JPH1076168A - Pipette and its production as well as production of capillary - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a pipette with which the deformation of an operation object by operation may be lessened even if the operability of the operation object is good and the operation object is easily deformable. SOLUTION: This pipette has a capillary part 10 having the opening at the front end to be used as a suction port 10a and a cup-shaped housing part 11. The housing part 11 is formed integrally with the capillary part 10 at the front end of the capillary part 10 and forms the housing space 11a of the operation object 1 by enclosing the circumference of the operation object 1 sucked and held at the suction port 10a from the lateral side with respect to the suction direction of the operation object 1 by the suction port 10a.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a pipette for manipulating minute objects such as minute particles such as cells and minute mechanical parts.

[0002]

2. Description of the Related Art Conventionally, a glass pipette as a holding tool has been used for manipulating minute objects such as protozoa and egg cells. FIG. 10 is a sectional view showing a main part of this conventional pipette. As shown in FIG. 10, this conventional pipette has a suction port 2 opened to the outside to suck and hold an operation target 1 such as a cell, and a connection port opened to the outside to draw a negative pressure (not shown). ) And an internal passage 3 for communicating between the suction tube 2 and the connection port. Note that the connection port is a suction port 2 in the glass tube 4.
The opening of the internal passage 3 on the opposite side from the opening.

According to this pipette, the connection port side portion of the glass tube 4 is connected to, for example, a so-called injector (configured like a syringe), and the injector is operated to operate the internal passage through the connection port. A negative pressure is applied to 3.
As a result, as shown in FIG. 10, the operation target 1 such as a cell is suction-held at the suction port 2. In this state, for example, when the operation target 1 is an egg cell, a glass capillary (glass tube for injecting sperm) 5 is inserted into the egg cell to inject sperm to perform fertilization. In the case of a cell or the like, various operations such as making a hole in the operation target 1 with a needle or cutting with a cutting blade (not shown) are performed. After such an operation is completed, the injector is operated to operate the internal passage 3 through the connection port.
The inside is made to have a positive pressure, and the suction holding state of the operation target 1 by the suction port 2 is released.

The conventional method for manufacturing a pipette will be described. For example, a glass tube having a diameter of 1 to 2 mm is stretched while being heated by a needle puller (not shown). Thereby, the tube is thinned and cut, and two glass tubes 4 can be obtained. The opening at the cut end becomes the suction port 2 of the pipette. However, since the tip of the pipette is usually sharp as it is when cut, the operation target 1 is damaged if used as it is. Therefore, the tip of the glass tube 4 is heated by a device such as a microforge (not shown), Apply rounding process.

[0005]

However, the conventional pipette has a drawback that the operability of the operation target 1 is extremely poor. That is, for example, as shown in FIG. 10, the cell to be operated 1 is held by the conventional pipette 4, fertilization is performed by inserting a glass capillary 5 into the cell, or a hole is formed by piercing the cell with a needle, When an operation such as cutting or incising a cell with a cutting blade is performed, the cell is held at only one point of the suction port 2, and the cell swings around this point. The first operation has a drawback that requires a great deal of labor. When the operation target 1 is a cell, since the cell is a viscoelastic substance, the cell is greatly deformed by an external force, and the deformation may affect the active state of the cell. Furthermore, when cutting or incising cells with a cutting blade, not only the cells swing as described above, but also as a cutting blade, for example, a metal blade,
Since a very thin plate-shaped material such as a thin film plate formed by using a semiconductor manufacturing technique disclosed in Japanese Patent Application Laid-Open No. 8-85018 is used, the cutting blade is liable to move in the thickness direction. It is extremely difficult to cut or incise cells and the like along a desired plane.

Further, since the conventional pipette is manufactured by the above-described manufacturing method, the conventional pipette is manufactured by pulling and heating a glass tube according to personal experience. The skilled artisan is required to produce pipettes of the same shape. In addition, the conventional pipette requires a lot of trouble in manufacturing, and the cost is unavoidable. Further, since the size of the suction port 2 of the conventional pipette is determined when the glass tube is cut, there is a problem that it is very difficult to obtain a pipette having the suction port 2 having a desired size. ing. Note that the desired size of the suction port 2 is different depending on various operation targets 1.

