WO2018105608A1 - 粒子捕捉デバイス - Google Patents
粒子捕捉デバイス Download PDFInfo
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- WO2018105608A1 WO2018105608A1 PCT/JP2017/043641 JP2017043641W WO2018105608A1 WO 2018105608 A1 WO2018105608 A1 WO 2018105608A1 JP 2017043641 W JP2017043641 W JP 2017043641W WO 2018105608 A1 WO2018105608 A1 WO 2018105608A1
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- WIPO (PCT)
- Prior art keywords
- substrate
- particle
- dispersion medium
- opening area
- particles
- Prior art date
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
- B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N37/00—Details not covered by any other group of this subclass
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L2200/0652—Sorting or classification of particles or molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L2200/0668—Trapping microscopic beads
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
- B01L2300/0829—Multi-well plates; Microtitration plates
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0893—Geometry, shape and general structure having a very large number of wells, microfabricated wells
Definitions
- the present invention relates to a particle capture device.
- This application claims priority on December 7, 2016 based on Japanese Patent Application No. 2016-237237 for which it applied to Japan, and uses the content here.
- Patent Document 1 As a method for comprehensively capturing cells, for example, in Patent Document 1, for the purpose of separating specific cells having different sizes, a substrate having openings having different sizes on an upper surface and a lower surface is used. A substrate and method are described that allow cells smaller than the portion to pass and retain the desired cells in the opening.
- Patent Document 2 has a plurality of openings for isolating and accommodating a single cell as a substrate for capturing and aligning cells on a plane, and cells cannot pass through the bottom of the opening.
- a cell capture substrate having a plurality of sized through-holes is described.
- Patent Document 3 discloses a microwell array having a well of a size that can contain only one cell, and culturing the cell in the microwell array, and detecting a substance produced from the cell stored in the well. Screening methods are described.
- JP-B-2-34597 Japanese Patent No. 2662215 Japanese Patent No. 4148367
- an object of the present invention is to provide a technique for capturing particles uniformly.
- the present invention includes a first substrate and a second substrate disposed to face one side of the first substrate in parallel, and the first substrate includes the first substrate.
- a plurality of recesses that open to the other side of one substrate and have a size capable of capturing one particle, the recesses communicating the one side and the other side, and the dispersion medium for the particles Has a communication hole having a movable size, and between the first substrate and the second substrate, the communication hole of the first substrate is used as the inlet of the dispersion medium,
- a flow path is formed in which an end portion on one side of the first substrate is an outlet of the dispersion medium, and a total opening area of the communication holes is 1 mm 2 or more and less than 10 mm 2 , and The cross-sectional area of the flow path is 0.8 times or more of the total opening area of the communication holes, or the communication holes are opened. Total is 10 mm 2 or more 1000 mm 2 or less and the cross-sectional area of the flow passage in the outlet port at least 0.1 times the
- the present invention includes a first substrate and a second substrate disposed to face one side of the first substrate in parallel, and the first substrate includes the first substrate.
- a plurality of recesses that open to the other side of one substrate and have a size capable of capturing one particle, the recesses communicating the one side and the other side, and the dispersion medium for the particles Has a communication hole having a movable size, and between the first substrate and the second substrate, the communication hole of the first substrate is used as the inlet of the dispersion medium, A flow path having an end on one side of the first substrate as an outlet of the dispersion medium is formed, a total opening area of the communication holes is 1 mm 2 or more, and the first substrate and the It is a particle
- the present invention includes a first substrate and a second substrate disposed to face one side of the first substrate in parallel, and the first substrate includes the first substrate.
- a plurality of recesses that open to the other side of one substrate and have a size capable of capturing one particle, the recesses communicating the one side and the other side, and the dispersion medium for the particles Has a communication hole having a movable size, and between the first substrate and the second substrate, the communication hole of the first substrate is used as the inlet of the dispersion medium,
- a flow path is formed in which an end portion on one side of the first substrate is an outlet of the dispersion medium, and a cross-sectional area of the flow path at the outlet is 0 of a total opening area of the communication holes.
- a particle trapping device that is 8 times or more.
- the present invention includes a first substrate and a second substrate disposed to face one side of the first substrate in parallel, and the first substrate includes the first substrate.
- a plurality of recesses that open to the other side of one substrate and have a size capable of capturing one particle, the recesses communicating the one side and the other side, and the dispersion medium for the particles Has a communication hole having a movable size, and between the first substrate and the second substrate, the communication hole of the first substrate is used as the inlet of the dispersion medium,
- a flow path is formed in which an end portion on one side of the first substrate is an outlet of the dispersion medium, and a total opening area of the communication holes is 1 mm 2 or more and less than 10 mm 2 , and The cross-sectional area of the flow path is 0.8 times or more of the total opening area of the communication holes, or the communication holes are opened.
- Total is 10 mm 2 or more 1000 mm 2 or less and the cross-sectional area of the flow passage in the outlet port at least 0.1 times the total opening area of the communication hole of the area, to provide a particle capture device.
- particles can be captured uniformly.
- the particle capturing device includes a first substrate and a second substrate disposed to face one side of the first substrate in parallel, and the first substrate includes the first substrate.
- a plurality of recesses that open to the other side of one substrate and have a size capable of capturing one particle, the recesses communicating the one side and the other side, and the dispersion medium for the particles Has a communication hole having a movable size, and between the first substrate and the second substrate, the communication hole of the first substrate is used as the inlet of the dispersion medium, A flow path having an end on one side of the first substrate as an outlet of the dispersion medium is formed, a total opening area of the communication holes is 1 mm 2 or more, and the first substrate and the The distance between the second substrate and the second substrate may be 100 ⁇ m or more. As will be described later in the Examples, particles can be uniformly captured by such a particle capturing device.
- FIG. 1 (a) and (b), FIG. 2, FIG. 3 (a) to (d), FIG. 4 (a) and (b), and FIG. 5 (a) and (b) are the particles of this embodiment, respectively.
- FIG. 1A is a front sectional view
- FIG. 1B is a top view
- 2 and 3A to 3D are perspective views
- 4A is a front sectional view
- FIG. 4B is a top view
- 5A is a front cross-sectional view
- FIG. 5B is a top view.
- the particle trapping device 100 of the present embodiment includes a first substrate 10 and a second substrate 20 disposed so as to face the one side 11 of the first substrate 10 in parallel.
