JP5585204B2 - Microchip and manufacturing method thereof - Google Patents

Description

The present invention relates to a microchip and a manufacturing method thereof. In particular, the present invention relates to a microchip having a microchannel formed inside a substrate and a method for manufacturing the microchip.

Microchips called micro-TAS (Micro Total Analysis System), Lab-on-a-chip, and MEMS (Micro Electro Mechanical Systems), in which semiconductor manufacturing technology is applied to channel formation, have been studied and put into practical use.

These microchips have a structure having a micrometer-order width channel called a microchannel inside a chip having a size of several cm square, and the microchannels are joined or branched. A microchip is a system that allows a small amount of solution to flow through a microchannel to perform reaction, separation, and analysis of the solution. Therefore, the microchip is very useful because it can reduce the amount of sample and increase the efficiency of the reaction. In addition, since the entire device including the microchip can be downsized by reducing the size of the flow path, application to various applications is possible.

Silicon, glass, resin, or the like is used as a material for the microchip. Resin microchips have the advantage of being easy to mass-produce, but have low heat resistance, swelling and corrosion caused by organic solvents, and dissolution of adhesive components used for bonding. There is a problem that deformation of the microchip occurs due to pressure or external pressure.

Since the silicon microchip is made of an inorganic material, it has high heat resistance, does not swell or corrode with an organic solvent, and hardly deforms due to fluid pressure or external pressure. Such a silicon microchip can be manufactured using the MEMS technology, and is excellent in productivity.

On the other hand, a glass microchip can provide a microchip that has high heat resistance, does not swell or corrode with an organic solvent, or hardly deforms due to fluid pressure or external pressure, and is transparent. Since it is a material, optical observation in the flow path is also possible. By using a glass microchip, for example, it can be used for imaging of cells, proteins, DNA, etc. using a fluorescent substance or the like.

Patent Document 1 discloses that a microchip capable of such optical observation is manufactured by combining a glass substrate and a silicon-containing substrate. Patent Document 2 discloses that a flow path having a complicated shape is formed by combining a glass substrate and a silicon substrate. A microchip combining such a glass substrate and a silicon substrate has higher heat resistance than a resin microchip, does not swell or corrode due to organic solvents, or hardly deforms due to fluid pressure or external pressure. Is excellent.

However, in these conventional microchips, a flow path is formed on the surface of a base material made of silicon, glass, resin, etc., and at least one more base material needs to be bonded to cover this flow path. .

JP 2010-25688 A JP 2010-29780 A

As described above, the conventional microchip has a large number of base materials necessary for forming one microchip, and as a result, it becomes thick and becomes an obstacle to miniaturization. Moreover, in the microchip which combined the glass base material and the silicon base material, when joining base materials directly, it is necessary to use the special glass base material containing a mobile ion, and a joining process is needed. Further, the manufacturing cost and the necessary equipment for processing the silicon base material forming the flow path increase, resulting in an increase in manufacturing cost.

Therefore, the present invention has been made in view of the above problems, and it is an object of the present invention to provide a microchip that has high heat resistance and chemical resistance and can be manufactured with a small number of manufacturing steps, and a manufacturing method thereof.

According to one embodiment of the present invention, at least two on the surface or side of the substrate inside a single transparent substrate selected from borosilicate glass, soda lime, alumina borosilicate glass, quartz glass and sapphire . have a opening, have a flow path that is formed to bend in the horizontal direction and / or direction perpendicular to the upper surface of the substrate in the substrate, formed from a surface of the flow passage protrudes integrally a substrate microchip, characterized in that the perforated the flow path plate-like or columnar structure is are provided.

Thereby, the microchip which formed the flow path in the inside of a single transparent base material is provided, without bonding and lamination | stacking of a base material. Therefore, as in the case of directly joining the substrates, there is no problem such as liquid leakage due to poor bonding due to voids at the bonding interface. Further, as in the case of anodically bonding a glass substrate and a silicon substrate, alkali precipitation occurs in a portion corresponding to the opening, and there is no problem depending on the properties of the test object. Further, unlike the case where the base material is bonded using an adhesive, the flow path characteristics are not deteriorated by the adhesive, and the observation of the test object in the flow path is not hindered by the adhesive. Further, the microchip can be miniaturized by meandering in one base material to form a flow path, and since it is a single base material, it is harder to break than an elongated capillary and is easy to handle. Is provided. Furthermore, by forming the structure in the flow path, for example, the flow rate and pressure of the fluid can be controlled.

Also, according to one embodiment of the present invention, a single transparent selected from borosilicate glass, soda lime, alumina borosilicate glass, quartz glass and sapphire isotropically etched after being modified by laser. It is formed inside the substrate, and have at least two openings in the surface or side of the substrate, is formed to bend in the horizontal direction and / or direction perpendicular to the upper surface of the substrate in the substrate have a flow channel, microchip from the surface of the flow passage protrudes characterized in that have a base plate and formed integrally with or columnar structure in the flow path is provided.

A conventional microchip in which a glass substrate and a silicon substrate are combined has a large number of substrates to be used, so that the manufacturing process is complicated. However, according to an embodiment of the present invention, a single substrate is formed with a small number of manufacturing processes. A microchip having a flow path formed therein is provided. Further, the microchip can be miniaturized by meandering in one base material to form a flow path, and since it is a single base material, it is harder to break than an elongated capillary and is easy to handle. Is provided. Furthermore, by forming the structure in the flow path, for example, the flow rate and pressure of the fluid can be controlled.

