling gel layer 22 also includes a physically crosslinked gel that combines a polymer having a positive or negative charge with an ion having a multivalent charge of the opposite polarity.

[0053] Examples of physically cross-linked gels include gels formed by combining sodium alginate solution, a negatively charged polymer, with calcium solutions such as calcium chloride and calcium sulfate. Other examples include gels formed by combining poly(2,2'-disulfo-4,4'-bensidineterephthalamide: PBDT), a water-soluble polyaramid, with various metal polyvalent cations (Ca 2+ ,Fe 2+ ,Al 3+ ,Zr 4+ ,Ti 4+ Examples include physically crosslinked gels in combination with PBDT. TEMPO-oxidized cellulose nanofibers (NIPPON PAPER INDUSTRIES CO., LTD.), which also carry a negative charge, and cellulose nanofibers defibrated by the phosphoric acid esterification method (Oji Holdings Corporation) may also be used instead of PBDT.

[0054] Here, "TEMPO" is an abbreviation for 2,2,6,6-tetramethylpiperidine 1-oxyl (2,2,6,6-tetramethylpiperidine-1-oxyl radical).

[0055] The non-swelling gel layer 22 covers one surface of the swellable gel thin film layer 21 outside the hydrogel channel 30. The non-swelling gel layer 22 covers the surface of the swellable gel thin film layer 21 that is not in contact with the solid substrate 10. In other words, the surface of the swellable gel thin film layer 21 that is covered by the non-swelling gel layer 22 is the surface opposite the surface that is in contact with the solid substrate 10 (i.e., the surface that faces the surface that is in contact with the solid substrate 10).

[0056] The non-swelling gel layer 22 covers the outside of the hydrogel channel 30. Therefore, when an aqueous liquid is poured into the inside of the hydrogel channel 30, the aqueous liquid permeates the swellable gel thin film layer 21, diffuses to the outside of the hydrogel channel 30, and reaches the non-swelling gel layer 22.

[0057] For example, when the non-swelling gel layer 22 is made of a hydrogel, the aqueous liquid that reaches the non-swelling gel layer 22 can diffuse into the non-swelling gel layer 22. Therefore, by placing any object (e.g., cells, cultured tissue) inside the non-swelling gel layer 22 in advance, the aqueous liquid can be selectively supplied to the object in a predetermined region inside the non-swelling gel layer 22.

[0058] For example, when a positively or negatively charged hydrogel is used as the polymer material constituting non-swelling gel layer 22, the function of preventing the diffusion of small molecules with a specific charge can be imparted to hydrogel channel 30. In other words, non-swelling gel layer 22 can be imparted with a shielding function to prevent the diffusion of small molecules with a specific charge from the inside to the outside of hydrogel channel 30.

[0059] Other examples of polymeric materials that can be used to form the non-swelling gel layer 22 include hydrogels that switch between hydrophilic and hydrophobic properties in response to external stimuli; and hydrogels that can change their degree of swelling in response to external stimuli.

[0060] When using a hydrogel whose properties switch between hydrophilic and hydrophobic in response to an external stimulus, if the non-swelling gel layer 22 becomes hydrophilic in response to the external stimulus, the small molecules diffusing within the non-swelling gel layer 22 can be selectively limited to hydrophilic ones. On the other hand, if the non-swelling gel layer 22 becomes hydrophobic in response to the external stimulus, the small molecules diffusing within the non-swelling gel layer 22 can be selectively limited to hydrophobic ones.

[0061] When using a hydrogel that can change its swelling rate (water content) in response to an external stimulus, if the swelling rate of the non-swelling gel layer 22 is relatively high as a result of responding to the external stimulus, the diffusion rate of low molecules diffusing within the non-swelling gel layer 22 becomes relatively slow.

[0062] On the other hand, when the swelling rate of the non-swelling gel layer 22 is relatively low as a result of responding to an external stimulus, the diffusion rate of low molecules diffusing within the non-swelling gel layer 22 becomes relatively fast.

