JP2003527972A - Modular microfluidic device containing sandwiched stencils - Google Patents

Abstract

(57) SUMMARY The present invention provides modular microfluidic devices or systems, and further provides methods for their manufacture. The microfluidic device includes first and second substrates (59, 60) and at least one stencil (58) sandwiched between the first and second substrates to form one or more sealed microstructures. ). The stencil is adhered to at least one of the first and second substrates by an adhesive (44). In a preferred embodiment, a plurality of sandwiched stencils are provided. Further, the first and second substrates are preferably substantially flat. These microfluidic devices can be rapidly prototyped with low machine tooling costs and can be easily assembled to form three-dimensional structures with complex microfluidic system structures.

Description

Detailed Description of the Invention

The present invention is directed to pending US Application No. 60 / filed Oct. 4, 1999.
Claims the right of No. 157,565.

FIELD OF THE INVENTION The present invention relates generally to modular microfluidic devices or components that can be combined with each other to form microfluidic devices.
In particular, the present invention relates to a modular microfluidic device including a laminated substrate and a stencil sandwiched between the substrates, and a method for manufacturing the same.

Prior Art There is increasing interest in the manufacture and use of microfluidic devices to obtain chemical or biological information. In particular, microfluidic devices enable complex biochemical reactions, which are achieved, for example, with liquids having very small volumes. These miniaturized devices increase reaction response time, minimize sample volume, and reduce reagent costs, among other advantages.

Traditionally, microfluidic devices have been constructed in a flat shape using techniques introduced from the silicon manufacturing industry. For example, Manz et al. Described earlier work (Trends in analytical chemistry (1990) 10 (5): 144-149; Advances in chromatography (1993) 33: 1-66). In these publications, microfluidic devices are constructed by using photolithography to form channels on a silicon or glass substrate and etching methods to remove material from the substrate to form the channels. ing. A cover plate is adhered to the top surface of the device to provide the closure. Also, a miniature pump or valve may be integrated (eg, internally) with the device.

In recent years, manufacturing methods have been developed that allow microfluidic devices to be constructed from plastic, silicon, or other polymeric materials. In one of the above methods,
First a negative mold is constructed and then plastic or silicone is poured inside or on said mold. The mold is a silicon wafer (eg, Duffy et al., Analytical Chemistry (1998) 70: 4974-4984; McCor.
Mick et al., Analytical Chemistry (1997) 69: 2626-2630) or by making conventional injection molding cavities for plastic devices. Some mold making machines have advanced technology for constructing ultra-fine molds. Components constructed using the LIGA method were developed at Karolsruhe Nuclear Research Center in Germany (see, for example, Schomburg et al., Micromechanical Microengineering Journal (1994) 4: 186-191), Micro.
Commercialized by oParts (Dortmund, Germany). Jenop
tik (Jena, Germany) also uses LIGA or hot-embossing methods. Also, a method of imprinting on polymethylmethacrylate (PMMA) is disclosed (eg, Martynova et al., Analytical Chemistry (1997) 69:47).
83-4789). However, these techniques do not provide rapid prototyping and manufacturing method flexibility. In addition, these techniques are limited to flat (ie two-dimensional or 2-D) microfluidic structures. In addition, the tool-up cost for these technologies is
Very expensive or expensive.

Gonzalez et al. Have reported microfluidics (sensors and actuators B (1
998) 49: 40-45). However, the microfluidic device described by Gonzalez et al. Is not truly modular as it is limited to interconnect systems to silicon wafers.

As mentioned above, there is a need for a modular microfluidic device that can be easily combined, constitutes a more complex microfluidic system, and is low cost and prototyping. There is also a need for modular, three-dimensional microfluidic devices that include various components (eg, valves, filters, etc.).

SUMMARY OF THE INVENTION The present invention addresses the aforementioned needs and provides additional advantages over conventional microfluidic technologies. The modular approach to the microfluidic device of the present invention and the included versatility and low cost manufacturing method make it “generic”.
) "Microfluidic devices consisting of device components can be constructed and assembled and combined easily and effectively to meet a wide variety of design considerations and requirements. Due to the modular design, expensive custom micro Eliminates the need to design and manufacture fluid systems.

One object of the present invention is to provide a cheap, modular and modular microfluidic device. Another object is to provide a microfluidic device that can be rapidly prototyped with minimal machine tool equipment costs. The manufacturing cost of the microfluidic device according to the present invention is relatively low regardless of whether the product volume is large or small.

Another object of the present invention is to provide a modular system of microfluidic components that can be combined in various configurations to form microfluidic devices. In this way, prototyping and manufacturing can be achieved in a very rapid manner, which allows the bulk construction of a complete set of generic "assembled blocks". And, these components and devices can be combined in various ways to form the desired microfluidic system.

Yet another object of the present invention is to provide an inexpensive manufacturing method for manufacturing convex or concave molds for constructing microfluidic replicates.

[0012] Yet another object of the present invention is to provide "built-in" (ie, integral) electronic components within a microfluidic device. In particular, electrodes may be placed inside the channels or chambers of the microfluidic device, for example.
These electrodes are preferably used in particular for electrokinetic flow, electrophoresis, electrochemical detection, impedance or temperature detection.

Another object of the invention is to provide a microfluidic device having a valve for controlling or manipulating a fluid.

Another object of the present invention is to provide a microfluidic device having a microstructure capable of filtering a small amount of fluid, especially a fluid containing biomolecules such as nucleic acids or proteins.

Yet another object of the present invention is, but not limited to, a microfluidic that is chemically compatible with or capable of use with a large array of liquid reagents or solutions containing organic solvents such as acetonitrile. Is to provide the device.

These and other objects are provided by the present invention. In a preferred embodiment, the microfluidic device comprises a first and a second substrate, and is sealed with one or more by placing (eg, sandwiching) at least one stencil between the first and second substrates. Microstructures are formed in between. The stencil is adhered to at least one of the first and second substrates with an adhesive. In the preferred embodiment, there are multiple sandwiched stencils. Preferably, the first and second substrates are substantially flat and have planar surfaces that are complementary to each other to ensure sealing of the microstructures therebetween. The first and second substrates are preferably made of Mylar (registered trademark), FR-4, polyester, glass, acrylic resin, polycarbonate and fiber glass.

The adhesive may be a rubber-based adhesive, an acrylic resin-based adhesive, or a resin-based adhesive. In a preferred embodiment, the stencil preferably has an adhesive. In most preferred embodiments, the stencil comprises an adhesive tape that is a one-sided (ie, adhesive on one side) or double-sided (ie, adhesive on both sides) tape. Any adhesive tape may be used, including in particular commercially available adhesive tapes. Types of such adhesive tapes include, but are not limited to, pressure sensitive tapes, temperature activated (eg, heat activated) tapes, chemically activated (eg, epoxy resin 2 parts) tapes and photoactivated (eg, UV activated) tapes. To be The adhesive tape preferably comprises a backing material selected from the group consisting of Mylar®, nylon and polyester to support the adhesive. In another embodiment, the stencil and at least one of the first and second substrates are preferably ultrasonically welded to each other.

The stencil is particularly preferably composed of polymers, paper, fabrics and foil foils. The stencil may be Mylar (registered trademark), polyester, polyimide, vinyl, acrylic resin, polycarbonate, Teflon (registered trademark), Kapton (registered trademark), polyurethane, polyethylene, polypropylene, polyvinylidene fluoride, It may comprise a polymer selected from the group consisting of polytetrafluoroethylene, nylon, polyether sulfone, acetal copolymer, polysulfone, polyphenyl sulfone, ABS, polyvinylidene fluoride, polyphenylene oxide, and derivatives thereof. preferable.
In one embodiment, the stencil preferably comprises a fluorinated polymer commonly known to be chemically resistant. The stencil is preferably made of an elastomeric material such as rubber, viton and silicone. The stencil used in the present invention is preferably die-cut by, for example, an automatically controlled rotary die-cutting machine.