The cross-sectional shape of the channel of the microchip may be a circle, an ellipse, a polygon, a ring, or a donut.

In the conventional semiconductor manufacturing technology, it is relatively easy to form a rectangular flow path by using anisotropic etching, but in order to form a circular flow path inside the substrate, a complicated manufacturing process is required. Necessary. According to an embodiment of the present invention, by using wet etching, which is isotropic etching, the cross section of the formed flow path has a substantially circular shape, and a microchip with a simple manufacturing process is provided. Further, in a polygonal cross-sectional flow path, a plurality of flow paths can be compactly integrated on a single substrate by using the geometrical arrangement of the flow paths. In a ring-shaped or donut-shaped cross-sectional flow path, fluids with different temperatures can be flowed in each flow path, and used for heat exchange between the fluids. it can.

The cross-sectional shape of the channel of the microchip may be different depending on the position of the substrate.

As a result, the degree of freedom in designing the microchip is increased, and the development of microchips for various applications can be made possible.

A plurality of structures may be provided in the flow path of the microchip, and the plurality of structures may have structures having different sizes.

As a result, the fluid can be agitated by forming a plurality of structures in the channel and making the sizes of the structures different. The stirring effect can be enhanced by forming a plurality of plate-like structures at a predetermined interval inside one flow path.

Further, according to one embodiment of the present invention, a laser beam is irradiated inside a single substrate selected from borosilicate glass, soda lime, alumina borosilicate glass, quartz glass, and sapphire, and a part of the substrate is formed. reforming, a portion of the substrate is removed by etching, to have at least two openings in the surface or side of the substrate, the horizontal and / or vertical direction with respect to the upper surface of the substrate in the substrate A channel formed so as to be bent is formed inside the substrate, protrudes from the surface of the channel, and a plate-like or columnar structure formed integrally with the substrate is formed in the channel. A method for manufacturing a microchip is provided.

Thereby, a flow path can be formed in the inside of a single transparent base material, without bonding and lamination | stacking of a base material. In addition, a microchip that has high heat resistance and chemical resistance and can be manufactured using an inexpensive material is provided. Further, the microchip can be downsized by meandering in one base material to form a flow path, and since it is a single base material, it is harder to break than an elongated capillary and is easy to handle. Can be manufactured. In order to form a structure in a flow path by a conventional semiconductor manufacturing technique, a complicated manufacturing process is required. According to one embodiment of the present invention, it is possible to form a complicated flow path more easily than in the past. Accordingly, the degree of freedom in designing a microchip having functionality in the flow path is increased, and it becomes possible to develop microchips for various applications.

In the microchip manufacturing method, the laser may be a pulse laser of 1000 femtoseconds or less.

In the femtosecond laser, nonlinearity becomes remarkable in the interaction between the laser and the substance, so that processing and thermal influence using multiphoton absorption can be ignored. The femtosecond laser can irradiate a laser beam to an extremely small region as compared with a carbon dioxide laser or a YAG laser, and the processing accuracy is remarkably high. By using a femtosecond laser, energy is instantaneously injected into the object to be processed, and it is not subject to thermal and chemical damage. “Modification” can be performed to change to a sparse state.

In the microchip manufacturing method, the cross-sectional shape of the channel may be formed in a circular shape, an elliptical shape, a polygonal shape, a ring shape, or a donut shape.

In order to form a flow path having a complicated cross-sectional shape by a conventional semiconductor manufacturing technique, a complicated manufacturing process is required. According to one embodiment of the present invention, it is possible to form a complicated flow path more easily than in the past. Therefore, the degree of freedom in the design of the microchip is increased, and it becomes possible to develop microchips for various uses.

In the microchip manufacturing method, the cross-sectional shape of the flow path may be formed to be different depending on the position of the substrate.

In order to form a combination of a plurality of channels having various cross-sectional shapes within one microchip, the conventional semiconductor manufacturing technology requires a complicated manufacturing process. According to one embodiment of the present invention, it is possible to form a complicated flow path more easily than in the past. Therefore, the degree of freedom in the design of the microchip is increased, and it becomes possible to develop microchips for various uses.

In the microchip manufacturing method, a plurality of structures having different sizes may be formed in the channel.

In order to form a structure in a flow path by a conventional semiconductor manufacturing technique, a complicated manufacturing process is required. According to one embodiment of the present invention, it is possible to form a complicated flow path more easily than in the past. Accordingly, the degree of freedom in designing a microchip having functionality in the flow path is increased, and it becomes possible to develop microchips for various applications.

According to the present invention, it is possible to provide a microchip that has high heat resistance and chemical resistance and can be manufactured with a small number of manufacturing steps, and a manufacturing method thereof.