- the first substrate 10 has a plurality of recesses 13 that are open on the other side 12 of the first substrate 10 and have a size capable of capturing one particle.
- the recessed part 13 has the communicating hole 14 which connects the one side 11 and the other side 12, and has a magnitude
- the communication holes 14 of the first substrate 10 are used as an inlet for the dispersion medium, and the end portions 11 a and 11 b on one side 11 of the first substrate 10. Is formed as a flow path 30.
- the total cross-sectional area of the flow path 30 at the outlets 11a and 11b is the opening of the communication hole 14 when the total opening area of the communication holes 14 is 1 mm 2 or more and less than 10 mm 2 , for example, 2 to 8 mm 2. 0.8 times or more of the total area.
- the total cross-sectional area of the flow path 30 at the outlets 11a and 11b is such that the total opening area of the communication holes 14 is 10 mm 2 or more and 1000 mm 2 or less, such as 10 to 500 mm 2 , such as 10 to 300 mm 2 , such as 10 to 100 mm. 2 , for example, 10 to 50 mm 2 , is 0.1 times or more of the total opening area of the communication holes 14.
- the total opening area of the communication holes 14 is 1 mm 2 or more, such as 1 to 1000 mm 2 , such as 1 to 500 mm 2 , such as 1 to 300 mm 2 , such as 1 to 100 mm 2 , 1 to 50 mm 2 , and the distance between the first substrate and the second substrate may be 100 ⁇ m or more.
- the area of the outlet is the sum of the cross-sectional areas of the flow path 30 at the end portion (11 a and 11 b) on the one side 11 of the first substrate 10.
- the particle trapping device is configured such that when the total opening area of the communication holes 14 is 1 mm 2 or more and less than 10 mm 2 , the area of the outlet is the total opening area of the communication holes 14. By being 0.8 times or more, the particles can be captured uniformly. Further, in the particle capturing device of the present embodiment, when the total opening area of the communication holes 14 is 10 mm 2 or more and 1000 mm 2 or less, the area of the outlet is 0.1 times or more of the total opening area of the communication holes 14. As a result, the particles can be captured uniformly. Further, in the particle capturing device of the present embodiment, the total opening area of the communication holes 14 is 1 mm 2 or more, and the distance between the first substrate and the second substrate is 100 ⁇ m or more. It can be captured uniformly.
- the particle capturing rate is small at the center of the particle capturing device, and the end of the particle capturing device (portion close to the outlet) Then, there are cases where the trapping rate of particles increases. More specifically, the photograph shown in FIG. 7 is such an example.
- the particle trapping rate refers to the ratio of the number of recesses capturing particles to the total number of recesses included in the unit region.
- the unit region is not particularly limited, and may be, for example, one field of view when observed with a microscope.
- the particle trapping device of this embodiment there is little variation between the particle trapping rate at the center of the particle trapping device and the particle trapping rate at the end of the particle trapping device. Therefore, according to the particle trapping device of the present embodiment, particles can be trapped uniformly throughout the entire particle trapping device.
- the particle capture rate is uniform is synonymous with “small variation in the capture rate”
- the ratio of the particle capture rate between arbitrary regions of the particle capture device is, for example, It means 0.7 or more, preferably 0.8 or more, more preferably 0.9 or more.
- the particle is not particularly limited, and examples thereof include a cell, a cell mass, a resin particle, a metal particle, a glass particle, and a ceramic particle.
- the diameter of the particles is not particularly limited, and may be, for example, about 1 to 500 ⁇ m, for example, about 1 to 200 ⁇ m, for example, about 1 to 100 ⁇ m, for example, about 1 to 50 ⁇ m. Also good. In this specification, the diameter of a particle refers to the diameter of a circle having the same area as the projected area of the particle.
- the dispersion medium In capturing the particles, the particles are supplied from the other side 12 of the first substrate 10 while being suspended in the dispersion medium.
- the dispersion medium is not particularly limited, and examples thereof include water, a buffer solution, an isotonic solution, a culture medium, and the like.
- the first substrate 10 may be formed of a layer 10a in which the recess 13 is patterned and a layer 10b in which the communication hole 14 is patterned.
- the substrate 10 may have a structure in which a plurality of recesses 13 are arranged vertically and horizontally at the same interval.
- B represents one particle.
- the shape of the recess 13 is not particularly limited as long as it can capture one particle.
- the shape of the recess 13 may be a cylindrical shape, may be a polyhedron (for example, a rectangular parallelepiped, a hexagonal column, an octagonal column, etc.) formed of a plurality of surfaces, may be an inverted truncated cone, and may be reversed.
- truncated pyramid an inverted triangular truncated pyramid, an inverted quadrangular truncated pyramid, an inverted pentagonal truncated pyramid, an inverted hexagonal truncated pyramid, an inverted polygonal truncated pyramid with a heptagon or more
- truncated pyramid an inverted triangular truncated pyramid, an inverted quadrangular truncated pyramid, an inverted pentagonal truncated pyramid, an inverted hexagonal truncated pyramid, an inverted polygonal truncated pyramid with a heptagon or more
- the shape of the recess 13 may be, for example, a part of a cylinder and the rest of an inverted truncated cone.
- the shape of the concave portion 13 is a cylindrical shape or a rectangular parallelepiped
- the bottom portion of the concave portion 13 is usually flat, but may be a curved surface (convex surface or concave surface).
- the dimensions of the recess 13 can be appropriately determined in consideration of a suitable ratio between the diameter of the particles to be captured in the recess 13 and the dimension of the recess 13.
- the recess 13 is preferably patterned so that its form, density, and the like are controlled.
- the shape and size of the recess 13 are appropriately determined so that one particle is captured in one recess 13 in consideration of the type of particles to be captured in the recess 13 (particle shape, size, etc.). Can be determined.
- the diameter of the largest circle inscribed in the planar shape of the recess 13 is 0.5 to 0.5 of the diameter of the particle to be trapped in the recess 13.
- a range of 2 times is preferable, a range of 0.8 to 1.9 times is more preferable, and a range of 0.8 to 1.8 times is more preferable.
- the depth of the recess 13 is preferably in the range of 0.5 to 4 times the diameter of the particles to be trapped in the recess 13 and more preferably in the range of 0.8 to 1.9 times.
- the range of 0.8 to 1.8 times is more preferable.
- the thickness of the first substrate 10 is preferably as follows.