The present invention has been made in view of the above circumstances, and provides a pipette capable of reducing the deformation of the operation target due to the operation even if the operation target is easily deformable even if the operation target is easy to deform. The purpose is to provide.

It is another object of the present invention to provide an inexpensive pipette having an arbitrary size and a high-precision suction port, which is easy and inexpensive to manufacture, and a method for manufacturing the pipette.

Another object of the present invention is to provide a pipette that can easily cut or incise an operation target along a desired plane.

Still another object of the present invention is to provide a method for producing a thin tube which can be used in such a method for producing a pipette.

[0011]

According to a first aspect of the present invention, there is provided a pipette according to a first aspect of the present invention, comprising: a thin tube having an opening at a distal end serving as a suction port; A cup-shaped housing portion integrally formed and surrounding the operation target sucked and held by the suction port from a side in a suction direction of the operation target by the suction port to form a storage space for the operation target, It is provided with. The accommodation space formed by the accommodation section does not necessarily need to accommodate all of the operation objects, and may accommodate, for example, about half of the operation objects.

According to the first aspect, even if the operation target sucked and held by the suction port attempts to swing around the suction port due to an external force, the periphery of the operation target is restrained by the inner wall of the housing, The swing is almost suppressed, the operation of the operation target is facilitated, and the operability of the operation target is improved. Further, even if the operation target sucked and held by the suction port is about to be deformed by an external force, the periphery of the operation target is restrained by the inner wall of the housing portion, and the deformation is almost suppressed.

[0013] A pipette according to a second aspect of the present invention is the pipette according to the first aspect, wherein the thin tube portion and the housing portion are integrally formed of silicon oxide.

In the pipette according to the second aspect, since the thin tube portion and the housing portion are integrally formed of silicon oxide, the pipette can be manufactured by a method as described later in sixth to eighth aspects. In addition to having a suction port with a size of high accuracy and high precision, it is easy and inexpensive to manufacture. Further, since silicon oxide is transparent, the thin tube portion and the housing portion become transparent. Therefore, since the housing portion is transparent, the suction port and the operation target sucked and held by the suction port can be observed under reflected light or transmitted light under a microscope or the like, and the operation of the operation target can be easily performed. Become. Further, since the thin tube portion is also transparent, the thin tube portion does not obstruct the observation light at the time of the observation, and the observation becomes easy.

A pipette according to a third aspect of the present invention is the pipette according to the first or second aspect, wherein the pipette is a support for supporting a base end of the thin tube portion, and is opened to the outside to draw a negative pressure. A support having a connection port for, and an internal passage communicating between the inside of the thin tube portion and the connection port,
It is further provided.

According to the third aspect, since the base end of the thin tube is supported by the support, the support can be held instead of the thin tube in use. For this reason, holding of the pipette is facilitated, and there is no possibility of damaging the thin tube portion.

A pipette according to a fourth aspect of the present invention is the pipette according to any one of the first to third aspects, wherein a guide blade for cutting the object to be operated is introduced into the housing space and the cutting edge is provided. A guide slit for restricting the movement of the guide member substantially within a predetermined plane is formed in the housing portion.

According to the fourth aspect, when cutting or incising a cell or the like as an operation target sucked and held by the suction port, the guide slit is formed in the housing portion. By inserting a cutting blade into the guide slit, the cutting blade can be guided or guided by the guide slit to cut or cut the held cells or the like. As described above, not only the swing of the cells and the like is suppressed, but also the movement of the cutting edge is substantially restricted to a predetermined plane by the guide slit, so that the deflection of the cutting edge is also suppressed. Therefore, cells or the like can be easily cut or incised along a desired plane.

The cutting blade may be, for example, a metal blade or a thin plate of a micro-cutting device disclosed in Japanese Patent Application Laid-Open No. 8-85018.

The pipette according to a fifth aspect of the present invention is the pipette according to the fourth aspect, wherein the guide slit is provided in the housing section so that the cutting blade can be inserted simultaneously across the housing space. It is formed on both sides of the accommodation space.