It is the schematic diagram of the microchip 100 of this invention which concerns on one Embodiment, (a) is the figure which looked at the microchip 100 from diagonally upward direction, (b) is the cut surface in AA ', (c ) Is a cut surface at BB ′. It is a figure which shows irradiation of the pulse laser of the manufacturing process of the microchip 100 of this invention which concerns on one Embodiment, (a) is the figure which looked at the microchip 100 from diagonally upward direction, (b) is AA '. It is a cut surface. It is a figure which shows the relationship between the irradiation method of a laser beam, and the shape of the modification part formed, (a) shows the figure which scans the focus of a laser beam so that it may not propagate through the modification part formed previously. (B) shows the figure which scans the focus of a laser beam so that it may propagate through the modification part formed previously. The cross-sectional shape of the flow path 21 of this invention which concerns on one Embodiment is shown, (a) shows the flow path 21 of various cross-sectional shapes equivalent to the cut surface in AA 'of Fig.1 (a), (b ) Shows a flow path 23 having a groove-shaped cross-sectional shape. The cross-sectional shape of the flow path 31 of this invention which concerns on one Embodiment is shown, (a) is sectional drawing of the flow path 31 which flows a fluid in a ring shape, (b) is sectional drawing of the double flow path 33. is there. It is a figure which shows arrangement | positioning of the flow path of this invention which concerns on one Embodiment, (a) is sectional drawing which shows a mode that the several flow path 41 was formed in the inside of the base material 1, (b) is a base material 1 is a cross-sectional view showing a state in which a plurality of flow paths 43 are formed inside 1. FIG. An example of a flow path in which a structure is formed inside the present invention according to an embodiment is shown, (a) shows a flow path 51 in which a stepped structure 81 is formed inside one flow path, and (b) ) Shows a flow channel 53 in which a plate-like structure 83 is formed inside one flow channel. 1 shows a microchip 200 of the present invention according to an embodiment, wherein (a) is a view of the microchip 200 as viewed from above, (b) is a cut surface at AA ′, and (c) is BB ′. It is a cut surface at. 4A and 4B are schematic views of a microchip 300 according to an embodiment of the present invention, where FIG. 5A is a top view and FIG. 5B is a cross-sectional view taken along line BB ′ of FIG. It is a schematic diagram of the microchip 400 of this invention which concerns on one Example, (a) is a top view, (b) shows sectional drawing in BB 'of (a). (A)-(c) is a figure which shows the observation method of the chromosomal DNA using the microchip 400. FIG. It is a schematic diagram of the microchip 500 of this invention which concerns on one Example, (a) is a top view, (b) shows sectional drawing in BB 'of (a).

Hereinafter, a microchip and a manufacturing method thereof according to the present invention will be described with reference to the drawings. However, the microchip of the present invention can be implemented in many different modes and should not be construed as being limited to the description of the embodiments and examples shown below. Note that in the drawings referred to in this embodiment mode and examples, the same portions or portions having similar functions are denoted by the same reference numerals, and repetitive description thereof is omitted.

As a result of intensive studies on the above-described problems, a method of forming a flow path inside a single substrate has been used. That is, since the above-mentioned microchip combining a glass substrate and a silicon substrate has a large number of substrates, the manufacturing process is complicated, but if a flow path can be formed inside a single substrate with a small number of manufacturing processes. The problem can be solved. It has been found that such a manufacturing method can be realized by a combination of pulsed laser irradiation and wet etching.

(Embodiment 1)
FIG. 1 is a schematic diagram of a microchip 100 according to this embodiment of the present invention. FIG. 1A is a view of the microchip 100 viewed obliquely from above, FIG. 1B is a cut surface at AA ′, and FIG. 1C is a cut surface at BB ′. For example, the microchip 100 has a meandering channel 11 inside a single substrate 1, and the channel 11 is connected to an opening 90 formed on the upper surface of the substrate 1.

The channel 11 is a micro channel, and is a channel for flowing a fluid sample. By disposing the channel 11 so as to meander inside the substrate 1, a channel longer than the length of the substrate 1 in the BB ′ direction can be formed. When the length of the flow path 11 may be short, it may be arranged so as to connect the two openings 90 in a straight line. Further, the flow path 11 may be formed so as to have an inclination that decreases from B to the B ′ direction, for example, according to the viscosity of the sample.

The opening 90 can be used, for example, to add a sample to be passed through the flow path 11. The opening 90 may be formed perpendicular to the upper surface of the substrate 1 or may be formed so as to be inclined according to the viscosity of the sample. The opening 90 may be formed on the side surface of the microchip 100.

The microchip 100 according to this embodiment of the present invention can be manufactured by combining pulsed laser irradiation and wet etching. FIG. 2 is a diagram showing pulsed laser irradiation in the manufacturing process of the microchip 100 according to this embodiment of the present invention. FIG. 2A is a view of the microchip 100 viewed obliquely from above, and FIG. 2B is a cut surface taken along AA ′.

Here, the substrate 1 of the microchip 100 is a transparent substrate having high chemical resistance, that is, glass having high acid resistance and alkali resistance, such as borosilicate glass, soda lime, and alumina borosilicate glass. Quartz glass or the like can be used. Moreover, single crystal base materials, such as sapphire, may be sufficient. Since these base materials have translucency, the inside of the base material can be irradiated with laser light.

In order to form the flow path 11 in the microchip 100, a laser beam 5 is irradiated to a predetermined position inside the substrate 1 as shown in FIGS. 1 (a) and 1 (b). As the laser beam 5 according to the present embodiment, a pulse laser (femtosecond laser) having a pulse width of 1000 femtoseconds or less can be used. In the femtosecond laser, nonlinearity becomes remarkable in the interaction between the laser and the substance, so that processing and thermal influence using multiphoton absorption can be ignored. The femtosecond laser can irradiate a laser beam to an extremely small region as compared with a carbon dioxide laser or a YAG laser, and the processing accuracy is remarkably high. By using a femtosecond laser, energy is instantaneously injected into the object to be processed, and it is not subject to thermal and chemical damage. “Modification” can be performed to change to a sparse state.