[0063] Additionally, the non-swelling gel layer 22 may have a functional group that exhibits a predetermined response, such as fluorescence, to small molecules diffusing from the hydrogel channel 30. In this case, when small molecules diffuse through the non-swelling gel layer 22, the non-swelling gel layer 22 exhibits a predetermined response, such as fluorescence, and therefore the hydrogel channel-type sensor device 1 can be endowed with the function of a sensor for the diffused small molecules.

[0064] There are no particular limitations on the mechanical strength of the non-swelling gel layer 22. For example, if the non-swelling gel layer 22 is required to have an elastic modulus (up to 1.3 MPa) similar to that of polydimethylsiloxane (PDMS), a double-network gel, which is a composite of a physically cross-linked gel and a chemically cross-linked gel, is preferred as the polymer material constituting the non-swelling gel layer 22. The double-network gel has a strong double-network structure, which further improves the mechanical strength.

[0065] There are no particular limitations on the shape of the non-swelling gel layer 22. However, the thickness of the non-swelling gel layer 22 needs to be greater than the height of the hydrogel channel 30 in order to cover the hydrogel channel 30. The thickness of the non-swelling gel layer 22 may be further increased to ensure sufficient strength at the joint between the liquid delivery tube 50 and the hydrogel laminate 20.

[0066] The polymer material constituting non-swelling gel layer 22 may contain various additives as long as the additives do not cause a significant change in the degree of swelling. By using any additive, any desired function can be imparted to non-swelling gel layer 22.

[0067] The additives in the non-swelling gel layer 22 are not particularly limited as long as they do not inhibit gel formation. Examples include biomolecules that improve biocompatibility, silver nanoparticles and surfactants that exhibit antibacterial properties, ionic liquids and conductive polymers that increase conductivity, magnetic nanoparticles that react to magnetic fields, and proteins that bind to glucose to enhance fluorescence intensity.

[0068] The method for synthesizing the polymer material that constitutes non-swelling gel layer 22 is not particularly limited, as long as it is a method that results in a degree of swelling that is lower than that of the polymer material that constitutes swellable gel thin film layer 21 .

[0069] (Hydrogel channel) The hydrogel channel 30 is formed between the solid substrate 10 and the hydrogel laminate 20. The hydrogel channel 30 is formed by the polymer material constituting the swellable gel thin film layer 21 swelling to separate the polymer material constituting the swellable gel thin film layer 21 from the non-adhesive region 12 of the solid substrate 10.

[0070] Specifically, at the interface between the solid substrate 10 and the swellable gel thin film layer 21, the position of the polymer material constituting the swellable gel thin film layer 21 at which it separates from the solid substrate 10 is controlled by the pattern arrangement of the adhesive regions 11 and non-adhesive regions 12. The polymer material constituting the swellable gel thin film layer 21 swells, and the polymer material constituting the swellable gel thin film layer 21 on the non-adhesive regions 12 selectively separates in the non-adhesive regions 12 of the solid substrate 10, causing buckling deformation of the polymer material constituting the swellable gel thin film layer 21. As a result, a hybrid channel, i.e., a hydrogel channel 30, is formed as a space surrounded by the solid substrate 10 and the swellable gel thin film layer 21.

[0071] The hydrogel channel 30 has a portion of the swellable gel thin film layer 21 that is separated from the solid substrate 10 as a channel surface 30c. The hydrogel channel 30 has a first open end surface 30a and a second open end surface 30b. The channel surface 30c of the hydrogel channel 30 is formed in a strip shape along the extension direction of the non-adhesion region 12 between the first open end surface 30a and the second open end surface 30b. The hydrogel channel 30 formed at the interface between the solid substrate 10 and the swellable gel thin film layer 21 penetrates the non-swellable gel layer 22 from the first end surface 22a to the second end surface 22b.

[0072] Liquid delivery tube The liquid supply tube 50 is fixed by an adhesive to the first opening end face 30a and the second opening end face 30b of the hydrogel channel 30. Specifically, at each of the first opening end face 30a and the second opening end face 30b of the hydrogel channel 30, the liquid supply tube 50 is fixed by an adhesive between the solid substrate 10 and the hydrogel laminate 20. The liquid supply tube 50 is for supplying any fluid into the hydrogel channel 30 .