The microfluidic device according to the present invention preferably has a sealant coating on at least a part of one or more of the stencil, the first substrate and the second substrate. The sealant coating adheres the substrate and stencil to each other,
It is preferred to help seal the microstructures formed therebetween.
The sealant coating is preferably made of a silicone material. In addition, the sealant coating is Teflon (registered trademark), Avatrel (registered trademark), Liquin (registered trademark), perfluorocarbons, fluorinated thermoplastics, polyvinylidene fluoride, acrylic resin, It is preferably composed of one or more materials selected from waxes, epoxy resins, solders, polymers, paints, oils and varnishes. The sealant coating is a spin-deposition,
It is preferably formed by a number of different methods including spraying and dipping.

The microfluidic device preferably comprises one or more microstructures of channels or chambers. In one application, the microstructures are at least partially filled with a fill material, such as a filter material. The filter material comprises a wide variety of materials that allow specific or arbitrary filtration with various dimensional parameters. Any kind of chemical or biological or size-exclusion filter material can be used. In one embodiment, the filter material is selected from the group consisting of polycarbonate, acrylic resin, acrylamide, polyurethane, polyethylene, polypropylene, polyvinylidene fluoride, polytetrafluoroethylene, naphion, nylon, polyethersulfone. It is preferably one. The filter material may also be selected from the group consisting of agarose, alginate, starch and carrageenan. Preferably, the filter material is Sephadex (R), Sephakill (S).
ephacil) (registered trademark) or hydroxyapatite. In a preferred embodiment, the filling material is preferably formed by silk screen, which reduces manufacturing time and costs. The filling material may be formed by lithography. Further, the filling material is preferably formed by a pick-and-place method known in the semiconductor manufacturing industry.

In one embodiment, the microfluidic device comprises at least one substrate having one or more holes that allow fluid to move between the one or more substrates. Also, the microfluidic device may further comprise one or more valves of various designs. Some valve shapes and components are described below. The microfluidic device may be used to divide a liquid sample into multiple samples. In one embodiment, such sample division is achieved by using microstructures having one or more fork-shaped channels. Each channel preferably has one or more constrictions for controlling the fluid flowing therein.

The microfluidic device may further comprise at least one electrode.
The electrodes are used to detect or measure the electrical properties of the fluid. Alternatively, the electrodes may be for promoting electrophoretic or electrokinetic flow of fluids.

The microfluidic device may also be combined with external optical spectroscopy, response and rectification. Thus, in certain embodiments, at least a portion of at least one of the substrates of the microfluidic device is one that allows passage of an optical signal (eg, is sufficiently optically transparent). Is preferred.

Microfluidic devices according to the present invention are preferably stacked to form a three-dimensional device or system. Therefore, in certain embodiments, the microfluidic device may further be provided with one or more additional substrates that are sealed and secured. This additional substrate is preferably stacked or stacked so that the multiple layers of microstructures are properly positioned to function for the desired application. The one or more additional substrates may comprise an electronic circuit board having microstructures on its surface. The utility of electronic circuit boards in constructing microfluidic devices is disclosed in pending US Application No. ______ (Attorney Case No. ______), the disclosure of which is hereby incorporated by reference. Be incorporated. In addition, a microfluidic system including a plurality of microfluidic devices is preferably configured by arranging at least two microfluidic devices so that fluids can move with respect to each other. The microfluidic system is
It is preferably prepared by stacking two or more microfluidic devices to form a three-dimensional microfluidic system.

The present invention also provides a manufacturing method for manufacturing a microfluidic device. The manufacturing method includes: (a) providing a first substrate; (b) stacking one or more panels of a stencil array on the first substrate; and (c) on the one or more panels. And laminating a second substrate to form a plurality of microstructures therebetween. Preferably, at least one of the first and second substrates has one or more holes. It is also preferred that at least one of the panels is positioned on at least one of the first and second substrates so that the holes allow fluid to move between the microstructures. Such positioning is preferably provided by peg-hole alignment. In addition, the present invention preferably provides, in an embodiment, a microfluidic device prepared by the manufacturing method described above.

The present invention also provides a mold prepared for using at least a portion of a stencil as a mold for forming a mold. The mold is preferably made of a silicon material. A microfluidic device containing microstructures may be prepared for use with the mold.

Definitions As used herein, the term “microfluidic” refers to a microstructure having one or more dimensions less than or equal to 500 microns, as well as components, devices comprising such microstructures. And by reference to the system. The microfluidic device of the present invention may be planar (i.e., approximately two-dimensional, or 2-D) or three-dimensional (3-D). In addition,
Such devices can be constructed by using some of the materials described herein, combinations of the above materials, and similar or equivalent materials.

The term “microstructure” as used herein refers to a microfluidic structure disposed on one or more substrates used to fabricate the microfluidic device of the present invention. The term includes any structure capable of supporting a fluid (including, but not limited to, channels or chambers) (eg, a microstructure capable of passing, storing, and directing an inflowing or inflowing fluid). The boundaries of the microstructure are formed by the contours of the excised portion of the sandwiched stencil.

The term “seal” as used herein refers to a microstructure that has a sufficiently low rate of unintended leakage and / or volume under a given flow, fluid type and pressure conditions. The term also includes microstructures having one or more holes therein for the passage of fluid.

The term “adhesive” as used herein is effective in adhering the various stencil and / or substrate layers of the microfluidic device of the present invention to each other and forming a sealed microstructure therebetween. Refers to any chemical substance that has an adhesive property of

The term “channel” or “chamber” as used herein is limited to elongated shapes whose transverse or longitudinal dimensions are much greater than their thickness, depth or cross-sectional dimensions. Not a thing. Rather, the term is intended to include cavities or tunnels of the desired shape or profile through which liquid is guided or passed. Such fluid cavities include, for example, a flow-through channel through which fluid can pass continuously, or, alternatively, a chamber for holding a quantity of separated fluid. A "channel" or "chamber" may be filled or contain internal structures, for example, valves, filters, and similar or equivalent components or materials.

As used herein, the term “stencil” refers to a material, preferably a generally flat material, that has one or more portions of various shapes or orientations cut or removed. The profile of the excised or removed portion consists of the lateral boundaries of the microstructure formed on the stencil sandwiched between the substrates.

The microfluidic devices described herein are “generic” in that they are modular and easily reconfigurable or adaptable to any shape. These devices can use a variety of pump or valve mechanisms, such as pressure, peristaltic pumping, electrokinetic flow, electrophoretic, vacuum, and the like. In addition, the microfluidic device of the invention can be used in combination with optical detection (eg, fluorescence, phosphorescence, luminescence, absorption, colorimetric), electrochemical detection, and any of a variety of suitable detection methods. . Suitable detection methods depend on the device geometry and compositional components. Selection of the appropriate detection method for a given application will be apparent to those of skill in the art.

DETAILED DESCRIPTION OF THE INVENTION Referring to FIG. 1, a microfluidic device is constructed by sandwiching a stencil between two substrates. Referring to FIG. 1A, a channel 57 is constructed by sandwiching a stencil 58 between two substrates, shown by a bottom substrate 59 and a top substrate 60. In the above embodiment, the inlet and outlet holes 61
Both are positioned within the top substrate 60. The assembled device is shown in FIG. 1B. The inlet and outlet holes preferably communicate either outside of the device or to adjacent stencil and / or substrate layers. In other embodiments (not shown), the stencil layers may be laminated directly to each other without being sandwiched between the substrates.

Microstructures (eg, channels or chambers) may be formed in the stencil before or after being placed on the support substrate. In a preferred embodiment, the stencil is formed by cutting or removing a portion of the stencil material of suitable shape and orientation prior to being placed on the substrate. For example,
The stencil may be excised, preferably using an automatically controlled die cutter. Alternatively, the ablation may be accomplished using a laser cutter. In a preferred embodiment, the stencil is ablated using a computer controlled die cutter or laser cutter. In another preferred embodiment, the ablation comprises a rotary cutter, a printer press, or any high throughput auto-aligning device.
equipment) is used. Such a device is called a converter.

In a preferred embodiment, the stencil comprises a tape having adhesiveness on one side or both sides. A portion of the tape (of desired shape and size) is preferably cut or removed to form, for example, a channel or chamber. Next, the tape stencil is placed on a supporting substrate or between the substrates. In one embodiment, the stencil layers are laminated together. The thickness or height of the channels can be varied by simply changing the thickness of the stencil (eg, tape carrier or adhesive layer thereon).