When the substrate 1 is irradiated with the laser beam 5, the substrate 1 at the position of the focal point 6 of the laser beam 5 is modified, and the modified portion 7 can be formed. The modified portion 7 corresponding to the flow path 11 can be formed by scanning the laser beam 5 at a desired position inside the substrate 1. The modified portion 7 has a property that it is more easily etched than the unmodified portion of the substrate 1. Scanning with the laser beam 5 can be performed in the X-axis direction, the Y-axis direction, and the Z-axis direction, whereby the desired modified portion 7 can be formed. By scanning the laser beam 5 with high accuracy, it is possible to form the modified portion 7 for forming the flow path 11 with less unevenness on the inner surface.

Next, the base material 1 on which the modified portion 7 is formed is etched. The etching according to the present embodiment is preferably wet etching which is isotropic etching. As the etchant, hydrofluoric acid or a hydrofluoric acid-based solution can be used. The modified portion 7 is etched faster than the unmodified portion, and a flow path 11 having a width on the order of micrometers is formed on the substrate 1. By increasing the etching time, unevenness on the surface inside the flow path 11 can be reduced. Moreover, in order to prevent the etching of the surface of the base material 1 by an etching liquid, you may form a protective film on the surface of the base material 1 in which the modification part 7 was formed before an etching process. Etching liquid infiltrates into the inside of the base material 1 from the portion (corresponding to the opening 90) on the surface of the base material 1 where the protective film is not formed, and the flow path 11 can be formed by etching the modified portion 7. . The protective film can be formed of a material having resistance to an etching solution, and for example, chromium, gold, or an alloy thereof can be used.

In the conventional semiconductor manufacturing technology, it is relatively easy to form a rectangular flow path by using anisotropic etching, but in order to form a circular flow path inside the substrate, a complicated manufacturing process is required. Necessary. The microchip 100 according to this embodiment of the present invention uses wet etching, which is isotropic etching, so that the cross-section of the formed flow path 11 has a substantially circular shape, and the manufacturing process is simple.

Since the microchip 100 includes the meandering flow path 11 inside the substrate 1, a long flow path can be accommodated in a compact manner, and can be used instead of a microcapillary, for example. For example, capillary electrophoresis, capillary DNA sequencer, and the like require the length of the capillary in order to obtain high separation performance, so that the apparatus becomes correspondingly large and becomes an obstacle to miniaturization. The microchip 100 can be downsized by meandering in a single base material to form a flow path, and since it is a single base material, it is harder to break than an elongated capillary and is easy to handle. It becomes.

The microchip 100 according to this embodiment of the present invention combines a pulse laser irradiation and wet etching to form a substantially circular shape inside a single substrate without bonding or stacking the substrates. There is an excellent effect that a meandering channel having a cross-sectional shape can be formed. In addition, according to the present embodiment of the present invention, a microchip that has high heat resistance and chemical resistance and can be manufactured using an inexpensive material is provided.

In the conventional microchip for joining a plurality of base materials, when the base materials are directly joined, if there is a void at the joint interface, a joint failure is likely to occur, and problems such as liquid leakage may occur. Further, when anodically bonding a glass substrate and a silicon substrate, alkali precipitation occurs in a portion corresponding to the opening 90, which may cause a problem depending on the properties of the test object. Furthermore, when joining a base material using an adhesive agent, the deterioration of the flow path characteristic by an adhesive agent or the observation of the test object in a flow path may be obstructed by an adhesive agent. On the other hand, since the microchip 100 according to the present embodiment of the present invention can form a flow path inside a single substrate without bonding and stacking the substrates, such a problem is Does not occur.

(Embodiment 2)
In this embodiment, an example of forming a flow path having a complicated shape will be described. FIG. 3 is a diagram showing the relationship between the laser beam irradiation method and the shape of the modified portion formed. FIG. 3A shows a view of scanning the focal point of the laser beam so as not to propagate the previously formed modified portion, and FIG. 3B shows a laser beam propagating through the previously formed modified portion. FIG. 6 shows a diagram of scanning the focus of light.

As shown in FIG. 3A, when scanning in one direction so that the focal point 6 of the laser beam 5 does not propagate through the modified portion 7 of the substrate 1, the refractive index of the substrate 1 is constant. The beam diameter of the laser beam 5 is not expanded, and the diameter of the modified portion 7 can be reduced. On the other hand, as shown in FIG. 3B, when the focal point 6 of the laser beam 5 is scanned inside the base material 1 so as to propagate through the modified part 7, the modified part 7 already formed is Since the refractive index is different from 1 and the laser beam 5 incident on the modified portion 7 is reflected and refracted, the beam diameter of the laser beam 5 is expanded and the diameter of the modified portion 7 is increased. Therefore, a larger modified portion 7 can be formed by scanning the focal point 6 of the laser beam 5 so as to propagate through the previously formed modified portion 7.