- the thickness of the first substrate 10 is preferably 1 to 100 ⁇ m, and more preferably 10 to 50 ⁇ m.
- the number of the recesses 13 included in the first substrate 10 is not particularly limited, but is preferably in the range of, for example, 2,000 to 1,000,000 per 1 cm 2 .
- the opening ratio of the recessed part 13 may be less than 100% from the technical problem on manufacture.
- the opening ratio of the recess 13 is preferably in the range of 1 to 90%, for example.
- the size of the recess 13 is preferably 1 to 100 ⁇ m in diameter, more preferably 2 to 50 ⁇ m, and further preferably 3 to 25 ⁇ m.
- the depth of the recess 13 is preferably 1 to 100 ⁇ m, more preferably 2 to 70 ⁇ m, still more preferably 3 to 50 ⁇ m, and particularly preferably 4 to 30 ⁇ m.
- the depth of the recess 13 is 1 ⁇ m or more, it is easy to capture particles, which is preferable from the viewpoint of practical use.
- the depth of the recessed part 13 is 100 micrometers or less, it is preferable from a viewpoint with a low possibility that several particle
- the dimension of the communication hole 14 can be appropriately determined in consideration of the diameter of the particle to be captured in the recess 13, the dimension of the recess 13, the characteristics of the dispersion medium of the particles to be moved through the communication hole 14, and the like.
- the communication holes 14 are preferably patterned so that the form, the diameter of the pores, the density thereof, and the like are controlled. Controlling the communication holes is preferable because it is easy to ensure the uniformity of the amount of the particle dispersion medium permeated.
- the communication holes 14 are not limited to those produced by patterning, and for example, those formed using a porous material such as a porous film can also be used.
- the number, position, shape, size, and the like of the communication holes 14 can be captured (stored inside the recess 13) without passing through the particles, and the dispersion medium can move.
- the dispersion medium can move.
- a plurality of cylindrical communication holes 14 having a diameter smaller than the diameter of the recess 13 may be provided at the bottom of the recess 13.
- a communication hole having a shape as shown in 14a to 14d of FIGS. 3 (a) to 3 (d) is provided at the bottom of the recess 13. Also good.
- the diameter of the communication hole 14 is preferably 10 nm to 20 ⁇ m, and preferably 50 nm to 15 ⁇ m. More preferred is 100 nm to 10 ⁇ m.
- the width is preferably 10 nm to 20 ⁇ m, more preferably 50 nm to 15 ⁇ m, still more preferably 100 nm to 10 ⁇ m.
- the communication holes 14 are in a lattice shape, one side is preferably 10 nm to 20 ⁇ m, more preferably 50 nm to 15 ⁇ m, still more preferably 100 nm to 10 ⁇ m.
- the cross-sectional area of the surface parallel to the first substrate of the communication port 14 is constant throughout the communication port 14.
- the cross-sectional area of the plane parallel to the first substrate at an arbitrary position of the communication port 14 may be the opening area of the communication port 14.
- the opening area of the communication port 14 is the smallest cross-sectional area of the cross-sectional area of the surface parallel to the first substrate.
- the opening area of the communication port 14 may be used.
- the total opening area of the communication ports 14 is the total area of the opening areas of all the communication ports 14 included in the particle supplement device of the present embodiment.
- the particle trapping device of the present embodiment includes a second substrate 20 arranged so as to face the one side 11 of the first substrate 10 in parallel. Moreover, between the 1st board
- the flow path 30 is formed.
- a pillar 22 that supports the first substrate 10 may exist between the first substrate 10 and the second substrate 20.
- the number, position, shape, size, etc. of the pillars 22 are not particularly limited as long as the first substrate 10 is supported and the object of the present invention can be achieved.
- the area of the outlet may be larger than the total opening area of the communication holes 14.
- the area of the outlet is the breakage of the flow path 30 at the end portion (11 a and 11 b) on the one side 11 of the first substrate 10. The total area.
- the opening area of the communication hole 14 can be obtained based on the porosity of the porous material. More specifically, for example, the product of the total opening area of the recess 13 and the porosity of the porous material forming the communication hole 14 may be regarded as the total opening area of the communication hole 14.
- the area of the outlet may be 1.2 times or more of the total opening area of the communication holes 14, may be 1.5 times or more, and may be 2 times or more. It may be 2.5 times or more, 3 times or more, 4 times or more, or 5 times or more.
- the distance between the first substrate 10 and the second substrate 20 may be, for example, 100 ⁇ m or more. It may be 150 ⁇ m or more, for example, 200 ⁇ m or more, for example, 250 ⁇ m or more, for example, 300 ⁇ m or more, for example, 350 ⁇ m or more.
- the upper limit of the distance between the first substrate 10 and the second substrate 20 is not limited as the performance of the particle trapping device, but considers practicality (the amount of dispersion medium used, the size of the microscope to be observed, etc.) Then, 5 mm or less is preferable.
- the material of the particle capturing device of the present embodiment is not particularly limited, but is preferably a transparent material from the viewpoint of facilitating particle observation. Furthermore, when observing the captured particles using fluorescence observation as an index, it is preferable to use a material with less autofluorescence.
- Specific materials of the first substrate 10 and the second substrate 20 include, for example, glass, polyethylene terephthalate (PET), polymethyl methacrylate (PMMA), polycarbonate (PC), cycloolefin polymer (COP), epoxy, and the like.
- PET polyethylene terephthalate
- PMMA polymethyl methacrylate
- PC polycarbonate
- COP cycloolefin polymer
- epoxy epoxy
- the material of the particle capturing device of this embodiment has no cytotoxicity and low cell adhesion.
- the material of the particle trapping device of the present embodiment is a curing that is easy to perform microfabrication from the viewpoint of forming a recess 13 having a size capable of capturing one particle and a communication hole 14 having a size capable of moving the dispersion medium.
- the polymer is preferably polymerized using a photosensitive resin composition (hereinafter sometimes referred to as “photosensitive resin composition”).
- the curable resin composition has the property of being cured by crosslinking by irradiating active energy rays such as ultraviolet rays, and is a negative type photoresist, a negative type dry film resist, and a fine resin molding having a fine structure. What is used for etc. is preferable.
- a cured product obtained by curing a curable resin composition into a desired shape by photolithography may be referred to as a resin pattern.