Various types of tape are available in the above embodiments. The type of adhesive, as well as the thickness and components of the underlying carrier, can be varied to suit the application. Suitable tapes for use in the present invention have various methods of curing or activating and, among other types of tapes, pressure sensitive tapes,
It is preferred to include heat activated tapes, chemically activated tapes, and light activated tapes. For example,
A variety of adhesives are available, including rubber-based adhesives, acrylic resin-based adhesives, and gum-based adhesives. It also carries the adhesiveness (carry)
There are numerous materials used for this. One example of a suitable tape carrier material is Mylar®, which consists of polyester and nylon. The thickness of the carrier can be changed.

In a preferred embodiment, a probe is used to form the channel or chamber of the stencil. In one embodiment,
The probe is, for example, a cutting device mounted on a computer-controlled plotter. The probe selectively removes from the material features that form lateral boundaries of microstructures (eg, channels or chambers) to form stencils. In one embodiment, a heat probe is used to selectively melt or anneal the heat activated adhesive to form microstructures. In other embodiments, ultrasonic welding may be used to form microstructures in layered stencils. These layers may be "melted" together by using ultrasonic welding.

In a preferred embodiment, the material forming the stencil is applied on the substrate inside a predetermined desired area using a silk screen. And
The material is "cured" to form channels and / or chambers. In the examples, activatable or curable polymers are used as stencil materials. In another embodiment, paint or ink is used as the material. One example is the Genesis Artist Colors (Indinap).
Thick Medium heatset available from olis, IN)
-set) Acrylic resin is used. In another embodiment, the entire surface of one of the substrates is covered with the stencil material. Then, it is preferable that the stencil is cured in the remaining region and the remaining material is removed. In this embodiment, a curable epoxy resin material may be used. In a more preferred embodiment, the epoxy resin is a UV curable epoxy resin. Alternatively, two parts of epoxy resin may be used, in which case the first part is patterned in place and then the entire device is dipped into the second part which adheres the stencil material within the predetermined area. It

The chemistry of the stencil material, and thus the chemistry of the microstructures, can be “tuned” for a particular application. The stencil material preferably has hydrophilicity, hydrophobicity or ionicity. The stencil material is flexible. In various preferred embodiments, the stencil material is selected from the group consisting of vinyl, filter material, paper or fabric, foil foil and foam or foam sheet. In another preferred embodiment, the stencil material is formed of polymeric material. Suitable polymers include, but are not limited to, polycarbonate, acrylics, polyurethane, polyethylene including high density polyethylene (HDPE) or ultra high molecular weight polyethylene (UHMW), polypropylene (PP), polyvinylidene fluoride (PVDF), polytetra. Fluoroethylene (PTFE), nylon, polyether sulfone (PES),
Includes acetal copolymers, polyester imidas, polysulfones, polyphenyl sulfones, ABS, polyvinylidene fluorides, polyphenylene oxides, and derivatives thereof. In a particularly preferred embodiment, the polymer is a fluorinated polymer. This is because fluorinated polymers have excellent resistance to aggressive solvents such as organic solvents.

In a preferred embodiment, the stencil material is a flexible or elastomeric material that enables a valve or pump mechanism, such as silicone, viton, or rubber. When pressure or mechanical force is applied to the flexible layer, the material flexes or occludes the channels above or below it.

In a preferred embodiment, the sealant coating provides for coating or sealing the microstructure. Referring to FIG. 2, at least the stencil 3
3 and / or a portion of the surface of the substrate 30 is coated with a layer of sealant coating material 38. A (preferably substantially flat) cover plate substrate 39 is overlaid on the stencil 33 to "cap" or complete the microstructures 35 formed between the substrates 30 and 39. In FIG. 2C, the cover plate substrate 39 is uncoated. In FIG. 2D, the cover plate substrate 39 is coated with a sealant coating material 41, which may be similar or different than the sealant coating material 38. Referring to FIG. 3, a small amount of epoxy resin 44 has been added to help bond the cover plate substrate 39, substrate 30 and stencil 33 together. The epoxy resin 44 is added before or after the sealant coating material 38 is cured. In other preferred embodiments, the layers of the device may be mechanically held together. For example, a gasket may be used to help seal the microstructure.

A number of suitable sealant coating materials having various desired properties may be used. The sealant coating material has chemical and / or biological properties and is either hydrophobic or hydrophilic, depending on the application. Solid, liquid, gel or powder form, or combinations thereof can be used. Materials capable of conducting surface charges may be used, as well as materials of neutral nature. Examples of suitable coating materials for use in the present invention include Teflon (R), Liquin (R)
, Abatrel®, silicone, silicone mixture, epoxy resin (including solder mask), glue, liquid polymer, polymerizable dispersion, plastic, liquid acrylic resin, paint, metal, oil, wax, photo Resist, varnish,
Includes solder and glass.

In a preferred embodiment, the sealant coating material is a polymer such as polyethylene glycol or cyanoacrylate. In another preferred embodiment, the coating material has biological properties. Conveniently, in various applications, the biological coating material can be used to either promote or prevent the adhesion of the material. In certain embodiments, biological coating materials (eg, ligands) that specifically bind to a given biological material are preferred. Examples of biological coating materials include proteins, antibodies, lipids, cells, tissues, nucleic acids and peptides. More specific examples include avidin, streptavidin, polylysine and enzymes. In certain embodiments, the coating material is used to selectively bond the listed example materials.

In another preferred embodiment, the sealant coating material is a fluorinated polymer. Fluorinated polymers have excellent resistance to various solvents or chemicals, including organic solvents. In particular, as an example, Teflon (registered trademark)
, Abatrel®, polyvinylidene fluoride (PVDF), THV fluorinated thermoplastic (Dyneon, StPaulMN), Hostaflon TF
5035 (Dyneon) can be mentioned.

Various sealant coating materials are formed using a number of techniques. In a preferred embodiment, a spinner or rotator is used to spin-coat on a given substrate and / or stencil. In particular, a suitable amount of sealant coating material is positioned on the substrate or stencil and the entire substrate or stencil is rotated to create a globally uniform sealant coating layer. In a preferred embodiment, the rotational speed is between about 10 revolutions per minute (rpm) and about 100,000 rpm.
More preferably, the rotation speed is between about 500-20,000 rpm, most preferably about 1,000-20,000 rpm. Multiple spin-deposition cycles may be used to create a coat thickness machine.

Alternatively, the sealant coating material may be formed by spraying the surface with the sealant coating material. For example, the sealant coating material is ultrasonically sprayed through a nozzle or other orifice.
) Will be done. In one embodiment, a colloidal dispersion of sealant coating material is provided at a concentration adjusted to give a layer of desired thickness when sprayed onto a surface. In another embodiment, the sealant coating material is sprayed directly onto the surface. In yet another embodiment, the sealant coating material is dissolved in a suitable solvent and then sprayed onto the surface. That is, as the solvent evaporates, the sealant coating material remains behind forming a coating layer. The sealant coating material may also be applied by dipping the substrate and / or stencil into a volume of sealant coating material. By dipping once, a coating having a predetermined thickness can be formed. It may be dipped several times to form a thicker coating. Alternatively, the sealant coating material may be brushed onto the surface. By the spraying method as described above, the sealant coating material is directly formed as a colloidal dispersion or as a material dissolved in a solvent. In another preferred embodiment, the sealant coating material is stamped on the surface.
) Will be done.

In a more preferred embodiment, the sealant coating material is patterned (eg, by silkscreening) on the surface. In this embodiment,
The sealant coating material is used to coat only selected areas of the surface as formed by a silk screen mask. In another preferred embodiment, photoresist patterning is used to achieve liftoff or etch patterning. The photoresist is then removed, leaving only the coating on the predetermined areas of the surface. This process is repeated as desired or needed using different photoresist patterns and coating materials.

In other embodiments, various thin film deposition methods are used to form the sealant coating material. Such techniques include, but are not limited to, thermal evaporat
ion), e-beam evaporation, sputtering, chemical vapor deposition, and laser vapor deposition. These and other thin film forming methods are known to those skilled in the art. In addition, plating methods are used to form the sealant coating material. Such plating methods include, but are not limited to, electroplating or chemical plating of metallic materials.

The thickness of the sealant coating is important in certain embodiments. Preferably, the thickness of said coating chemically protects the underlying surface and / or
It need only be sufficient to bond or seal adjacent substrates and / or stencils. A potential problem with overly thick coatings is obstruction or blockage of microstructures, which can obstruct or obstruct the fluid flowing within it.