[0073] The liquid supply tube 50 is not particularly limited as long as it can supply liquid from the outside. The type of the liquid supply tube 50 is not particularly limited. Examples of the liquid supply tube 50 include tubes made of polytetrafluoroethylene (PTFE), tetrafluoroethylene (PFA), polyurethane, polyethylene, silicone, polyimide, etc. The outer diameter of the liquid supply tube 50 is not particularly limited. However, it is desirable that the outer diameter of the liquid supply tube 50 be approximately the same as the height of the hydrogel channel 30.

[0074] The adhesive fixes the liquid supply tube 50 to the hydrogel channel 30. That is, the adhesive fixes the liquid supply tube 50 between the solid substrate 10 and the hydrogel laminate 20.

[0075] In the sensor-equipped hydrogel channel-type device 1, the adhesive is densely filled around the hydrogel channel 30 at the first open end face 30a and the second open end face 30b of the hydrogel channel 30, in the space in contact with the channel face 30c of the hydrogel channel 30. The adhesive is preferably water-resistant and has adhesive properties to the solid substrate 10 and the hydrogel laminate 20. Examples of adhesives include cyanoacrylate adhesives, silicone adhesives, and epoxy adhesives.

[0076] "Mechanism of action" In the hydrogel channel-type device 1 with a sensor described above, when a tissue-like structure consisting of epithelial cells is formed on the upper surface 20a of the hydrogel laminate 20 (upper surface 22c of the non-swelling gel layer 22), the hydrogel channel 30 formed between the solid substrate 10 and the hydrogel laminate 20 can be regarded as circulatory system tubular tissue such as blood vessels and lymph, and the non-swelling gel layer 22 can be regarded as interstitial tissue, thereby simulating the structure of skin or the structure of digestive organs such as the esophagus and intestines.

[0077] In the hydrogel channel-type sensor device 1 described above, vascular endothelial cells can be cultured on the inner wall of the hydrogel channel 30 formed between the solid substrate 10 and the hydrogel laminate 20, fibroblasts can be cultured inside the non-swelling gel layer 22, and epithelial cells can be cultured on the upper surface 20a of the hydrogel laminate 20. Therefore, it is possible to create an MPS with a composition closer to that of living tissue.

[0078] In the above-described sensor-equipped hydrogel channel device 1, the mesh size and hardness of the swellable gel thin film layer 21 and the non-swellable gel layer 22 can be adjusted by changing the hydrogel composition, and by controlling the physical properties of the hydrogel, it is possible to create a diseased tissue model of biological tissue. For example, it is possible to simulate local changes in the physical properties inside biological tissue due to fibrosis or scarring of the biological tissue.

[0079] In the sensor-equipped hydrogel channel type device 1 described above, the sensor section 40 can have one or more types of probes on the detection surface of the unit sensor section 41. Therefore, the dynamics of multiple analytes can be evaluated.

[0080] In the sensor-equipped hydrogel channel type device 1 described above, the sensor section 40 can have one or more types of probes on the detection surface of the unit sensor section 41. Therefore, by simultaneously providing a probe using an optical detection method and a probe using an electrochemical detection method, it is possible to select a detection method suitable for the analyte.

[0081] In the hydrogel channel-type sensor device 1 described above, the sensor unit 40 is provided on the solid substrate 10, and therefore only two-dimensional information can be obtained by the sensor unit 40. However, by assuming that the diffusion of the analyte within the hydrogel laminate 20 is isotropic, it is possible to estimate the spatial distribution of the analyte during the diffusion process. For example, as shown in FIG. 1 , if the height from the inner wall of the hydrogel channel 30 to the upper surface 20a of the hydrogel laminate 20 is d, the arrival time of the analyte at the upper surface 20a of the hydrogel laminate 20 can be estimated by detecting the analyte signal on the solid substrate 10, which is a distance d from the inner wall of the hydrogel channel 30.