According to the fifth aspect, if the cutting blade is inserted into the guide slits on both sides of the accommodation space while straddling the accommodation space, the cutting edges are formed by the guide slits on both sides of the accommodation space. Of the cutting blade is more reliably suppressed, and cells or the like can be more easily cut or incised along a desired plane. However, in the first to fourth aspects, the guide slit may be formed only on one side of the accommodation space in the accommodation part.

A pipette manufacturing method according to a sixth aspect of the present invention is a pipette manufacturing method for manufacturing the pipette according to any one of the first to fifth aspects, wherein the first and second pipettes are manufactured.
A silicon substrate, wherein at least one surface of the first and second silicon substrates has a recess including a portion corresponding to at least the capillary portion and a portion corresponding to the housing portion. Preparing the formed first and second silicon substrates, and forming a cavity by the recess in a joined body of the first silicon substrate and the second silicon substrate; Bonding the first silicon substrate and the second silicon substrate, thermally oxidizing a wall surface of the cavity, and at least a portion of the first and second silicon substrates that is not thermally oxidized. And removing a portion by etching.

According to the sixth aspect, since a pipette can be manufactured by using a semiconductor manufacturing technique, a pipette having an arbitrary shape and a highly accurate suction port can be easily manufactured. It is also possible to manufacture a plurality of pipettes simultaneously and collectively, and it is possible to manufacture pipettes at a lower cost.

In a pipette manufacturing method according to a seventh aspect of the present invention, in the pipette manufacturing method according to the sixth aspect, after the joining step and before the thermal oxidation step, the hollow portion is externally mounted. And forming a groove in the joined body communicating with the joint.

According to the seventh aspect, since the cavity communicates with the outside by the groove, oxygen is easily supplied into the cavity at the time of thermal oxidation. Therefore, thermal oxidation of the inner wall of the cavity is prevented. Accelerated and preferred. However, if the recess is formed in advance in the silicon substrate so that the cavity communicates with the outside in the first place, and oxygen is sufficiently supplied into the cavity, the recess is not necessarily required. It is not necessary to form a groove for communicating the cavity with the outside as in the case of the first embodiment.

The pipette manufacturing method according to an eighth aspect of the present invention is the pipette manufacturing method according to the sixth or seventh aspect, wherein the thermal oxidation in the first and second silicon substrates is performed after the thermal oxidation step. At least a part of the portion that does not correspond to the thin tube portion and the housing portion of the portion that has been removed is removed or the thin tube portion of the thermally oxidized portion of the first and second silicon substrates is removed. And a step of making the portion corresponding to the storage portion discontinuous. Depending on the condition of the formation of the recess in the silicon substrate, there is a wasteful or obstructive portion in the thermally oxidized portions of the first and second silicon substrates, which does not correspond to the thin tube portion and the accommodating portion. There is. In such a case, it is preferable to remove or discontinue the wasteful or obstructive portion from the portion corresponding to the thin tube portion and the housing portion as in the eighth aspect. Note that this removal or discontinuity is achieved by, for example, making a cut in a predetermined portion of the bonded body of the first and second silicon substrates after the thermal oxidation using a dicing saw or the like (that is, forming a groove). be able to. In the case of discontinuity, even if both are connected via a silicon portion, they may be separated at the stage of removing by etching or the like.

According to a ninth aspect of the present invention, there is provided a method of manufacturing a thin tube, comprising: a first and a second silicon substrate; and a concave portion formed on at least one surface of the first and the second silicon substrate. And a recess including at least a portion corresponding to the capillary,
Preparing the formed first and second silicon substrates, and forming a cavity by the recess in a joined body of the first silicon substrate and the second silicon substrate; Bonding the first silicon substrate and the second silicon substrate, thermally oxidizing a wall surface of the cavity, and at least a portion of the first and second silicon substrates that is not thermally oxidized. And removing a portion by etching.