If the sealant coating material alone cannot provide the adhesive function, a thinner coating is used. In practical use, it is preferable when there is a molecular layer (or monolayer). In a preferred embodiment, the sealant coating is a self-assembled monolayer of alkanethiol that is particularly easily formed on a metal surface such as gold.
Other similar thiols may be used. In another preferred embodiment, a silanization reaction may be used to coat the substrate. Silanization is known to minimize the adhesion of certain biological materials such as nucleic acids or peptides. In yet another preferred embodiment, the microstructures are coated with a bilayer or multilayer of lipids. In certain embodiments, these molecular monolayers are terminated with biological molecules that are used to bind the molecules within the solution. An example thereof is alkanethiol coated with nucleic acid or silane coated with protein.

In some embodiments, a second mechanism may be used to help seal the substrate and / or stencil together. In some embodiments, the layers are mechanically held together. Examples include the use of nuts and bolts, tightly fitting pegs and holes, epoxy resin, BLU-TEK®, or external clamps. Alternatively, pressure or vacuum may be used to achieve this mechanical bonding or sealing.

Prior to the coating step, it is necessary to adjust the tack of the sealant coating material. To obtain the desired tack, the sealant coating material needs to be diluted or diluted with another solvent or chemical. Alternatively, the sealant coating materials may be heated prior to forming them to change their tackiness. Appropriate stickiness adjustments will be apparent to those skilled in the art.

The coated substrate and stencil are preferably cleaned prior to the coating and bonding steps. Examples of cleaning materials include soaps, surfactants, surfactants, organic solvents and Freon®.

In another preferred embodiment, a flexible sealant coating material may be used on a given layer of the device to implement a valve or pump mechanism. A preferred flexible sealant coating material is silicone rubber. Pressure or mechanical force may be applied to the flexible layer to cause the material to deflect or occlude channels located above or below it.

Three-dimensional structures may be formed using stencils that form channels and / or chambers. Referring to FIG. 4, a microfluidic device including a filter is constructed. Stencil 64 with T-shaped channel 62 made of double-sided tape
It has been resected inside. The stencil 64 is glued to a top plate 66 having two inlets 68. The stencil support layer 70 is adhered to the bottom surface of the stencil 64. The support substrate layer 70 has holes 72 communicating with the adjacent stencil layer 74. The stencil layer 74 is composed of a double-sided tape which is formed by cutting and removing the filter channel 76 and the filter chamber 78. The filter chamber 78 is filled with any kind of suitable filter material, depending on the application (see examples below). The bottom substrate 80 is adhered to the stencil layer 74 with the filter components. An outlet hole 81 is formed in the bottom substrate 80. In actual use, the sample is injected into one of the inlet holes 68 and the reagent is injected into the other of the inlet holes 68. The sample and reagent mix at the confluence of the T-shaped channels and pass down to the adjacent layer. The filter material in the filter chamber 78 captures unwanted material and the purified material passes through the outlet. FIG. 4B shows the assembled device.

Diagrams of different 3-D microfluidic devices are shown in FIG. In this embodiment, the upper three layers are similar to the upper three layers shown in FIG. However, instead of constructing a channel for the filter and filling the channel with a suitable filter material, layer 84 itself consists of the filter material. The bottom layer 80 having the outlet holes 81 is similar to that shown in FIG. FIG. 5B shows the assembled device.

Methods of forming holes include, but are not limited to, mechanical drilling, laser drilling, chemical etching, plasma etching and punching punching. Alternatively, the components of the device may consist of injection molded parts with integrally formed or equipped inlet / outlet holes. Other techniques known to those skilled in the art for forming through holes may be used.

In one embodiment, the sealant coating material may be chemically bonded to the underlying substrate or an adjacent layer. Alternatively, non-covalent bonds (non-
Covalent chemical interaction) may be used to hold the substrates together. The stencil material may be melted on the underlying substrate or may be adhered by an adhesive or other technique, such as heating. In other embodiments, the stencil may be mechanically pressed against a lower or adjacent substrate.

The electronic circuit board having microstructures on its surface may constitute one or more layers of the microfluidic device. Constructing a microfluidic device using an electronic circuit board type substrate and another substrate having a metal laminate is for the purpose of pending US application number ______ (Attorney case number ______). is there. The disclosure of which is incorporated herein by reference.

In a preferred embodiment, the microstructures are preferably filled with various types of packing materials, including reagents or catalysts. In certain embodiments, these filler materials are preferably used to achieve useful chemical and / or biological reactions. In a preferred embodiment, the packing material is a filter useful for separating and / or purifying the material. These filters are preferably chemical or biological filters or size-exclusion filters. These filters may trap unwanted material, or, in the alternative, trap the desired material and later extract it. The filling material may be hydrophobic or hydrophilic, or charged or neutral. Also,
The filling material may be porous with various pore sizes. In a preferred embodiment, the fill material used to fill the channel or chamber is polymerizable. Non-limiting examples include polycarbonate, acrylic resin, polyurethane, high density polyethylene (HDPE), ultra high molecular weight polyethylene (UHMW), polypropylene (PP), polyvinylidene fluoride (P
VDF), polytetrafluoroethylene (PTFE), naphion,
Examples thereof include nylon and polyether sulfone (PES). In a preferred embodiment, the material used to fill the channel is a carbohydrate such as agarose, alginate, starch, or carrageenan. Further, the polymer may be an electroactive polymer. In a preferred embodiment, the packing material is silica gel. In another preferred embodiment, the filling material is Sephadex® or Sephakill®. In another preferred embodiment, the material used to fill the channels is acrylamide or agarose. In another preferred embodiment, the material used to fill the channels is hydroxyapatite.

In a preferred embodiment, the filling material used to fill the channels and / or chambers is a biological material. As an example, without limitation,
Included are binding proteins, antibodies, antigens, lectins, enzymes, lipids, and any molecule that interacts specifically or non-specifically with one or more species in a fluid.

In a preferred embodiment, the filling material is made of powder such as charcoal or porous beads. In another preferred embodiment, the filling material is
A reagent that is activated during use of the device. Two examples of this are soluble reagents or catalysts.

In a preferred embodiment, the filling material is a paper filter. The paper filter is a commercially available material that has been chemically modified to achieve specific functions such as capturing material or filtering various materials.

In a preferred embodiment, the material disposed within the microstructure achieves a useful biological or chemical action. One example is a solid buffer material that can be used to buffer the injected sample once. Other materials include lysis buffers for lysing cells or solid antibodies. Also, the catalyst may be placed inside the device.

In a preferred embodiment, the filling material consists of a single component already formed before being placed inside the microstructure. Alternatively, the material may be formed from multiple components so that it can be placed separately in the channel.
In this case, once placed in the channel, the material acts to form the desired fill material. Such curing can be accomplished by a variety of methods and is preferably spontaneous or caused by other effects such as light, heat, catalysts, solvents, drying and the like.

In one embodiment, the fill material is disposed within the microstructure during the manufacturing process. In this method, high throughput methods may be used to fill the channels. In one embodiment, high throughput pick-and-place equipment, such as used in the electronics industry, is used to deposit filter material.

In one embodiment, the filling material is patterned within the microstructures, for example by silk-screening the material in channels or by using lithography or by mechanically placing the material. To be done.
Referring to FIG. 6A, an “empty” microfluidic device 90 having two filter chambers 91, 92 is shown. Filter material into the filter chamber 9
Two silk screens 93, 94 have been made for placement within 1, 92. The screen and stencil are formed using materials that are compatible with the screened filter. Various screen materials and stencils can be used. One of the screens is positioned, for example, above the device, and the first filter material 95 is screened onto the device via the screen 93. The second filter material 96 is then screened onto the device via screen 94 in a manner similar to how the "final" device 98 is manufactured. The tack and thickness of the screened material are preferably adjusted to properly fill the filter chamber or other filter area. In addition, the total amount of material screened should be adjusted to properly fill the chamber.