[0082] "How to use the sensor-equipped hydrogel flow channel device" <Detection of analyte solution diffusion using a hydrogel flow channel device with a surface plasmon resonance (SPR) sensor> 7 to 10 are side views showing the concept of diffusion of an analyte solution in a hydrogel channel-type device with an SPR sensor and detection by the SPR sensor.

[0083] As shown in Figure 7, one surface 10a of the solid substrate 10 is sputtered with, for example, a thin gold film, forming the detection surface 43 of the sensor unit 40. The radial size w of the detection surface 43 in the hydrogel channel 30 is preferably greater than the height d from the inner wall of the hydrogel channel 30 to the upper surface 20a of the hydrogel laminate 20. Figure 7 shows the state before the analyte solution 100 is pumped into the hydrogel channel 30. t0 in Figure 7 indicates the state before the analyte solution 100 is pumped into the hydrogel channel 30.

[0084] As shown in Figure 8, when the analyte solution 100 is pumped into the hydrogel channel 30, the analyte solution 100 diffuses into the swellable gel thin film layer 21. Also, t1 in Figure 8 indicates the time immediately after the analyte solution 100 is pumped into the hydrogel channel 30.

[0085] 9, the analyte solution 100 also diffuses into the non-swelling gel layer 22, and reaches the upper surface 20a of the hydrogel laminate 20. t2 in FIG. 9 indicates that some time has passed since the analyte solution 100 was delivered into the hydrogel channel 30.

[0086] As time passes, the analyte solution 100 diffuses over a wide area of ​​the non-swelling gel layer 22, as shown in Figure 10. t3 in Figure 10 indicates that a considerable amount of time has passed since the analyte solution 100 was delivered into the hydrogel channel 30.

[0087] The SPR sensor detects the diffusion of the analyte solution 100 by measuring changes in the light absorption spectrum on the detection surface 43 of the sensor unit 40. As a result, as shown in FIG. 11 , the relationship between the diffusion position of the analyte solution 100 relative to the hydrogel laminate 20 and the diffusion time of the analyte solution 100 relative to the hydrogel laminate 20 (the time elapsed after the analyte solution 100 is delivered into the hydrogel channel 30, the time it takes for the analyte solution 100 to reach the upper surface 20a of the hydrogel laminate 20) can be obtained. Furthermore, as shown in FIG. 12 , the relationship between the diffusion time of the analyte solution 100 relative to the hydrogel laminate 20 (the time elapsed after the analyte solution 100 is delivered into the hydrogel channel 30, the time it takes for the analyte solution 100 to reach the upper surface 20a of the hydrogel laminate 20) and the measurement result of the change in the light absorption spectrum on the detection surface 43 of the sensor unit 40 (the intensity of the absorption spectrum) can be obtained.

[0088] The sensor unit 40 may have probes on the detection surface 43. The sensor unit 40 may have one type of probe on the detection surface 43, or may have two or more types of probes provided in an array. The probe is not particularly limited, but examples thereof include antibodies and DNA aptamers that can be easily immobilized on the surface of a thin gold film.

[0089] <Detection of analyte solution diffusion using a hydrogel flow channel device with a graphene / DNA aptamer sensor> 13 to 16 are side views illustrating the concept of diffusion of an analyte solution through a hydrogel fluidic device with a graphene / DNA aptamer sensor and detection by the graphene / DNA aptamer sensor. In FIGS. 13 to 16, white circles indicate cases where the fluorescent dye at the end of the DNA aptamer has high fluorescence intensity, and black circles indicate cases where the fluorescent dye at the end of the DNA aptamer has low fluorescence intensity. Furthermore, the sensor units 40 indicated by A, B, B', C, and C' in FIGS. 13 to 16 correspond to A, B, B', C, and C' in FIG. 17.