By etching the modified portion 7 formed in this way, a flow path having a complicated shape can be formed inside the substrate 1. Therefore, if the relationship between the irradiation method of the laser beam 5 and the shape of the modified portion 7 to be formed is utilized, various shapes of flow paths can be formed. An example is shown in FIG. FIG. 4A shows examples of the flow paths 21 having various cross-sectional shapes corresponding to the cut surface at AA ′ of FIG. By using the relationship between the irradiation method of the laser beam 5 and the shape of the modified portion 7 to be formed, it is possible to form the flow path 21 having various cross-sectional shapes. Here, since the wet etching is used to form the flow path 21 according to the present embodiment, the flow path 21 having a triangular, rectangular, or polygonal cross-sectional shape is compared with the flow path using anisotropic etching. And the corners are rounded.

In a circular or elliptical cross-sectional flow path, the pressure of the fluid flowing in the flow path is substantially uniformly applied to the entire wall surface of the flow path, so that the microchip can be used under high pressure conditions. On the other hand, with a polygonal cross-sectional flow path, a plurality of flow paths can be compactly integrated on a single substrate by using the geometrical arrangement of the flow paths described later. For example, by forming a honeycomb structure using a flow path having a regular hexagonal cross section in one substrate, the flow paths can be integrated without impairing the strength of the microchip.

FIG. 4B is an example of a channel having a more complicated cross-sectional shape, and shows a channel 23 having a groove-shaped cross-sectional shape. Although FIG. 4B illustrates the groove-shaped cross-sectional shape having a convex portion in the bottom surface direction of the base material 1, the groove-shaped cross-sectional flow channel 23 having the convex portion in the top surface direction of the base material 1 can also be formed. . Since the flow path 23 having such a groove-shaped cross-sectional shape has a large surface area, it is effective for increasing the contact area between the fluid and the flow path, for example.

FIG. 5 is a cross-sectional view of the flow path 31 through which the fluid flows in a ring shape or a donut shape, and FIG. 5A corresponds to a cut surface at AA ′ in FIG. The flow path 31 forms the ring-shaped modified portion 7 by scanning the focal point 6 of the laser beam 5 so as to leave a base material portion that is not modified at the center. By removing the ring-shaped modified portion 7 by etching, a flow path 31 having a ring-shaped cross-sectional shape can be formed. The flow channel 31 shown in FIG. 5A is an example, and can be formed in a polygonal ring shape. Further, the shape of the outer periphery may be different from the shape of the inner periphery, for example, the outer periphery of the flow path is circular and the inner periphery of the base material 1 is rectangular.

FIG. 5B is a cross-sectional view of the double flow path 33. The channel 33 has a channel 33b inside the channel 33a. The flow path 33 can be used, for example, to flow fluids having different temperatures through the flow paths 33a and 33b and to exchange heat between the fluids. Also, different types of liquids and fluids with different viscosities can be passed through the flow path 33a and the flow path 33b. The flow path 33 shown in FIG. 5B is an example, and can be formed in any number of structures as long as the thickness of the base material 1 allows. Moreover, the cross-sectional shape of the flow path 33a and the flow path 33b is not limited to a circle, and can be formed in a desired shape as shown in FIG.

In order to form a flow path having a complicated cross-sectional shape as described above by a conventional semiconductor manufacturing technique, a complicated manufacturing process is required. By using the flow path forming method of the present invention according to this embodiment, a complicated flow path can be formed more easily than in the past. Therefore, by using the flow path of the present invention according to the present embodiment, the degree of freedom in designing the microchip is increased, and an excellent effect that enables the development of microchips for various uses is achieved.

(Embodiment 3)
In the present embodiment, a method of arranging a plurality of flow paths having various cross-sectional shapes in one microchip by applying the complicated flow path forming method described in the second embodiment will be described.

FIG. 6 is a diagram showing the arrangement of the flow paths according to this embodiment of the present invention. FIG. 6A is a cross-sectional view showing a state in which a plurality of flow paths 41 are formed inside the substrate 1. FIG. 6A corresponds to a cut surface at AA ′ in FIG. The channel 41 is formed so that a plurality of channels are arranged in a direction perpendicular to the bottom surface of the substrate 1. As shown in FIG. 6 (a), a plurality of channels having the same cross-sectional shape can be formed, or a plurality of channels having different cross-sectional shapes can be formed.

FIG. 6B is a cross-sectional view showing a state in which a plurality of flow paths 43 are formed inside the base material 1. FIG. 6B corresponds to a cut surface at AA ′ in FIG. The flow path 43 shows an example in which a plurality of flow paths are arranged in a compact manner by geometrically arranging a plurality of flow paths. With such a geometrical arrangement of the flow path 43, it is possible to reduce the size of the microchip even if the total length of the flow path is increased. Further, by using the microchip in which the efficiently integrated flow paths 43 are formed, as a result, it is possible to promote downsizing of the apparatus using the microchip. Note that the flow path 43 shown in FIG. 5B is an example, and other geometric flow paths can be arranged.

In order to form a combination of a plurality of channels having various cross-sectional shapes inside one microchip as described above, a conventional semiconductor manufacturing technique requires a complicated manufacturing process. By applying the flow path forming method of the present invention according to the second embodiment to the flow path arrangement of the present embodiment, a complicated flow path can be formed more easily than in the past. Therefore, by using the flow path of the present invention according to the present embodiment, the degree of freedom in designing the microchip is increased, and an excellent effect that enables the development of microchips for various uses is achieved.