In a preferred embodiment, all panels of the device are preferably coated simultaneously. Referring to FIG. 6B, the stencil is constructed on the panel 111. The preferred panel size is about 18 "to 24". However, other panel dimensions may be used. A collimation mark 114 is placed on the panel for visual or optical positioning. The holes placed in the stencil are
Used to position stencils on various machines used during the device manufacturing process. Silk screens 112, 113 of filter material are positioned on the device on the panel 111. All the filter chambers (reference numeral 11 indicating two types of filter chambers in the panel 111 are determined by one positioning).
5, 116) can be filled simultaneously. Finally, a substrate and / or additional stencils may be added to complete the microfluidic system.

In a preferred embodiment, one or more layers of the microfluidic device according to the invention may be used as a valve. Referring to FIG. 7, a multi-layer purification device is constructed. Inlet holes 100 and outlet holes 101 are formed in the top substrate layer 102. In one embodiment, the substrate layer is a flake of acrylic resin, glass or polycarbonate. The stencil forms the lower adjacent layer. Within this stencil, a filter chamber 104 is constructed and filled with a suitable filter material (not shown), for example by silk screen. Referring again to FIG. 7, a T-shape is formed, and one arm of the T-shape is formed in the chamber 1.
05, and the other arm is the channel 103. A hole 106 is located at the end of the chamber 105. The channel 103 communicates with the exit hole 101 in the top substrate layer 102. The bottom substrate layer 110 of the stack is made of a filter material that allows gas to flow but that closes when in contact with liquid. An example of such a filter material is X-7744, Porex Technologies.
7 μm pore size T3 sheets and Goretex® type materials commercially available from (Fairburn, GA).
In actual use, the sample is injected into the inlet hole 100. The sample passes through the filter material in the chamber 104 and into the “wide” chamber 105 and the “large” hole 106, injecting a sufficient volume of sample to fill the chamber and holes. . The liquid then flows inside the “narrow” channel 103 and exits the exit hole 1
It flows from 01 to the outside. A number of mechanisms may be used to initially preferentially flow the liquid inside the larger chamber 105. Capillary force is beneficial. Alternatively, the channels may be coated with a material that directs preferential flow.

Other valve mechanisms may be used when pressure and / or vacuum is applied to the inlet and outlet holes from an external power source. The flow rate of the fluid may be controlled by changing the external pressure or the flow rate of the input mechanism. One example is the use of syringes or peristaltic pumps. Alternatively, lateral micro-necking may be formed in the channel to limit the flow rate. Alternatively, a vertical waist may be added inside the channel. Also, these constrictions may be used to facilitate mixing. Alternatively, the flow rate in the network of channels may be limited or controlled by passing the liquid through a channel or part thereof coated with a compound that has a hydrophobic character compared to the rest of the network.

In a preferred embodiment, the microfluidic device is used to concentrate a sample. The device is configured such that the volume of the wide channels / chambers and large holes is approximately 2-100,000 times larger than the remaining filter chamber and channel volumes. Inject or flush large samples multiple times (wa
sh) is possible. Then, a very small volume of eluent may be added to remove sample adhering to the filter material in the filter chamber 104.

In another preferred embodiment, the valve is constructed by changing the shape of the channel itself. Referring to FIG. 8B, the device has a plurality of channel dividers or branches 120. Downstream of each split, the channel is constricted so that local capillary forces increase in that region (see 122 in Figure 8B). You may further comprise a division part on the downstream side of the constricted region. Referring to FIG. 8B, the fluid generally passes preferentially downstream of one of the two channels of the split so that the sample enters the split or base. However, once the fluid reaches the constricted region, the other of the two channels is filled because the capillary force in this region is small compared to the capillary force in the constricted portion. When both channels are filled, the fluid will pass through the constriction region and into the next channel. Using this approach, the sample can be accurately distributed in approximately equal parts. A 3D device utilizing this design is
Allows the sample to be divided into many compartments.

In another preferred embodiment, the valve is constructed by modifying the outlet holes for each channel. Referring to FIG. 9C, the microfluidic device is configured to have a large main channel 125 with a small branch channel 126. The main channel 125 ends in a channel 127 that is narrower than the channels 125 and 126. A cover plate substrate 128 with inlet and outlet holes positioned at one end of each of the channels described above forms the top substrate layer. When the fluid enters the main inlet hole 129, it first flows into the main channel 1
25 toward the downstream side. When the fluid reaches the confluence of the channels 127, the capillary force ensures that the fluid flows through the channels 126 downstream.

The surface chemistries of the various channels may be modified to achieve similar goals. In one embodiment, one end of the large channel is coated with a hydrophobic material and the rest of the channel is hydrophilic. When water enters the large channel, it first flows downstream through the large channel. Water is a hydrophobic end group (term
Upon reaching the region of the channels derivatized by the inal group or coating, the surface tension causes the remaining water to be small and the hydrophilic channels to flow downstream. Similar techniques may be used for organic solvents.

In multi-layer microfluidic devices, a consideration is the positioning of the layers. Therefore, it is preferable to use an automated technique for this positioning. In addition, these devices are preferably configured in parallel so that a single positioning can achieve the configuration of multiple devices. In a preferred embodiment, a peg-and-hole method may be used to position the layers. Referring to FIG. 10, the manifold 130 is configured with a dwell 132 having a particular diameter. The stencil panel 134 allows the manifold 13 to
It is located on 0. Here, the positioning is achieved by positioning the hole 135 in the dwell 132. Adjacent layer 136 is positioned over panel 134. Similarly, the positioning is accomplished by positioning the hole 137 in the dwell 132. Also, such a manifold may be used to aid in silkscreening.

In another preferred embodiment, edge alignment is used. Once the edges of the device are positioned, the channels and chambers can be automatically positioned and the device can be cut to size. Positioning may be accomplished mechanically, optically, magnetically, or otherwise.

In a preferred embodiment, the panels of the device are simultaneously positioned and processed. The completed panel structure is then cut or sectioned into individual devices. In a preferred embodiment, the layers of components are scored in the device portion prior to assembly.

In another preferred embodiment, the layers of the device are transported in close proximity, reorganized and pressed together. In these and other embodiments, it is important to control the humidity and temperature of the environment surrounding the layer. Often, the coating material or adhesive causes local static interactions between the layers, making positioning difficult. In an additional embodiment, the layer may be positioned in water to minimize the static forces. In other embodiments, a small amount of soap may be added to the water to prevent immediate adhesion if slight misfits occur. In other embodiments, the temperature of the local environment around the layer may be controlled for positioning.

In another embodiment, a press with alignment blocks is configured on both the top plate and the bottom plate. The layers of the device are positioned in the bottom alignment jig and the next layer is positioned in the top alignment jig. The device is then bonded together with an automated press. When the press section is expanded, the two layers remain attached to the bottom block. Adjacent layers may be added in a similar manner.

A converter or die cut press may be used to construct or stack the stencil. For example, the material forming each stencil layer may be loaded into the machine in roll form. A mechanical die punch is formed in each layer. The material is rolled out, punched and formed into flakes from each other by an automated method. Companies such as Acutek (Los Angels, CA) provide services that use converters. In a similar way, it is possible to use a rolling system to fill the channels and chambers. The roll of device is preferably pulled across the alignment block and the fill material is added as described above. Again, positioning is preferably accomplished by using a peg-hole alignment block. However, other positioning methods may be used, including optical positioning or precision positioning.

In a preferred embodiment of the invention, electrodes may be placed in the channels or chambers, for example to achieve detection and / or activation effects. Examples include electrophoresis, electrokinetic flow, electrochemical detection, impedance detection, capacitive detection, heating, and measurement of current or voltage. In other embodiments,
Interesting structures have been created to guide other phenomena. For example, a metal wire located within the microstructure may be energized to conduct heating inside the microstructure. A thermocouple may be constructed inside the microstructure using the metal wire to detect temperature changes. Calorimetry may be achieved by this method. In addition, it is possible to induce a magnetic field in a similar way. This magnetic field can be used to detect certain phenomena or induced flows using magnetic particles.

In another preferred embodiment, stencils are not used as the fluidic device itself, but they (or part of them) are used as shapes to form convex or concave molds. Various mold materials such as moldable polycarbonate or various silicones (see, eg, Duffy et al.) Can be used. Microfluidic devices may include microstructures formed using such molds.