- the curable resin composition When the curable resin composition is used for applications such as micro resin molding, first, the curable resin composition is applied to the surface of the substrate on which the resin pattern is formed, and the solvent component contained in the curable resin composition is applied. The resin film is produced by volatilization. Next, a photomask having a shape of a pattern to be formed is placed on the surface of the resin film and irradiated with active energy rays such as ultraviolet rays. Thereafter, a resin pattern is formed on the surface of the base material through a development step and, if necessary, a post-bake step. This resin pattern can be used for the particle capturing device of the present embodiment.
- Such a curable resin composition includes, for example, an epoxy functional novolak resin, a cationic photopolymerization initiator such as a triarylsulfonium salt, and a diluent capable of reacting with an epoxy functional group, and is completely cured.
- a photo-curable composition that is a resin that is difficult to peel off; a thick film is formed containing polyfunctional bisphenol A formaldehyde-novolak resin, acid generator triphenylsulfonium hexafluoroantimonate, and solvent PGMEA
- a resin composition generally used for fine resin molding, such as a photocurable composition that becomes a possible resin, can be employed.
- a photosensitive (curable) resin composition is prepared by combining an epoxy resin and a specific acid generator, and a resin pattern is formed using this curable resin composition, it is highly sensitive and heat cured. A resin pattern with a small volumetric shrinkage and a high aspect ratio can be formed.
- curable (photosensitive) resin composition examples include a photosensitive resin composition containing (a) a polyfunctional epoxy resin and (b) a cationic polymerization initiator.
- the polyfunctional epoxy resin used in this embodiment is a resin having two or more epoxy groups in one molecule, and a sufficient number of epoxy to cure the resin film formed of the curable resin composition. Any epoxy resin may be used as long as it is an epoxy resin containing a group in one molecule.
- a polyfunctional epoxy resin a phenol novolak type epoxy resin, an ortho cresol novolak type epoxy resin, a triphenyl type novolak type epoxy resin, and a bisphenol A novolak type epoxy resin are preferable.
- the functionality that is the number of epoxy groups contained in one molecule of the polyfunctional epoxy resin is preferably 2 or more, and more preferably 3 to 12.
- the functionality of the polyfunctional epoxy resin is 3 or more, a resin pattern having a high aspect ratio and resolution can be formed, and preferably, the functionality of the polyfunctional epoxy resin is 12 or less. Resin synthesis is easy to control, and it is preferable because the internal stress of the resin pattern can be prevented from becoming excessively large.
- the mass average molecular weight of the polyfunctional epoxy resin is preferably 300 to 5000, and more preferably 500 to 4000.
- the mass average molecular weight of the polyfunctional epoxy resin is preferably 300 or more, which is preferable in that the curable resin composition can be prevented from flowing heat before being cured by irradiation with active energy rays, and the mass of the polyfunctional epoxy resin.
- An average molecular weight of 5,000 or less is preferable in that an appropriate dissolution rate during patterning development can be obtained.
- the content of the polyfunctional epoxy resin in the photosensitive resin composition is preferably 10 to 99.9% by mass and more preferably 30 to 99.9% by mass in the total solid content. As a result, a photosensitive resin film having high sensitivity and appropriate hardness can be obtained when coating on the substrate.
- a cationic polymerization initiator is a compound that generates cations upon irradiation with active energy rays such as ultraviolet rays, far ultraviolet rays, excimer laser light such as KrF and ArF, X-rays and electron beams, and the cations can serve as polymerization initiators. is there.
- Examples of such cationic polymerization initiators include 4- (2-chloro-4-benzoylphenylthio) phenyldiphenylsulfonium hexafluoroantimonate, 4- (2-chloro-4-benzoylphenylthio) phenylbis (4 -Fluorophenyl) sulfonium hexafluoroantimonate, 4- (2-chloro-4-benzoylphenylthio) phenylbis (4-chlorophenyl) sulfonium hexafluoroantimonate, 4- (2-chloro-4-benzoylphenylthio) phenyl Bis (4-methylphenyl) sulfonium hexafluoroantimonate, 4- (2-chloro-4-benzoylphenylthio) phenylbis (4- ( ⁇ -hydroxyethoxy) phenyl) sulfonium hexafluoroanti
- the content of the cationic polymerization initiator in the curable resin composition is preferably 0.1 to 10% by mass, and more preferably 0.5 to 5% by mass. If the content of the cationic polymerization initiator in the curable resin composition is 0.1% by mass or more, it is preferable because the curing time of the curable resin composition by active energy ray exposure can be made appropriate. Moreover, if the content of the cationic polymerization initiator in the curable resin composition is 10% by mass or less, the developability after exposure with active energy rays can be improved, which is preferable. In addition, the said content is a thing when the curable resin composition does not contain the solvent component mentioned later.
- the content of the cationic polymerization initiator after removing the mass of the solvent component may be within the above range.
- the details of the curable resin composition can be carried out based on methods known to those skilled in the art described in JP 2008-180877 A, JP 2011-111588 A, and the like. To be understood.
- the shape of the first substrate 10, the shape of the second substrate 20, and the arrangement of the first substrate 10 and the second substrate 20 are not limited to those in FIG. .
- the first substrate 10 and the second substrate 20 are both rectangular, but the first substrate 10 and the second substrate 20 may be circular, for example, triangular. Polygons such as pentagons, hexagons, heptagons, and octagons may be used.
- the first substrate 10 is arranged in the center of the second substrate 20 and there are two outlets 11a and 11b.
- the second substrate 20 and one end may be arranged at the same position, and only one of the outlets 11a or 11b may exist.
- 4A and 4B are schematic views showing an example of the particle trapping device of the present embodiment.
- 4A is a front sectional view
- FIG. 4B is a top view.
- the particle capturing device 400 includes a first substrate 10 and a second substrate 20 disposed so as to face the one side 11 of the first substrate 10 in parallel. Further, the first substrate 10 has a plurality of recesses 13 that are open on the other side 12 of the first substrate 10 and have a size capable of capturing one particle. Moreover, the recessed part 13 has the communicating hole 14 which connects the one side 11 and the other side 12, and has a magnitude
- the communication hole 14 of the first substrate 10 is used as an inlet for the dispersion medium, and the end portion 11 a on one side 11 of the first substrate 10 is the above-mentioned
- a flow path 30 is formed as an outlet for the dispersion medium.
- the area of the outflow port 11a is 0.8 times or more of the total opening area of the recess 13.