(Embodiment 4)
In the present embodiment, a method for forming a structure in one flow path will be described. FIG. 7A and FIG. 7B show an example of a flow path in which a structure is formed, and is a cut surface parallel to the fluid moving direction.

FIG. 7A shows a flow channel 51 in which a stepped structure 81 is formed inside one flow channel. The flow path 51 has a shape in which the flow path becomes narrower in stages with respect to the moving direction of the fluid. By using the flow path 51, for example, the flow rate and pressure of the fluid can be controlled.

FIG. 7B shows a flow channel 53 in which a plate-like structure 83 is formed inside one flow channel. The flow path 53 has a shape in which plate-like structures are arranged at predetermined intervals with respect to the direction of fluid movement, and the flow path becomes narrower in stages. By using the flow path 53, for example, fluid can be stirred. When two or more kinds of fluids are introduced into the flow path 53, the introduced fluids are agitated by colliding with the plate-like structure 83. The stirring effect can be enhanced by forming the plurality of plate-like structures 83 at a predetermined interval inside one flow path.

In order to form the structure of the present embodiment inside one flow path, the flow path forming method described in the second embodiment can be applied. In order to form a flow channel structure by a conventional semiconductor manufacturing technique, a complicated manufacturing process is required. By using the structure forming method according to the present embodiment, a complicated flow path can be easily formed as compared with the conventional method. Therefore, the use of the flow path having the structure of the present invention according to the present embodiment increases the degree of freedom in designing a microchip having functionality in the flow path, and development of microchips for various applications. It has an excellent effect that makes it possible.

(Embodiment 5)
In the present embodiment, an example in which the method for forming a flow path having a complicated shape described in the second embodiment is applied to the manufacture of a microchip will be described. FIG. 8 shows a microchip 200 according to this embodiment of the present invention. FIG. 8A is a view of the microchip 200 as viewed from above, FIG. 8B is a cut surface at AA ′, and FIG. 8C is a cut surface at BB ′. For example, the microchip 200 has a flow channel 261 having a micro pillar 285 inside a single substrate 1, and the flow channel 211 includes a flow channel 261 and an opening 290 formed on the upper surface of the substrate 1. It is connected to the. The opening 290 may be formed on the side surface of the microchip 200. The micro pillar 285 is a cylindrical structure and is formed integrally with the bottom surface of the substrate 1.

The microchip 200 of the present embodiment uses the relationship between the irradiation method of the laser beam 5 described above and the shape of the modified portion 7 to be formed, so that the laser beam propagates through the previously formed modified portion 7. 5 is scanned to form a relatively large reforming portion 7 of the flow path 261 and a micro pillar 285 or a narrow flow path 211 is formed, so that it does not propagate through the reforming section 7 previously formed. Further, it can be formed by fine processing by scanning the focal point 6 of the laser beam 5.

When the microchip 200 having the micro pillar 285 is manufactured by a conventional semiconductor manufacturing technology, the micro pillar 285 is formed by anisotropic etching, and then the upper surface portion of the base material 1 is bonded to form the manufacturing process. Becomes complicated. In addition, since a plurality of base materials are used, the manufacturing cost increases. However, by using the flow path forming method according to the present embodiment, a microchip can be formed with a single substrate, and can be more easily formed as compared with the conventional case.

Specific examples of the microchip using the flow channel according to the present invention described in the above embodiment are shown below. In addition, the following Examples are examples and the microchip using the flow path which concerns on this invention is not limited to this.

Example 1
FIG. 9 is a schematic diagram of a microchip 300 according to the present invention of Example 1, FIG. 9 (a) is a top view, and FIG. 9 (b) is a cross-sectional view at BB ′ in FIG. 9 (a). . The microchip 300 has a channel 311 inside a single substrate 1 and a plurality of protruding traps 315 in the middle of the channel 311, and an opening 390 a connected to the channel 311 on the upper surface of the substrate 1. And an opening 390b. In addition, a part of the bottom surface of the trap 315 penetrates, and the trap 315 is closed by forming an electrode 395 on the bottom surface. In FIG. 9A, the flow path 311 is formed as a meandering flow path. However, the flow path 311 can be arranged in a shape according to the purpose, such as a straight line, a curved line, or a spiral.

As the substrate 1, borosilicate glass, soda lime, alumina borosilicate glass, quartz glass, or the like can be used, and Pyrex (registered trademark) glass excellent in corrosion resistance can be suitably used. For the electrode 395, a metal such as aluminum or platinum can be used.

The microchip 300 can be used for observation of a particulate test object such as a cell, for example. When the electrode 395 disposed in the trap 315 is irradiated with laser light, the electrode 395 vibrates and bubbles can be generated. As a result, the particulate test object that has entered the trap 315 is pushed out to the flow path 311 and becomes a driving force for the fluid that fills the flow path 311. When a fluid containing a test object is introduced into the flow path 311 from the opening 390a, the fluid moves in the direction of the opening 390b. At this time, as described above, the electrode 395 is irradiated with laser light to generate bubbles, thereby facilitating the movement of the fluid and observing the test object moving in the flow path 311. The electrode 395 can also be used to measure a test object that has entered the trap 315 by an electric resistance method.

The microchip of this example meandered into the inside of a single substrate by combining pulsed laser irradiation and wet etching without using a conventional method of bonding or stacking the substrates. Channels, traps, etc. can be formed. Since the microchip of this example is formed of a single glass substrate, it is suitable for observing a test object that moves in the flow path.