The following examples describe certain features of various preferred embodiments of the present invention, and are not limiting in any sense. Rather, the scope of the invention is defined by the appended claims. Other materials and configurations not specifically disclosed herein are also contemplated. Such other embodiments will be apparent to those of ordinary skill in the art in view of the present disclosure.

Example 1 A three-dimensional microfluidic device was constructed using a stencil double-sided adhesive tape. In particular, a stencil was prepared by cutting channels from # 444 clear polyester film double sided tape (3M, StPaul, MN). A stencil of this tape was mounted on a 1/16 "thick acrylic sheet having a 0.04" diameter inlet hole positioned at one end of the stencil channel. A 2 mil thick Mylar® sheet was then laminated onto the stencil of the tape. The Mylar® substrate has a 0.04 ″ diameter hole positioned with the other end of the stencil channel. Other tape stencils that make up the channel are positioned on the Mylar® substrate. Finally, an acrylic cover substrate was placed on top of the tape, the acrylic resin substrate at the 0.04 "diameter outlet at one end of the stencil channel opposite the Mylar® hole. Has holes. FIG. 11 shows a micrograph of the microfluidic device in which water was passed. 11A or 11B show water on the first layer or the upper channel layer, respectively.

Example 2 A three-dimensional microfluidic device was constructed as follows. Modular components are laminated sheet tape (Avery Denni) with adhesive using a computer controlled plotter modified to have a cutting blade.
Son, LS10P, 73603) was excised to construct a channel and prepare a stencil. The seven types of these modules are designed to be modified (by simply reorienting them) to form a variety of microfluidic devices. As an example of this, two different microfluidic devices were constructed using these modules. In both cases, the first
The stencil was placed on a 1/16 "thick polycarbonate sheet substrate with 33 mil holes drilled as inlet holes. In one device, the remaining stencil was shown in sequence in Figure 12A (eg, , 1, 2, 3, 4, 4,
4,5,3,6,7,5,4,3,6,3,5,7,1), stacked so that the fluid is contiguous from one layer in place designed in a rotational shape I passed through the layers. The final substrate is an Avery Dennison LS10 with exit holes.
It was P tape. In this 17-layer microfluidic device, fluids enter and exit from the same direction. 12B and 12C are photomicrographs of the device at two stages of operation in which colored acetonitrile was passed through. FIG.
Using five similar modules, as shown in A, but changing their stacking order and orientation to be (1,2,3,4,4,4,4,7,1) Other devices were configured by. 13B and 13C are photomicrographs of the device at two stages of operation with colored water passed through.

Example 3 A microfluidic device consisting of a filter chamber was constructed. A stencil was prepared by cutting channels from a 1 mil thick vinyl sheet with adhesive on one side. The stencil was placed on a 1/16 "acrylic resin sheet with inlet holes positioned at either end of the filter.
The silica gel slurry was silica gel (JT Baker Chemical Co., P.) having an average particle diameter of 10 and 50 mM Nacl (aq) of 40 μm.
made by mixing hillsburg NJ) in the ratio of 1.
The filter chamber was filled with slurry by screening it in place. A similarly formed stencil was prepared by cutting a # 444 double-sided tape (3M). The tape stencil was typically positioned on and placed on a vinyl stencil, and an acrylic cover substrate was placed on the tape stencil. The colored fluid passed through the device (or filter chamber) but no leakage was observed.

Example 4 A microfluidic device consisting of a filter chamber was constructed. A stencil was prepared by cutting channels from a 1 mil thick vinyl sheet with adhesive on one side. The stencil was placed on a 1/16 "acrylic resin sheet with inlet and outlet holes located at either end of the filter chamber. Filter paper (Whatman Product # 107).
0) (Whatman Limited, England) sheet was cut and placed in the filter chamber area on the stencil. Two outlet holes provided with a built-in valve mechanism as described above were located on the opposite side of the filter. A Porex filter with a pore size of 25 μm was used as the valve mechanism. The sample passed through the filter. Remaining sample or multiple wash cycles (wa
sh cycle) was passed through the filter and then through the interior of the front chamber located below the filter chamber. When sufficient wash cycles have been applied, a plug of elution buffer is passed through the filter. The valve unit is then activated and the elution plug is passed inside the prechamber for analysis or further manipulation. This device is useful for purifying or concentrating nucleic acids, especially when the filter material is hydroxyapatite. FIG. 14 provides three photomicrographs (AC) of the device at various stages of operation.

Example 5 A microfluidic device was constructed as follows. Stencil has a thickness of 3
Heat-sealable with mill and annealed at 108 ° C
Nylon Tape (Product Number # 4220; Bemis Associates, I
nc. , Shirley, MA). The stencil was sandwiched between a 2 mil thick Mylar® substrate and a 1/16 ″ thick polycarbonate substrate with two inlet holes. An arbor press heated aluminum plate was attached. . The plate is
It was previously heated to 108 ° C. The device was positioned by pressing or pressing at a predetermined temperature for 2-3 seconds to seal the device. FIG. 15 is a photomicrograph of the assembled device with the fluid passing through.

Example 6 Referring to FIG. 7, shown is one designed for a biochemical (eg, protein) purification application. Two holes 100 and 101 (40 mil diameter each), either inlet or outlet holes, are drilled into a 1/8 "thick polycarbonate substrate 102. The stencil is modified to have a cutting blade. A computer-controlled plotter was used to excise the channels from a laminated sheet tape with adhesive (Avery Dennison, LS10P, 73603) (see FIG. 12 or FIG. 13). Filter chamber 104 is filled with Bio-Gel (R) HTP hydroxyapatite (Biorad, Hercules, CA). 80 mil diameter holes 106 are drilled into 1/8 "thick polycarbonate substrate 108, and ,
A sheet of PTFE (25 micron pore size) commercially available from Porex Technologies (Fairburn, GA) is used as the bottom substrate 110.

A 100 nl volume of protein solution in 10 mM phosphate buffer is added to the inlet hole 100. The protein binds to the hydroxyapatite filter 104, while other biomass does not. The volumes of chamber 105 and 80 mil hole 106 were sample volumes (10 nl) with 4x detergent (400 nl) containing buffer.
0nl). When this total volume of fluid hashes the filter 104, then 100 nl of concentrated salt buffer is injected. The molarity of salt in the elution buffer is different for different types of proteins, especially 400m
M phosphate buffer is sufficient. This solution elutes proteins from the filter 104. Then, the eluent is collected at the outlet hole 101. The bottom substrate 110 acts as a passive capillary valve by pumping the eluent through the outlet holes 101 by a capillary force.

Example 7 A three-dimensional microfluidic device was constructed that consisted of channels formed using both an electrical circuit board and an adhesive tape stencil. Referring to FIG. 16A, on the left is an exploded perspective view of each component of the device with channels 150 and 151 formed on an electronic circuit board 160 and covered with a silicone sealant coating. The inlet hole 200 and the outlet hole 201 form the channel 15 in the electronic circuit board.
It was formed inside 0 and 151, respectively. Acrylic cover plate substrate 202 with two holes 204 and 205 is covered with an electronic circuit board channel 150.
And mounted on 151. A generally horseshoe shaped channel 206 was constructed in a stencil 208 cut from a # 444 double sided tape (3M), which was positioned and adhered to the acrylic resin substrate 202. Finally, the acrylic resin substrate 210 was placed on the surface opposite the tape stencil 208. The assembled microfluidic device is shown to the right of FIG. 16A.

Fluid is injected into the inlet hole 200 and moves down the first electronic circuit board channel 150. Fluid travels through holes 204 and tape stencil channel 206
Pass through. Finally, the fluid flows through the holes 205 to the second electronic circuit board channel 1
It flows into the inside of 51 and flows out from the outlet hole 201. FIG. 16B is a micrograph of the device according to FIG. 16A with the fluid passing through.

Such devices can be used to achieve electrophoretic or electrokinetic separations. For example, electrodes may be provided at the inlet or outlet holes to apply the appropriate voltage. Because the optically transparent tape stencil layer 208 extends beyond the optically non-transmissive electronic circuit board, analysis of the fluid contained within the stencil channel 206 uses a variety of optical methods. It becomes possible by doing.