- the relative position between the first substrate 10 and the second substrate 20 is determined by the holding member 40.
- the particle capturing device 400 may be configured to hold the substrate 10 on the substrate 20 by adding a pillar or the like to the lower portion of the substrate 10 instead of the holding member 40.
- the planar shape of the first substrate 10 is circular.
- the planar shape of the second substrate 20 is also circular. For this reason, the particle trapping device shown in FIG. 4 is shaped like a petri dish with two layers.
- the end 11 a on one side 11 of the first substrate 10 is the circumference of the circular first substrate 10.
- the area of the outlet is the cross-sectional area of the flow path 30 on the circumference 11 a of the first substrate 10.
- FIGS. 5A and 5B are schematic views illustrating an example of the particle trapping device of the present embodiment.
- FIG. 5A is a front sectional view
- FIG. 5B is a top view.
- the particle trapping device 500 includes a first substrate 10 and a second substrate 20 arranged to face one side 11 of the first substrate 10 in parallel. Further, the first substrate 10 has a plurality of recesses 13 that are open on the other side 12 of the first substrate 10 and have a size capable of capturing one particle. Moreover, the recessed part 13 has the communicating hole 14 which connects the one side 11 and the other side 12, and has a magnitude
- the particle trapping device 500 may be configured to hold the substrate 10 on the substrate 20 by adding a pillar or the like to the lower portion of the substrate 10 instead of the holding member 40.
- the planar shape of the first substrate 10 is a rectangle.
- the planar shape of the second substrate 20 is a circle.
- the end portions 11 a, 11 b, 11 c, and 11 d on one side 11 of the first substrate 10 are one side of the outer periphery of the rectangular first substrate 10.
- the area of the outlet is the sum of the cross-sectional areas of the flow paths 30 in the outer sides 11 a, 11 b, 11 c, and 11 d of the first substrate 10.
- a plurality of the particle trapping devices described above may be connected.
- a plurality of the above-described particle capturing devices 400 or 500 are connected to each other to form a 6-well plate type, a 12-well plate type, a 24-well plate type, a 48-well plate type, a 96-well plate type, a 384-well plate type, and a 1536-well plate.
- You may form in shapes, such as a type
- the size of the particle capturing device is a size that conforms to the SBS standard, a slide glass size, or a petri dish size that is widely used for cell culture and the like from a practical viewpoint. It is preferable to produce it.
- the present invention provides a method for manufacturing the particle capture device described above.
- a dissolvable base film is formed on a first support, and the first curable resin film is applied by applying a first curable resin composition on the base film.
- a particle trapping device is a bonded product.
- the second substrate may have a pillar.
- the third curable resin composition is applied on the second substrate to form the third curable resin film, and the third curable resin film is formed.
- a step a may be further included in which a pillar is patterned on the curable resin film to obtain a second substrate on which the pillar is patterned.
- a dissolvable base film 32 is formed on the first support 31, and the first curable resin composition is formed on the base film 32.
- the first curable resin film 10B is applied to form a layer 10b in which the first curable resin film 10B is exposed and then developed to pattern the communication holes 14 as shown in FIG. 6C. Form.
- the patterning method of the communication hole 14 is not limited to exposure and development, and an imprint method or a method using a guided self assembly (DSA) technique may be employed. Moreover, the hardening method of the 1st curable resin film 10B may not be exposure, and a well-known method is employ
- DSA guided self assembly
- Examples of the first support include, for example, a substrate for electronic components, a substrate on which a predetermined wiring pattern is formed, and the like. More specifically, a silicon substrate, a metal substrate such as copper, chromium, iron, and aluminum, a glass substrate, and the like can be given. As a material for the wiring pattern, for example, copper, aluminum, nickel, gold or the like can be used.
- Examples of the first curable resin composition include the curable (photosensitive) resin compositions described above.
- the base film 32 includes polyvinyl alcohol resin, dextrin, gelatin, glue, casein, shellac, gum arabic, starch, protein, polyacrylic acid amide, sodium polyacrylate, polyvinyl methyl ether, styrene elastomer, methyl vinyl ether and anhydrous maleic acid.
- a copolymer with an acid, a copolymer of vinyl acetate and itaconic acid, polyvinyl pyrrolidone, acetyl cellulose, hydroxyethyl cellulose, sodium alginate and the like can be used. These materials may be a combination of a plurality of materials soluble in the same kind of liquid.
- the material of the base film may include a rubber component such as mannan, xanthan gum, or guar gum.
- the second curable resin film 10A is formed on the layer 10b by applying the second curable resin composition, and the second curable resin composition is formed.
- the resin film 10A is exposed and then developed to obtain the first substrate 10 in which the recesses 13 are patterned on the layer 10b.
- Examples of the second curable resin composition include the curable (photosensitive) resin compositions described above.
- the patterning method of the concave portion 13 is not limited to exposure / development, and an imprint method or a method using a guided self assembly (DSA) technique can also be employed.
- the hardening method of the 2nd curable resin composition may not be exposure but a well-known method is employ
- the base film 32 is dissolved by immersing the entire substrate in a release agent (for example, 1-methyl-4-isopropylcyclohexane (p-Menthane)), and the first substrate 31 to the first substrate. 10 is peeled off.
- a release agent for example, 1-methyl-4-isopropylcyclohexane (p-Menthane)
- the first substrate 10 shown in FIG. 6 (f) obtained in the above step and the second substrate 20 shown in FIG. 6 (g) are bonded.
- the layer 10 b is bonded so as to face the second substrate 20.
- the curable resin composition may be used as an adhesive.
- the second substrate 20 may have pillars 22.
- the third curable resin composition is applied on the second support 20 to form the third curable resin film 22A.
- a pillar pattern 22 as shown in FIG. 6G is formed.
- the formation of the pillar pattern 22 is optional, and this step may not exist.
- the hardening method of the 3rd curable resin composition may not be exposure but a well-known method is employ
- the support 20 for example, a substrate for electronic components can be used, but a transparent substrate is preferable from the viewpoint of easy observation of captured particles, and specifically, a glass substrate is preferably used.
- the third curable resin composition include the curable (photosensitive) resin compositions described above.
- the present invention provides a particle trapping method comprising supplying particles to the inlet of the particle trapping device described above and causing the dispersion medium to flow out from the outlet.