[0090] As shown in FIG. 13 , one surface 10a of a solid substrate 10 is immobilized with, for example, graphene or graphene oxide to form the detection surface 43 of the sensor unit 40. The method for immobilizing graphene or graphene oxide on the one surface 10a of the solid substrate 10 is not particularly limited, but examples include a method combining photolithography and oxygen plasma etching, inkjet printing, and the like. The sensor unit 40 has a probe 45 on the detection surface 43. Examples of the probe 45 include a DNA aptamer. The DNA aptamer preferably has a fluorescent dye at the end not immobilized on the detection surface 43, and the end immobilized on the detection surface 43 is modified with a functional group that binds to the detection surface 43. The fluorescent dye at the end of the DNA aptamer is not particularly limited, as long as it changes structure upon binding to an analyte and undergoes fluorescence quenching due to light energy transfer when the fluorescent dye is located near the detection surface 43. Examples of such fluorescent dyes include fluorescein and rhodamine. The functional group for immobilizing the DNA aptamer on the detection surface 43 is not particularly limited as long as it can maintain binding under physiological conditions, and examples thereof include pyrene and amino groups. Figure 13 shows the state before the analyte solution 100 is pumped into the hydrogel channel 30. t0 in Figure 13 indicates the state before the analyte solution 100 is pumped into the hydrogel channel 30.

[0091] 14, when the analyte solution 100 is pumped into the hydrogel channel 30, the analyte solution 100 diffuses into the swelling gel thin film layer 21. Also, t1 in FIG. 14 indicates the time immediately after the analyte solution is pumped into the hydrogel channel 30.

[0092] 15, the analyte solution 100 also diffuses into the non-swelling gel layer 22, and reaches the upper surface 20a of the hydrogel laminate 20. t2 in FIG. 15 indicates that some time has passed since the analyte solution was delivered into the hydrogel channel 30.

[0093] As time passes, the analyte solution 100 diffuses over a wide area of ​​the non-swelling gel layer 22, as shown in Figure 16. Also, t3 in Figure 16 indicates that a considerable amount of time has passed since the analyte solution was delivered into the hydrogel channel 30.

[0094] The graphene / DNA aptamer sensor detects the diffusion of the analyte solution 100 by measuring the fluorescence quenching (fluorescence intensity) of the fluorescent dye at the end of the DNA aptamer. As a result, as shown in Figure 17, the relationship between the diffusion position of the analyte solution 100 relative to the hydrogel laminate 20 and the fluorescence quenching (fluorescence intensity) of the fluorescent dye at the end of the DNA aptamer is obtained. When the analyte solution 100 comes into contact with the fluorescent dye at the end of the DNA aptamer, the fluorescence intensity of the fluorescent dye weakens, allowing the diffusion of the analyte solution 100 to the position of the corresponding DNA aptamer to be detected.

[0095] [Method for manufacturing a hydrogel flow-type device with sensors] 2 and 3, in one example of a method for manufacturing a sensor-equipped hydrogel channel-type device according to one embodiment of the present invention, adhesive regions 11 and non-adhesive regions 12 are formed on one surface 10a of a solid substrate 10 on which a sensor section 40 is provided. The method for forming the adhesive regions 11 and non-adhesive regions 12 is not particularly limited, but examples include a method using photolithography and oxygen plasma etching with a positive photoresist, and a stencil method in which negative molds of the adhesive regions 11 and non-adhesive regions 12 are produced and then subjected to oxygen plasma etching.

[0096] Next, a polymer material constituting the swellable gel thin film layer 21, in other words, a hydrogel precursor solution, is dropped onto one surface 10a of the solid substrate 10, and any desired radical polymerization is performed to form the swellable gel thin film layer 21. There are no particular limitations on the method for bonding the swellable gel thin film layer 21 to the solid substrate 10, but one example is a method in which the one surface 10a of the solid substrate 10 and the swellable gel thin film layer 21 are bonded by covalent bonding. For example, one surface 10a of the solid substrate 10 is modified with 3-(methacryloyloxy)propyltrimethoxysilane (TMSPMA), and a hydrogel precursor solution that gels by radical polymerization is dropped onto the modified surface, and any desired radical polymerization is performed. When a gold thin film is used on the detection surface 43 of the sensor section 40 provided on one surface 10a of the solid substrate 10, the swellable gel thin film layer 21 and the solid substrate 10 may be bonded by modifying the surface of the gold thin film with a compound having a dithiol and an acrylic group, such as bis(2-methacryloyl)oxyethyl disulfide (Bis-thiol).