Example 8 A sample-splitting microfluidic device consisting of fork-shaped channels was constructed. A stencil as shown in FIG. 8B has a laminated sheet tape (Avery Dennison, LS10P, 7360) with adhesive using a computer controlled plotter modified to have a cutting blade.
It was constructed by cutting the outer peripheral region from 3). The device channels have a width of 25 mils. The stencil has the adhesive surface facing downwards and the inlet hole of the splitting device (Fig. 8).
1/16 "thick, with 33 mil holes positioned at
Of the acrylic resin substrate. Four 55 mil exit holes were cut using the same machine in a second piece of adhesive laminating tape, said tape with the exit holes approximately positioned at one end of the fork shaped channel. Placed on a split device. The fluid sample injected inside the device was divided into four approximately equal parts. 17A and 17B are photomicrographs showing water flowing through a splitting device as described above at two stages of operation.

Example 9 A microfluidic device capable of filtering a sample with a built-in valve was constructed. Laminated sheet tape with adhesive (Avery) using a computer controlled plotter where the stencil was modified to have a cutting blade.
Dennison, LS10P, 73603). Filter chamber 104 includes input channel 103 and output channel 10.
7 (see FIG. 7) in the stencil. Beyond the filter chamber 104, the output channel 107 is the chamber 105 and the channel 10.
Branched to 9. The stencil has channels 103 and 10 with the adhesive side facing down.
9 was placed on a 1/16 ″ thick acrylic substrate with 33 mil diameter holes positioned with one end (each hole 100 or 101 shown in FIG. 7). Filter chamber 104 was used in the previous embodiment. Filled as in 6. Once dried, a solid piece of LS10P was used to cover the filter chamber 104. Colored water was injected inside the device and the fluid flowed through the filter chamber 104 (Fig. 18A and 18B) and into chamber 105, selectively filling chamber 105 (see FIG. 18C). Passed inside channel 109 (see Figure 18D), because holes 100 and 101 in acrylic substrate 102 are located above the tape stencil, / Tape intermediate plane (aperture / tape interface) capillary force at the location were obviously stronger than the force in the channel 109. Devices in this regard are useful for purification or concentration of the sample.

Example 10 A microfluidic device was constructed using a tape stencil as part of the device and as a mold for silicon replication. Referring to FIG. 19A,
A silicon main mold was constructed using a vinyl sign cutter to make a stencil by cutting a dumbbell shaped portion having channels 250 and holes 251 from a 2 mil thick vinyl sheet. . The stencil was placed on an acrylic resin substrate. A small acrylic box was constructed around the stencil to keep the silicon from flowing from the surface. RTV615A silicone rubber (90%), RTV615B curing agent (10%) (General
Electric Comp. , Waterford, NY). The silicon was degassed, poured inside the box and allowed to cure overnight. FIG. 19B
The stencil 252 shown in Figure 6 was constructed using a technique similar to that described in Example 7. The final device is a silicon replica (repl) on a 1/16 "thick acrylic substrate with holes drilled in the silicon wells.
icate) was placed. A tape stencil 252 was mounted on the opposite side of the acrylic resin substrate with the adhesive side facing down. Inlet and outlet holes were punched in separate acrylic resin blocks and double-sided tape strips, which were mounted at opposite ends of the tape stencil. Fluid was injected through the device, but no leak was observed. FIG. 19C is a photomicrograph showing the fluid passing through the silicon replica. This mold manufacturing method is very useful because it is possible to form hundreds of molds on one vinyl sheet within a few minutes.

The invention described and claimed herein is not to be limited in scope by the particular embodiments disclosed herein. These embodiments are merely illustrative of some of the features of the present invention. All equivalent embodiments
Included within the scope of the invention. Indeed, the foregoing description will enable those skilled in the art to make various modifications of the invention in addition to those shown and described herein. Also, such modifications are included within the scope of the claims.

The disclosures of all referenced examples cited herein are incorporated by reference in their entirety.

[Brief description of drawings]

FIG. 1A shows the configuration of a microfluidic device consisting of a stencil sandwiched between two substrates. FIG. 1A is a perspective view showing the respective components of the device.

FIG. 1B shows a microfluidic device configuration consisting of a stencil sandwiched between two substrates. FIG. 1B is a three-dimensional perspective view showing the assembled state of the device of FIG. 1A.

FIG. 2 provides cross-sectional views (AD) showing a stencil coated with a sealant coating. In addition, adjacent substrates (cover plates) may be selectively covered as shown in FIG. 2D.

FIG. 2B provides cross-sectional views (AD) showing a stencil coated with a sealant coating. In addition, adjacent substrates (cover plates) may be selectively covered as shown in FIG. 2D.

FIG. 2C provides cross-sectional views (AD) showing a stencil coated with a sealant coating. In addition, adjacent substrates (cover plates) may be selectively covered as shown in FIG. 2D.

FIG. 2D provides cross-sectional views (AD) showing a stencil coated with a sealant coating. In addition, adjacent substrates (cover plates) may be selectively covered as shown in FIG. 2D.

FIG. 3 is a microfluidic where a first sealant coating material is used to coat the stencil or underlying substrate and a second sealant coating material is used to help seal the substrates together. It is sectional drawing which showed the device.

FIG. 4A shows a configuration of a three-dimensional microfluidic device including a stencil. FIG. 4A is an exploded perspective view showing each component.

FIG. 4B shows a configuration of a three-dimensional microfluidic device including a stencil. FIG. 4B is a three-dimensional perspective view showing the assembled state of the device of FIG. 4A.

FIG. 5A shows the construction of another three-dimensional microfluidic device consisting of a stencil. FIG. 5A is an exploded perspective view showing each component.

FIG. 5B shows the configuration of another three-dimensional microfluidic device consisting of a stencil. FIG. 5B is a three-dimensional perspective view showing the assembled state of the device of FIG. 5A.

FIG. 6A illustrates the use of a silk screen method to fill or coat an area (eg, filter chamber) of a microfluidic device.
FIG. 6A shows each component.

FIG. 6B illustrates the use of the silk screen method to fill or coat a predetermined area (eg, filter chamber) of a microfluidic device.
FIG. 6B shows a positioning process for silkscreening a panel of devices.

FIG. 7 is an exploded perspective view of a microfluidic device having an integral (ie, “built-in”) valve mechanism.

FIG. 8 is a top view of a microfluidic device that includes a fork-shaped channel that can divide a sample into four approximately equal portions.

FIG. 9 illustrates components of a microfluidic device having an integral (ie, “built-in”) valve mechanism.

FIG. 10 is an exploded view showing an alignment method using a peg-hole alignment method to ensure proper alignment of various layers and components of a microfluidic device.

FIG. 11A is a micrograph of a microfluidic device including a sandwiched stencil at some stage of fluid passing manipulation.

FIG. 11B is a photomicrograph of a microfluidic device containing a sandwiched stencil at some stage of fluid passing manipulation.

FIG. 12A shows the construction of a microfluidic device constructed using 18 sheets of stencil.

FIG. 12B is a photomicrograph of the device of FIG. 12A at a stage in the operation of passing acetonitrile through the device.

FIG. 12C is a photomicrograph of the device of FIG. 12A at a stage in the process of passing acetonitrile through the device.

FIG. 13A shows a configuration of a microfluidic device constructed using 9 stencils.

13B is a photomicrograph of the device of FIG. 13A at some stage of operation with water passing through.

FIG. 13C is a photomicrograph of the device of FIG. 13A at some stage of operation with water passing through.

14A-C are photomicrographs showing water passing through a microfluidic device containing a chamber filled with silica gel.

FIG. 15 is a photomicrograph of a microfluidic device constructed using a stencil containing a thermal (eg, heat activated) tape.

FIG. 16A shows a microfluidic device configuration consisting of both an electronic circuit board and a stencil. FIG. 16A shows an exploded perspective view on the left side and an assembled perspective view on the right side.

FIG. 16B shows a microfluidic device configuration consisting of both an electronic circuit board and a stencil. FIG. 16B shows a state in which the fluid is passing through the inside of FIG.
6A is a micrograph of the device shown in FIG. 6A.

FIG. 17A is a micrograph of a microfluidic device containing fork-shaped channels capable of disrupting or splitting a fluid sample at one stage of operation.

FIG. 17B is a micrograph of a microfluidic device containing fork-shaped channels capable of disrupting or splitting a fluid sample at one stage of operation.

FIG. 18A is a micrograph of a microfluidic device having a built-in filter or valve with fluid passing through at some stage of operation.