- the capturing method of the present embodiment can be said to be a method for capturing particles uniformly, a method for producing uniformly captured particles, or the like.
- the particles supplied from the inlet of the particle capturing device described above are captured in the recess 13 provided in the first substrate 10. Further, the particle dispersion medium moves through the communication hole 14, passes through the flow path 30, and is discharged from the outlet.
- particles can be captured uniformly by using the particle capturing device described above.
- Example 1 Manufacture of the first substrate
- a spin coater (1500 rpm, 20 seconds)
- prebaked 90 ° C. for 1 minute and 120 ° C. for 3 minutes by a hot plate to form a base film.
- a photosensitive resin composition (see Japanese Patent Application Laid-Open Nos. 2008-180877 and 2011-111588) is applied onto the base film with a spin coater (3000 rpm, 20 seconds), and is heated at 90 ° C. for 3 minutes with a hot plate. Pre-baked. Thereafter, pattern exposure (GHI line, 150 mJ) was performed using a mirror projection mask aligner (model “MPA-600FA”, manufactured by Canon Inc.), and heating was performed after exposure at 90 ° C. for 5 minutes using a hot plate. Thereafter, development processing for 30 seconds was performed by an immersion method using propylene glycol monomethyl ether acetate (PGMEA). Next, the developed resin pattern was post-baked with the substrate at 120 ° C. for 1 minute using an oven to obtain a cylindrical communication hole resin pattern.
- PGMEA propylene glycol monomethyl ether acetate
- the photosensitive resin composition was applied onto the communication hole resin pattern obtained above with a spin coater (1000 rpm, 20 seconds), and prebaked at 90 ° C. for 5 minutes with a hot plate. Thereafter, pattern exposure (GHI line, 60 mJ) was performed using a mirror projection mask aligner (model “MPA-600FA”, manufactured by Canon Inc.), and heating was performed after exposure at 90 ° C. for 5 minutes using a hot plate. Then, the development process for 2 minutes was performed by the immersion method using PGMEA. Next, the developed resin pattern was post-baked for 1 minute at 120 ° C. using an oven together with the substrate to obtain a concave pattern. The recesses were cylindrical with a diameter of 10 ⁇ m.
- the first substrate in which the concave portion obtained as described above is patterned is immersed in a release agent, and the base film is dissolved, whereby the first substrate in which the concave portion pattern is formed on the communication hole resin pattern from the silicon substrate.
- the substrate was peeled off.
- the said photosensitive resin composition was apply
- the resin pattern may be referred to as a distance between the first substrate and the second substrate (hereinafter referred to as “flow path height”) when the first substrate and the second substrate are bonded in a process described later. .), And a resin pattern having a distance between the first substrate and the second substrate of 120 ⁇ m was produced.
- the flow path height was high (for example, 100 micrometers or more)
- the said coating process by a spin coater was repeatedly performed until it became target height.
- the thickness of the first substrate was 10 ⁇ m, the pitch between the recesses was 75 ⁇ m, and the diameter of the cylindrical communication hole was 2 ⁇ m.
- the diameter of the recess was 10 ⁇ m, and the distance between the first substrate and the second substrate was 120 ⁇ m.
- the size of the particle trapping device was a rectangular shape having a length of 75 mm and a width of 26 mm in the plan view, and the thickness was 15 mm.
- the area of the outlet was 4.8 mm 2 .
- the area of the outlet is the sum of the cross-sectional areas of the flow passages 30 at two locations 11a and 11b shown in FIG. Further, the total opening area of the communication holes was 2.7 mm 2 . Therefore, the area of the outlet was about 1.78 times the total opening area of the communication holes.
- Example 2 to 11 Comparative Examples 1 and 2
- Examples 2 to 8 and Comparative Example were the same as Example 1 except that the device shape, the diameter of the recess, the flow path height, the total area of the recess communication holes, and the outlet area were as shown in Table 1. 1-2 particle capture devices were made.
- the particle trapping device having a rectangular device shape was a rectangular shape having a length of 75 mm and a width of 26 mm in a plan view, and a thickness of 15 mm.
- the particle trapping device having a circular device shape was a circular shape having an outer diameter of 5.3 mm and a thickness of 1.3 mm.
- Table 1 shows the measurement results of the particle trapping devices of Examples 1 to 11 and Comparative Examples 1 and 2.
- “Area ratio” indicates the ratio of the area of the outlet to the total opening area of the communication holes of the particle trapping device.
- “cell number ratio” indicates the ratio of the number of cells in the central portion to the number of cells in the end portion of the particle capturing device in one visual field when observed with a fluorescence microscope. This value is the same as the ratio of the cell capture rate at the center to the cell capture rate at the end of the particle capture device.
- FIG. 7 is a photograph showing, as an example, the result of fluorescence microscope observation of the central part and the end part of the particle capturing device after capturing the Namalwa cells with the particle capturing device of Comparative Example 1. As shown in FIG. 7, in the particle capturing device of Comparative Example 1, the cell capturing rate varied depending on the position of the particle capturing device.