(Example 2)
10A and 10B are schematic views of a microchip 400 according to the present invention of Example 2. FIG. 10A is a top view, and FIG. 10B is a cross-sectional view taken along BB ′ in FIG. . The microchip 400 has a relatively large rectangular channel 411 inside a single substrate 1, and an opening 490 a and an opening 490 b connected to the channel 411 on the upper surface of the substrate 1. In addition, the flow path 411 includes a plurality of micro pillars 485 that are columnar structures formed integrally with the base material 1 on the bottom surface of the flow path 411 and a plurality of micro pockets 487 including two triangular pillars. Electrodes 495a, 495b, 495c, and 495d are disposed on the bottom surface of the microchip 400.

As the substrate 1, borosilicate glass, soda lime, alumina borosilicate glass, quartz glass, or the like can be used, and Pyrex (registered trademark) glass excellent in corrosion resistance can be suitably used. For the electrode 395, a metal such as aluminum or platinum can be used.

The microchip 400 can be used for observation of chromosomal DNA, for example. The observation method will be described below. FIG. 11 is a diagram showing a method for observing chromosomal DNA using the microchip 400. First, a solution in which the cells 1101 are suspended is introduced into the channel 411 from the opening 490a. The introduced cell 1101 diffuses in the channel 411, and excess solution is discharged out of the microchip through the opening 490b. As the solution for suspending the cells 1101, physiological saline or PBS (phosphate buffered saline) is suitable for optical observation.

Here, when a positive potential is applied to the electrode 495a and a negative potential is applied to the electrode 495b, the cell 1101 moves to the electrode 495b as shown in FIG. The micro pocket 487 is disposed so as to accommodate exactly one cell between two triangular prisms, and the cell 1101 is accommodated in the micro pocket 487. Since the microchip 400 is formed of transparent glass, the movement of the cells 1101 can be easily observed by using a phase contrast microscope.

Next, a solubilizer is added from the opening 490a to the channel 411 to lyse the cells 1101. As the solubilizer, a surfactant such as SDS (sodium lauryl sulfate) or a denaturing agent such as guanidine hydrochloride can be used. Moreover, you may add proteolytic enzymes, such as trypsin, in order to decompose | disassemble a protein. The cell membrane of the cell 1101 accommodated in the micro pocket 487 is dissolved by the solubilizer, and the chromosomal DNA 1103 is released from the cell 1101.

When a negative potential is applied to the electrode 495a and a positive potential is applied to the electrode 495b, the chromosomal DNA 1103 extends toward the electrode 495a as shown in FIG. Further, when a positive potential is applied to the electrode 495c and a negative potential is applied to the electrode 495d, the chromosomal DNA 1103 is adsorbed on the surface of the micropillar 485, as shown in FIG.

As described above, when the chromosomal DNA is extended outside the cell, the chromosomal DNA can be easily observed using the FISH (Fluorescence In-Situ Hybridization) method. The FISH method is an observation method of a base sequence using a fluorescent DNA probe having a base sequence complementary to a target base sequence. Usually, since the FISH method is used for condensed chromosomal DNA, the resolution during observation is not high. However, by using the microchip 400 of this embodiment, the chromosomal DNA can be extended outside the cell, and the target base sequence can be observed with high resolution and high accuracy by the FISH method. .

In the microchip of this example, a combination of pulsed laser irradiation and wet etching is used in combination with a base material without using a conventional base material laminating or laminating manufacturing method. And a channel having a micro pocket can be formed. In addition, since the microchip of this example is formed of a single glass substrate, it is suitable for observing the target base sequence by the FISH method.

(Example 3)
12A and 12B are schematic views of a microchip 500 according to the present invention of Example 3. FIG. 12A is a top view, and FIG. 12B is a cross-sectional view taken along BB ′ in FIG. . The microchip 500 has a flow path 511 that meanders inside a single substrate 1, and an opening 590 a, an opening 590 b, an opening 590 c, and an opening 590 d that are connected to the flow path 511 on the upper surface of the substrate 1. . The flow path 511 includes a pretreatment unit 1251, a reaction unit 1253, and a detection unit 1255 from the opening 590a side. Further, a plate-like structure 583 formed integrally with the base material 1 on the bottom surface of the flow channel 511 is provided inside the flow channel 511.

As the substrate 1, borosilicate glass, soda lime, alumina borosilicate glass, quartz glass, or the like can be used, and Pyrex (registered trademark) glass excellent in corrosion resistance can be suitably used.

The microchip 500 can be used, for example, in a detection kit using an ELISA (Enzyme-Linked Immunosorbent Assay) method. A method for detecting an object to be detected using the ELISA method will be described below. FIG. 12 is a diagram showing a method for detecting an object to be detected by the ELISA method using the microchip 500. FIG. First, the solution to be detected is introduced into the flow path 511 from the opening 590a. The introduced solution moves in the channel 511, and excess solution is discharged out of the microchip through the opening 490d.

When the object to be detected is intracellular protein or the like, a solubilizer is added from the opening 590a to lyse the cell. As the solubilizer, a surfactant such as SDS (sodium lauryl sulfate) or a denaturing agent such as guanidine hydrochloride can be used.