[0097] Alternatively, a porous thin film may be deposited on one surface 10a of the solid substrate 10, and the swellable gel thin film layer 21 and the solid substrate 10 may be bonded together by mutual penetration.

[0098] Next, a polymeric material constituting the non-swelling gel layer 22, in other words, a precursor solution of a hydrogel, is dropped onto the swelling gel thin film layer 21, and optional radical polymerization is performed to form the non-swelling gel layer 22.

[0099] Next, the polymer material constituting the swellable gel thin film layer 21 is swelled (gelled) and the polymer material constituting the swellable gel thin film layer 21 on the non-adhesive region 12 is selectively separated from the solid substrate 10, thereby causing buckling deformation of the polymer material constituting the swellable gel thin film layer 21 and forming a hydrogel flow path 30 as a space surrounded by the solid substrate 10 and the swellable gel thin film layer 21.

[0100] Next, liquid supply tubes 50 are inserted into both ends of the hydrogel channel 30, and the liquid supply tubes 50 are bonded and fixed between the solid substrate 10 and the hydrogel laminate 20 using an adhesive, thereby obtaining a hydrogel channel device 1 with a sensor.

[0101] [Other embodiments] The present invention is not limited to the above-described embodiment. For example, modifications such as those shown in FIGS. 18 and 19 may be adopted.

[0102] "Variations" 18 and 19, like the above-described solid substrate 10, has, at the interface with the hydrogel laminate 20, an adhesive region 211 that adheres to the hydrogel laminate 20 and a non-adhesive region 212 that does not adhere to the hydrogel laminate 20. The adhesive region 211 has a first adhesive region 211A and a second adhesive region 211B via the non-adhesive region 212.

[0103] The first adhesive region 211A and the second adhesive region 211B have the same configuration as the adhesive region 11 and the non-adhesive region 12 described above. A sensor unit 240 is provided at the interface between the solid substrate 200 and the hydrogel laminate 20, that is, on one surface 200a of the solid substrate 200.

[0104] The sensor unit 240 is provided on the adhesive region 211 and the non-adhesive region 212. The sensor unit 240 has a plurality of unit sensor unit rows 242, each of which is made up of two or more unit sensor units 241 lined up adjacent to each other in the width direction of the adhesive region 211 and the non-adhesive region 212. Here, an example is shown in which the sensor unit 240 has a first unit sensor unit row 242A, a second unit sensor unit row 242B, and a third unit sensor unit row 242C, each of which is made up of unit sensor units 241. The first unit sensor unit row 242A, the second unit sensor unit row 242B, and the third unit sensor unit row 242C are lined up at a distance from each other in the length direction of the adhesive region 211 and the non-adhesive region 212. [Example]

[0105] The present invention will be explained in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0106] [Example] <Detection of analyte solution diffusion using a hydrogel flow channel device with a sensor that uses a thin gold film as the sensing surface of the sensor> The solid substrate with a sensor (hereinafter referred to as "glass substrate with gold thin film"), which uses a gold thin film as the detection surface of the sensor part, was prepared by sputtering gold onto the center of one side of a glass slide.

[0107] The glass substrate with the gold thin film was cleaned by oxygen plasma treatment. Thereafter, a sodium alginate solution was dropped onto the surface of the glass substrate on which the gold thin film had been formed, and then the surface of the glass substrate on which the gold thin film had been formed was spin-coated with the sodium alginate solution to obtain a spin-coated glass substrate with a gold thin film.

[0108] The glass substrate with the spin-coated gold thin film was immersed in an aqueous calcium chloride solution, and then the glass substrate with the spin-coated gold thin film was washed with ultrapure water and dried to form a calcium alginate thin film as a sacrificial layer, thereby obtaining a glass substrate with a calcium alginate thin film.

[0109] A PMMA thin film and a positive photoresist thin film were laminated in this order by spin coating on one side of the calcium alginate thin film-coated glass substrate. Next, a thin film of positive photoresist was shaped into a flow channel by UV exposure through a photomask and development.