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Abstract
Description
1実施形態において、本発明は、第1の基板と、前記第1の基板の一方側に平行に対向するように配置された第2の基板とを備え、前記第1の基板は、前記第1の基板の他方側に開口し、粒子1個を捕捉可能な大きさを有する凹部を複数有しており、前記凹部は、前記一方側と前記他方側とを連通し、前記粒子の分散媒が移動可能な大きさを有する連通孔を有しており、前記第1の基板と前記第2の基板との間は、前記第1の基板の前記連通孔を前記分散媒の流入口とし、前記第1の基板の一方側の端部を前記分散媒の流出口とする流路を形成しており、前記連通孔の開口面積の合計が1mm2以上10mm2未満であり前記流出口における前記流路の断面積が前記連通孔の開口面積の合計の0.8倍以上であるか、又は、前記連通孔の開口面積の合計が10mm2以上1000mm2以下であり前記流出口における前記流路の断面積が前記連通孔の開口面積の合計の0.1倍以上である、粒子捕捉デバイスを提供する。実施例において後述するように、本実施形態の粒子補足デバイスによれば、粒子を均一に捕捉することができる。
本実施形態の粒子捕捉デバイスにおいて、粒子としては特に制限されず、例えば、細胞、細胞塊、樹脂粒子、金属粒子、ガラス粒子、セラミック粒子等が挙げられる。粒子の直径は特に制限されず、例えば約1~500μmであってもよく、例えば約1~200μmであってもよく、例えば約1~100μmであってもよく、例えば約1~50μmであってもよい。本明細書において、粒子の直径とは、粒子の投影面積と同じ面積の円の直径をいうものとする。
粒子を捕捉するにあたり、粒子は分散媒に懸濁された状態で第1の基板10の他方側12から供給される。分散媒としては、特に制限されず、水、緩衝液、等張液、培地等が挙げられ、目的に応じて適宜用いることができる。
図1(a)に示すように、第1の基板10は、凹部13がパターニングされた層10aと、連通孔14がパターニングされた層10bとから形成されていてもよい。基板10は、例えば図2に示すように、複数個の凹部13が同一間隔で縦横に配置された構造を有していてもよい。
連通孔14の寸法は、凹部13に捕捉しようとする粒子の直径と、凹部13の寸法と、連通孔14を移動させるべき粒子の分散媒の特性等を考慮して適宜決定することができる。連通孔14は、パターニングされ、形態、細孔の径、その密度等が制御されていることが好ましい。連通孔が制御されている場合、粒子の分散媒の透過量の均等性を確保しやすいため好ましい。しかしながら、連通孔14としては、パターニングにより作製されたものに限られず、例えば、多孔質膜等の多孔質材料を使用して形成したものも使用することができる。
例えば、連通口14が円筒状である場合、連通口14の第1の基板と平行な面の断面積は、連通口14の全体にわたって一定である。この場合、連通口14の任意の位置における、第1の基板と平行な面の断面積を連通口14の開口面積とすればよい。
図1(a)に示すように、本実施形態の粒子捕捉デバイスは、第1の基板10の一方側11に平行に対向するように配置された第2の基板20を備えている。また、第1の基板10と第2の基板20との間は、第1の基板10の連通孔14を流入口とし、第1の基板10の一方側11の端部11a及び11bを流出口とする流路30を形成している。
本実施形態の粒子捕捉デバイスの材質は、特に限定されないが、粒子の観察を容易とする観点から、透明性のある材質であることが好ましい。更に、捕捉した粒子を、蛍光観察を指標として観察する場合には、自家蛍光の少ない材質であることが好ましい。
本実施形態で使用される多官能エポキシ樹脂は、1分子中に2個以上のエポキシ基を有する樹脂であり、硬化性樹脂組成物で形成された樹脂膜を硬化させるのに十分な数のエポキシ基を1分子中に含むエポキシ樹脂であれば、どのようなエポキシ樹脂でもよい。このような多官能エポキシ樹脂としては、フェノールノボラック型エポキシ樹脂、オルトクレゾールノボラック型エポキシ樹脂、トリフェニル型ノボラック型エポキシ樹脂、ビスフェノールAノボラック型エポキシ樹脂が好ましい。
次に、カチオン重合開始剤について説明する。カチオン重合開始剤は、紫外線、遠紫外線、KrF、ArF等のエキシマレーザー光、X線、電子線等の活性エネルギー線の照射を受けてカチオンを発生し、そのカチオンが重合開始剤となり得る化合物である。
本実施形態の粒子捕捉デバイスにおいて、第1の基板10の形状、第2の基板20の形状、第1の基板10及び第2の基板20の配置は図1(b)のものに限られない。例えば、図1(b)では、第1の基板10及び第2の基板20はいずれも矩形であるが、第1の基板10、第2の基板20は、例えば円形であってもよく、三角形、五角形、六角形、七角形、八角形等の多角形であってもよい。
図4(a)及び(b)は、本実施形態の粒子捕捉デバイスの一例を示す概略図である。図4(a)は正面断面図であり、図4(b)は上面図である。
図5(a)及び(b)は、本実施形態の粒子捕捉デバイスの一例を示す概略図である。図5(a)は正面断面図であり、図5(b)は上面図である。
上述した粒子捕捉デバイスは、複数連結されていてもよい。例えば、上述した粒子捕捉デバイス400又は500は、複数連結されて、6ウェルプレート型、12ウェルプレート型、24ウェルプレート型、48ウェルプレート型、96ウェルプレート型、384ウェルプレート型、1536ウェルプレート型等の形状に形成されていてもよい。特に、粒子として細胞を捕捉する場合、粒子捕捉デバイスのサイズは、実用上の観点から、広く細胞培養等に利用されている、SBS規格に準拠したサイズ、スライドグラスサイズ、又は、シャーレのサイズで作製することが好ましい。
1実施形態において、本発明は、上述した粒子捕捉デバイスの製造方法を提供する。本実施形態の製造方法は、第1の支持体上に、溶解可能な下地膜を形成し、該下地膜上に第1の硬化性樹脂組成物を塗布して第1の硬化性樹脂膜を形成し、該第1の硬化性樹脂膜に連通孔をパターニングして、連通孔がパターニングされた支持層を得る工程1と、前記支持層上に、第2の硬化性樹脂組成物を塗布して第2の硬化性樹脂膜を形成し、該第2の硬化性樹脂膜に凹部をパターニングして、凹部がパターニングされた第1の基板を得る工程2と、前記下地膜を溶解して、前記第1の支持体から前記第1の基板を剥離する工程3と、前記第1の基板と第2の基板とを接合させる工程4と、を含み、前記第1の基板と第2の基板との接合物が粒子捕捉デバイスである。
本工程では、例えば図6(a)に示すように、第1の支持体31上に、溶解可能な下地膜32を形成し、該下地膜32上に、第1の硬化性樹脂組成物を塗布して第1の硬化性樹脂膜10Bを形成し、第1の硬化性樹脂膜10Bを露光した後、現像して、図6(c)に示すような連通孔14がパターニングされた層10bを形成する。