In order to remove unnecessary components in the solution, a filter or an ion exchange resin may be provided in the pretreatment unit 1251. In this case, the detection object 1205 solubilized in the solution is roughly purified, so that the detection sensitivity can be improved. The detected object 1205 that has moved through the flow path 511 is fixed by the immobilized beads 1211 that have been introduced into the reaction unit 1253 in advance. The immobilized beads 1211 are fastened to the reaction unit 1253 by the plate-like structure 583. Subsequently, a cleaning liquid is introduced into the flow path 511 from the opening 590b, and components that are not fixed to the immobilized beads 1211, here, components other than the detection target 1205 are washed away. As the washing solution, physiological saline or PBS (phosphate buffered saline) is suitable for optical observation.

Next, the antibody 1207 is added to the flow path 511 from the opening 590b. The antibody 1207 is an antibody that specifically binds to the detection target 1205, and may be a polyclonal antibody or a monoclonal antibody. Here, the antibody 1207 labels the labeling enzyme 1209. Subsequently, a cleaning solution is introduced into the flow path 511 from the opening 590b, and excess antibody 1207 not bound to the detection target 1205 is washed away.

Next, the substrate 1213 is added from the opening 590c. The substrate 1213 becomes a reaction product 1215 by the labeling enzyme 1209 of the antibody 1207 bound to the detection target 1205. The reaction product 1215 is detected at the detection position 1260 of the detection unit 1255.

By generating the reaction product 1215 from the substrate 1213, it is indirectly detected that the detection target 1205 is present in the solution. The reaction product 1215 can be detected optically, and a tendency microscope, a thermal lens microscope, or the like can be used.

The microchip of this example has a plate-like shape inside a single substrate by combining pulsed laser irradiation and wet etching without using a conventional method of bonding or laminating substrates. A flow path having a structure can be formed. In addition, since the microchip of this example is formed of a single glass substrate, it is suitable for detection by the ELISA method.

1: substrate 5: laser 6: focus 7: reforming part 11: flow path 21: flow path 23: flow path 31: flow path 33: flow path 41: flow path 43: flow path 51: flow path 53: flow Road 61: Channel 81: Step-like structure 83: Plate-like structure 85: Micro pillar 90: Opening portion 100: Microchip 200: Microchip 211: Channel 261: Channel 285: Micro pillar 290: Opening 300 : Microchip 311 Channel 315: Trap 390a: Opening 390b: Opening 395: Electrode 400: Microchip 411: Channel 485: Micro pillar 487: Micro pocket 490a: Opening 490b: Opening 495a: Electrode 495b: Electrode 495c: Electrode 495d: Electrode 500: Microchip 511: Channel 583: Plate-like structure 590a: Opening 590b: Opening 59 c: opening 590D: opening 1101: Cell 1103: DNA
1205: Detected object 1207: Antibody 1209: Label enzyme 1211: Bead 1213: Substrate 1215: Reaction product 1251: Pretreatment unit 1253: Reaction unit 1255: Detection unit 1260: Detection position

Claims (10)

Borosilicate glass, soda lime, alumina borosilicate glass, in the interior of a single transparent substrate selected from quartz glass and sapphire, have at least two openings in the surface or the side surface of the substrate, wherein the group have a flow path that is formed to bend in the horizontal and / or vertical direction with respect to the upper surface of the base material in the wood,
Microchip, characterized in that the perforated projecting from a surface of said flow path, said substrate and integrally formed a plate-like or columnar structure in the flow path. After being modified by laser, the substrate is formed by isotropic etching inside a single transparent substrate selected from borosilicate glass, soda lime, alumina borosilicate glass, quartz glass and sapphire , and have at least two openings in the surface or side of the timber, have a flow path that is formed to bend in the horizontal and / or vertical direction with respect to the upper surface of said substrate in said substrate,
Microchip, characterized in that the perforated projecting from a surface of said flow path, said substrate and integrally formed a plate-like or columnar structure in the flow path. 3. The microchip according to claim 1, wherein a cross-sectional shape of the channel is a circle, an ellipse, a polygon, a ring, or a donut. The microchip according to any one of claims 1 to 3 , wherein a cross-sectional shape of the flow path varies depending on a position of the base material. A plurality of the structures in the flow path, the microchip according to any one of claims 1 to 4 wherein the plurality of structures are characterized by having a different structure sizes. Irradiating laser inside a single substrate selected from borosilicate glass, soda lime, alumina borosilicate glass, quartz glass and sapphire to modify a part of the substrate,
And a portion of the substrate is removed by etching, at least two have a opening, horizontal and / or vertical direction with respect to the upper surface of the substrate within the substrate surface or side surface of the substrate A flow path formed to bend in the base material ,
A microchip manufacturing method , wherein a plate-like or columnar structure that protrudes from a surface of the flow path and is formed integrally with the base material is formed in the flow path . The microchip manufacturing method according to claim 6 , wherein the laser is a pulse laser of 1000 femtoseconds or less. 8. The method of manufacturing a microchip according to claim 7 , wherein a cross-sectional shape of the flow path is formed in a circular shape, an elliptical shape, a polygonal shape, a ring shape, or a donut shape. 9. The method of manufacturing a microchip according to claim 8 , wherein a cross-sectional shape of the flow path is formed to be different depending on a position of the base material. Microchip manufacturing method according to any one of claims 6 to 9, characterized by forming a plurality of different structures sizes in the flow path.