FIG. 18B is a micrograph of a microfluidic device having a built-in filter or valve with fluid passing through at some stage of operation.

FIG. 18C is a micrograph of a microfluidic device having a built-in filter or valve with fluid passing through at some stage of operation.

FIG. 18D is a micrograph of a microfluidic device having a built-in filter or valve with fluid passing through at some stage of operation.

FIG. 19A shows the use of a stencil as a mold to create a microfluidic replica.

FIG. 19B shows the use of a stencil as a mold to create a microfluidic replica.

FIG. 19C is a micrograph of a silicon microfluidic replica made using the mold with fluid passing through.

─────────────────────────────────────────────────── ─── Continued front page    (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, I T, LU, MC, NL, PT, SE), OA (BF, BJ , CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, K E, LS, MW, MZ, SD, SL, SZ, TZ, UG , ZW), EA (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, BZ, C A, CH, CN, CR, CU, CZ, DE, DK, DM , DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, K E, KG, KP, KR, KZ, LC, LK, LR, LS , LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, RO, R U, SD, SE, SG, SI, SK, SL, TJ, TM , TR, TT, TZ, UA, UG, UZ, VN, YU, ZA, ZW (72) Inventor Mercy Petzut             United States 91106 Pa, California             Sadina, number 103, South Katari             Na 442 F-term (reference) 2G058 DA07 DA09 GA12                 2G060 AA05 AC10 AE40 AF06 AF10                       FA01 KA05

Claims (56)

[Claims] 1. A stencil comprising: first and second substrates; and at least one stencil disposed between the first and second substrates to form one or more sealed microstructures. ,
A microfluidic device adhered to at least one of the first and second substrates with an adhesive. 2. The microfluidic device of claim 1, comprising a plurality of stencils. 3. The microfluidic device according to claim 1, wherein the first and second substrates are substantially flat. 4. The adhesive according to claim 1, wherein the adhesive is an adhesive selected from the group consisting of a rubber-based adhesive, an acrylic resin-based adhesive, and a resin-based adhesive. Microfluidic device. 5. The microfluidic device according to claim 1, wherein the stencil has a self-adhesive property. 6. The microfluidic device according to claim 1, wherein the stencil comprises an adhesive tape. 7. The microfluidic device according to claim 6, wherein the adhesive tape has an adhesive on one surface. 8. The microfluidic device according to claim 6, wherein the adhesive tape has an adhesive on both surfaces. 9. The microfluidic device of claim 6, wherein the adhesive tape is selected from the group consisting of a pressure sensitive tape, a heat activated tape, a chemically activated tape and a photo activated tape. 10. The microfluidic device according to claim 6, wherein the adhesive tape is made of a material selected from the group consisting of Mylar, nylon, and polyester. 11. The microfluidic device according to claim 1, wherein the stencil and at least one of the first and second substrates are ultrasonically welded to each other. 12. The microfluidic device of claim 1, wherein the stencil is made of a material selected from the group consisting of polymer, paper, fabric and foil foil. 13. The stencil includes mylar, polyester, polyimide, vinyl, acrylic resin, polycarbonate, Teflon, Kapton, polyurethane, polyethylene, polypropylene, polyvinylidene fluoride, polytetrafluoroethylene, nylon, polyether sulfone, and acetal. Microfluidic according to claim 12, characterized in that it consists of a polymer selected from the group consisting of polymeric polyesterimides, polysulfones, polyphenylsulfones, ABS, polyvinylidene fluorides, polyphenylene oxides and derivatives thereof. device. 14. The microfluidic device of claim 12, wherein the stencil comprises a fluorinated polymer. 15. The microfluidic device according to claim 1, wherein the stencil has an elastomeric property. 16. The microfluidic device of claim 15, wherein the stencil comprises an elastomeric material selected from the group consisting of rubber, viton, and silicone. 17. The microfluidic device of claim 1, wherein the stencil is die cut. 18. At least one of the first and second substrates is made of a material selected from the group consisting of Mylar, FR-4, polyester, glass, acrylic resin, polycarbonate, and fiber glass. The microfluidic device according to claim 1. 19. The microfluidic device of claim 1, further comprising a sealant coating on at least a portion of one or more of the stencil, the first substrate and the second substrate. 20. The microfluidic device of claim 19, wherein the sealant coating comprises a silicone material. 21. The sealant coating is Teflon, abatrel, liquin, fluorocarbon, fluorothermoplastic, polyvinylidene fluoride, acrylic resin, wax, epoxy resin, solder, polymer, paint, oil, varnish. 20. The microfluidic device of claim 19, comprising one or more materials selected from the group consisting of: 22. The microfluidic device of claim 19, wherein the sealant coating is formed by spin film deposition. 23. The microfluidic device of claim 19, wherein the sealant coating is formed by spraying. 24. The microfluidic device of claim 19, wherein the sealant coating is formed by dipping. 25. The microfluidic device of claim 1, wherein the microstructure comprises channels or chambers. 26. The microfluidic device of claim 1, wherein the microstructure is at least partially filled with a filling material. 27. The microfluidic device of claim 26, wherein the filling material is a filter material. 28. The filter material is polycarbonate, acrylic resin,
The microfluidic device according to claim 27, wherein the microfluidic device is selected from the group consisting of acrylamide, polyurethane, polyethylene, polypropylene, polyvinylidene fluoride, polytetrafluoroethylene, Nahion, nylon, and polyethersulfone. 29. The microfluidic device of claim 27, wherein the filter material is selected from the group consisting of agarose, alginate, starch and carrageenan. 30. The microfluidic device of claim 27, wherein the filter material is silica gel. 31. The microfluidic device of claim 27, wherein the filter material is selected from the group consisting of Sephadex or Sephakill. 32. The microfluidic device of claim 27, wherein the filter material is hydroxyapatite. 33. The microfluidic device according to claim 26, wherein the filling material is formed by a silk screen. 34. The microfluidic device of claim 26, wherein the fill material is formed by lithography. 35. The microfluidic device according to claim 26, wherein the filling material is formed by a pick and place method. 36. The microfluidic device of claim 1, wherein the at least one substrate has one or more holes. 37. The microfluidic device of claim 1, further comprising one or more valves. 38. The microfluidic device of claim 1, wherein the microstructure comprises fork-shaped channels. 39. The microfluidic device of claim 1, further comprising at least one electrode. 40. The microfluidic device according to claim 39, wherein the electrode is for detecting or measuring an electrical property of a fluid. 41. The microfluidic device of claim 39, wherein the electrodes are for promoting electrophoretic or electrokinetic flow of fluid. 42. The portion of at least one of the first substrate and the second substrate allows passage of an optical signal.
The microfluidic device according to. 43. The microfluidic device of claim 1, further comprising one or more additional substrates sealingly engaged. 44. The microfluidic device of claim 43, wherein the substrate comprises an electronic circuit board having microstructures on its surface. 45. The modular microfluidic device of claim 1, connectable to another microfluidic device. 46. A plurality of microfluidic devices according to claim 1, wherein at least two microfluidic devices of the microfluidic devices are formed such that fluid can move between them. And a microfluidic system. 47. Two or more microfluidic devices are stacked 3
47. The microfluidic system of claim 46, forming a dimensional microfluidic system. 48. A manufacturing method for simultaneously manufacturing a plurality of microfluidic devices, wherein a first substrate is provided, and one or more panels made of a stencil array are laminated on the first substrate, A method of manufacturing, comprising laminating a second substrate on a plurality of panels to form a plurality of microstructures therebetween. 49. At least one of the first and second substrates is 1
49. The manufacturing method according to claim 48, which has one or a plurality of holes. 50. At least one of the panels is positioned on at least one of the first and second substrates to allow fluid movement between the holes and the microstructures. Item 49. The manufacturing method according to Item 49. 51. The manufacturing method of claim 50, wherein the positioning is provided by peg-hole alignment. 52. The manufacturing method according to claim 48, wherein the stacking is performed by a converter. 53. A microfluidic device prepared by the manufacturing method according to claim 48. 54. A mold provided by using at least a part of the stencil according to claim 1 as a mold for forming a mold. 55. The mold according to claim 54, which is made of a silicon material. 56. A microfluidic device comprising a microstructure provided using the mold of claim 54.