本工程では、例えば図6(d)に示すように、層10b上に、第2の硬化性樹脂組成物を塗布して第2の硬化性樹脂膜10Aを形成し、該第2の硬化性樹脂膜10Aを露光した後、現像して、層10b上に、凹部13がパターニングされた第1の基板10を得る。
本工程では、例えば剥離剤(例えば、1-メチル-4-イソプロピルシクロヘキサン(p-Menthane))に基板ごと浸漬することにより、下地膜32を溶解し、第1の支持体31から第1の基板10を剥離する。
本工程では、上記工程で得られた図6(f)に示す第1の基板10及び図6(g)に示す第2の基板20を接合させる。接合する際には、層10bが第2の基板20と対向するように接合する。接合には、前記硬化性樹脂組成物を接着剤として用いてもよい。図6(f)に示すように、第2の基板20はピラー22を有していてもよい。
本工程では、例えば図6の(i)に示すように、第2の支持体20上に、第3の硬化性樹脂組成物を塗布して、第3の硬化性樹脂膜22Aを形成し、第3の硬化性樹脂膜22Aを露光して現像することにより、図6(g)に示すようなピラーパターン22を形成する。
1実施形態において、本発明は、上述した粒子捕捉デバイスの流入口に粒子を供給し、流出口から分散媒を流出させる工程を備える、粒子の捕捉方法を提供する。本実施形態の捕捉方法は、粒子を均一に捕捉する方法、均一に捕捉された粒子の製造方法等といいかえることができる。
(第1の基板の製造)
《連通孔パターニング》
シリコン基板上に、下地剤をスピンコータ―(1500rpm、20秒)で塗布し、ホットプレートにより90℃で1分間、120℃で3分間プリベークし、下地膜を形成した。
上記で得られた連通孔樹脂パターン上に、上記感光性樹脂組成物をスピンコータ―(1000rpm、20秒)で塗布し、ホットプレートにより90℃で5分間プリベークした。その後、ミラープロジェクションマスクアライナー(型式「MPA―600FA」、キャノン製)を用いてパターン露光(GHI線、60mJ)を行い、ホットプレートにより90℃で5分間露光後加熱を行った。その後、PGMEAを用いた浸漬法により、2分間の現像処理を行なった。次いで、現像後の樹脂パターンを、基板ごと、オーブンを用いて120℃で1分間ポストベークし、凹部パターンを得た。凹部は直径10μmの円筒状とした。
上記で得られた凹部がパターニングされた第1の基板を、剥離剤に浸漬し、上記下地膜を溶解することにより、シリコン基板から、連通孔樹脂パターン上に凹部パターンが形成された第1の基板を剥離した。
ガラス基板上に、上記感光性樹脂組成物をスピンコータ―(1000rpm、20秒)で塗布し、ホットプレートにより90℃で5分間プリベークした。その後、平行光露光機(伯東株式会社製、型番MAT-2501)を用いてパターン露光(ソフトコンタクト、GHI線、500mJ)を行い、ホットプレートにより90℃で5分間露光後加熱を行った。その後、PGMEAを用いた浸漬法により、2分間の現像処理を行った。次いで、現像後の樹脂パターンを基板ごと、オーブンを用いて120℃で1分間ポストベークし、第2の基板上に樹脂パターンを形成した。樹脂パターンは、後述する工程で第1の基板及び第2の基板を接合した場合に、第1の基板と第2の基板との間の距離(以下、「流路高さ」という場合がある。)を規定するものであり、第1の基板と第2の基板との間の距離が120μmとなる樹脂パターンを作製した。なお、流路高さが高い場合(例えば100μm以上)は、上記スピンコーターによる塗布工程を、目標の高さになるまで、繰り返して行った。
上記で得られた第2の基板樹脂パターン上部に接着剤を塗布し、35℃で1分間プレベークした。その後、上記で得られた第1の基板を連通孔パターンが下になるように第2の基板と接合し、平行光露光機(伯東株式会社製、型番MAT-2501)を用いて露光(ソフトコンタクト、GHI線、60mJ)を行い、ホットプレートにより35℃で3分間、90℃で1分間露光後加熱を行い、接着剤を硬化させることにより第1の基板と第2の基板を接合し、図1(b)に示す形状の実施例1の粒子捕捉デバイスを得た。
デバイス形状、凹部の直径、流路高さ、凹部連通孔開口面積の合計、流出口面積を表1に示すものにした以外は、実施例1と同様にして、実施例2~8、比較例1~2の粒子捕捉デバイスを作製した。
実施例1~11、比較例1~2の粒子捕捉デバイスに、培地に懸濁したNamalwa細胞を導入して捕捉した。Namalwa細胞は、予めCalcein-AM(同仁化学研究所製)で染色した。粒子捕捉デバイスに導入したNamalwa細胞の数は、それぞれ、粒子捕捉デバイスの凹部の数と同数にした。
Claims (5)
- 第1の基板と、前記第1の基板の一方側に平行に対向するように配置された第2の基板とを備え、
前記第1の基板は、前記第1の基板の他方側に開口し、粒子1個を捕捉可能な大きさを有する凹部を複数有しており、
前記凹部は、前記一方側と前記他方側とを連通し、前記粒子の分散媒が移動可能な大きさを有する連通孔を有しており、
前記第1の基板と前記第2の基板との間は、前記第1の基板の前記連通孔を前記分散媒の流入口とし、前記第1の基板の一方側の端部を前記分散媒の流出口とする流路を形成しており、
前記連通孔の開口面積の合計が1mm2以上10mm2未満であり、前記流出口における前記流路の断面積が、前記連通孔の開口面積の合計の0.8倍以上であるか、又は
前記連通孔の開口面積の合計が10mm2以上1000mm2以下であり、前記流出口における前記流路の断面積が、前記連通孔の開口面積の合計の0.1倍以上である、
粒子捕捉デバイス。 - 前記流出口における前記流路の断面積が前記連通孔の開口面積の合計よりも大きい、請求項1に記載の粒子捕捉デバイス。
- 前記第1の基板と前記第2の基板との間の距離が100μm以上である、請求項1又は2に記載の粒子捕捉デバイス。
- 前記粒子の直径が1~500μmである、請求項1~3のいずれか一項に記載の粒子捕捉デバイス。
- 第1の基板と、前記第1の基板の一方側に平行に対向するように配置された第2の基板とを備え、
前記第1の基板は、前記第1の基板の他方側に開口し、粒子1個を捕捉可能な大きさを有する凹部を複数有しており、
前記凹部は、前記一方側と前記他方側とを連通し、前記粒子の分散媒が移動可能な大きさを有する連通孔を有しており、
前記第1の基板と前記第2の基板との間は、前記第1の基板の前記連通孔を前記分散媒の流入口とし、前記第1の基板の一方側の端部を前記分散媒の流出口とする流路を形成しており、
前記連通孔の開口面積の合計が1mm2以上であり、
前記第1の基板と前記第2の基板との間の距離が100μm以上である、
粒子捕捉デバイス。
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