[0110] Next, the PMMA thin film and calcium alginate thin film were removed by oxygen plasma etching, and the solid substrate was immersed in a toluene solution containing 25 mmol / L TMSPMA and 25 mmol / L Bis-thiol to form an adhesive area on one side of the solid substrate.

[0111] Next, the solid substrate was immersed in acetone to remove the PMMA thin film and the positive photoresist thin film, thereby obtaining a solid substrate having one side composed of the sacrificial layer and the adhesive region.

[0112] Next, 80 μm thick spacers were placed on both end faces of the solid substrate with the sacrificial layer obtained by the above method, and an aqueous solution consisting of acrylamide, methylenebisacrylamide, and LAP was dropped onto the solid substrate with the sacrificial layer as a precursor solution for the swellable film-like gel.

[0113] One side of the solid substrate with the sacrificial layer was covered with a cover glass, and the precursor solution was gelled by irradiating it with light having a wavelength of 365 nm, thereby forming a swellable gel thin film layer. After the swellable gel thin film layer was formed, the cover glass covering one side of the solid substrate with the sacrificial layer was removed, and unreacted gel precursor molecules were removed in pure water.

[0114] Next, the solid substrate with the sacrificial layer was immersed in a 10 mmol / L EDTA aqueous solution, which served as a dissolution stimulus. This dissolved the sacrificial layer, which consisted of a calcium alginate thin film. The swellable gel thin film layer on top of the sacrificial layer then peeled off from the solid substrate and swelled, forming a hydrogel flow path.

[0115] Next, liquid supply tubes were inserted into both ends of the hydrogel channel, and the liquid supply tubes were bonded and fixed between the swellable gel thin film and the solid substrate using an adhesive.

[0116] A rhodamine solution was delivered from the delivery tube into the hydrogel channel, and the time-dependent change in SPR signal intensity near the inner wall of the hydrogel channel was measured.

[0117] FIG. 20 shows the results of measuring the diffusion of a rhodamine solution in a hydrogel flow channel device equipped with an SPR sensor using an SPR sensor. From the results shown in FIG. 20, it can be confirmed that the intensity of the fluorescence derived from rhodamine inside the hydrogel laminate increases as time passes from t1 to t6.

[0118] Although an embodiment of the present invention has been described in detail above with reference to the drawings, the specific configuration is not limited to this embodiment, and includes designs within the scope of the gist of the present invention. [Industrial Applicability]

[0119] The sensor-equipped hydrogel flow channel device of the present invention is useful as a cell culture device, microreactor, or sensing device that takes advantage of its diffusive flow channel shape, and is widely applicable to fields such as pharmacology, tissue engineering, and chemical engineering. [Explanation of symbols]

[0120] 1 Hydrogel flow channel device with sensor, 10 Solid substrate, 11 Adhesion region, 12 Non-adhesion region, 20 Hydrogel laminate, 21 Swellable gel thin film layer, 22 Non-swellable gel layer, 30 Hydrogel flow channel, 40 Sensor part, 41 Unit sensor part, 42 Unit sensor part array, 43 Detection surface, 44, 45 Probe, 50 Liquid delivery tube

Claims

1. A solid substrate and a hydrogel laminate positioned on the solid substrate, the solid substrate has, at an interface with the hydrogel laminate, an adhesive region that is adhered to the hydrogel laminate and a non-adhesive region that is not adhered to the hydrogel laminate; the hydrogel laminate comprises a swellable gel thin film layer on the solid substrate, and a non-swellable gel layer laminated on the swellable gel thin film layer; a hydrogel flow path formed between the solid substrate and the hydrogel laminate, in which the swellable gel thin film layers are spaced apart in the non-adhesive region; a sensor unit is provided at the interface between the solid substrate and the hydrogel laminate; the sensor unit has a detection surface that detects an analyte near the sensor unit and a probe that specifically binds to the analyte; The hydrogel flow channel type device with a sensor, wherein the detection surface is made of a gold thin film and the probe is made of an antibody.

2. The hydrogel flow channel device with a sensor according to claim 1 , further comprising a liquid supply tube connected to the hydrogel flow channel.

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