The method for producing a thin tube according to the ninth aspect is as follows.
It is used in the pipette manufacturing method according to the sixth to eighth aspects (particularly when forming a thin tube portion). However, the thin tube manufactured according to the ninth aspect can be used for other applications such as a micromachine. According to the ninth aspect, since a thin tube can be manufactured by using a semiconductor manufacturing technology, a plurality of high-precision fine thin tubes can be simultaneously manufactured at once.

[0029]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a pipette according to the present invention, a method for manufacturing the same, and a method for manufacturing a thin tube will be described in detail with reference to the drawings.

First, a pipette according to an embodiment of the present invention will be described with reference to FIG. FIG. 1 is a diagram schematically showing a pipette according to the present embodiment.
1A is a schematic perspective view thereof, and FIG. 1B is a schematic cross-sectional view of a main part thereof. For convenience of explanation, X, Y, and Z coordinate systems orthogonal to each other are defined as shown in FIG.

As shown in FIG. 1, the pipette according to the present embodiment has a thin tube portion 10 in which an opening 10a at the distal end serves as a suction port.
And the direction of suction of the operation target by the suction port 10a around the operation target 1 formed integrally with the thin tube portion 10 at the tip of the thin tube portion 10 and suction-held by the suction port 10a (FIG. 1B).
(A right direction in the middle) from the side to form a housing space 11a for the operation target 1 and a cup-shaped housing portion 11. The accommodating portion 11 has a size enough to accommodate the desired operation target 1 (a size enough to accommodate the entire operation target 1 with some margin, for example, a half of the operation object 1 with some margin). It may be of a size that can be accommodated).

In this embodiment, the thin tube portion 10 and the housing portion 11 are integrally formed of silicon oxide.

Further, as shown in FIG. 1, the pipette according to the present embodiment has a support 1 for supporting the proximal end of the thin tube portion 10.
2 is provided. The support 12 has a connection port 12a that opens to the outside to draw a negative pressure, and an internal passage 12b that allows communication between the inside of the thin tube portion 10 and the connection port 12a.

According to the pipette according to the present embodiment, the portion of the support 12 on the side of the connection port 12a is connected to, for example, an injector, and the injector is operated to operate the inside of the internal passage 12a and the inside of the thin tube section 10 through the connection port 12a. To a negative pressure.
As a result, as shown in FIG. 1B, the operation target 1 such as a cell is sucked and held by the suction port 10a, and is housed in the housing space 11a formed by the housing unit 11. In this state, for example, when the operation target 1 is an egg, a glass capillary (glass tube for injecting sperm) 5 is inserted into the egg to inject sperm to perform fertilization. In the case of a cell or the like, various operations are performed, such as making a hole in the operation target 1 with a needle or the like. At this time, even if the operation target 1 sucked and held by the suction port 10a attempts to swing around the suction port 10a due to an external force, the periphery of the operation target 1 is restrained by the inner wall of the housing portion 11, and the swing is hardly caused. It is suppressed, the operation of the operation target 1 is facilitated, and the operability of the operation target 1 is improved. In addition, suction port 1
Even if the operation target 1 sucked and held at 0a attempts to deform due to an external force, the periphery of the operation target 1 is restrained by the inner wall of the housing 11, and the deformation is almost suppressed. After such an operation is completed, the injector is operated to operate the internal passage 12b and the thin tube section 10 through the connection port 12a.
Is set to a positive pressure, and the suction holding state of the operation target 1 by the suction port 10a is released.

Further, according to the pipette according to the present embodiment, since the thin tube portion 10 and the housing portion 11 are made of silicon oxide, and the silicon oxide is transparent, the thin tube portion 10 and the housing portion 11 are made transparent. Become. Therefore, since the housing portion 11 is transparent, the suction port 10a and the operation target 1 held by the suction port 10a can be observed under reflected light or transmitted light under a microscope or the like. Operation becomes easy. Further, since the thin tube portion 10a is also transparent, the thin tube portion 10 does not obstruct the observation light at the time of the observation, and the observation becomes easy. However, in the present invention,
The material forming the thin tube portion 10 and the housing portion 11 is not necessarily limited to silicon oxide.

Further, in the present embodiment, since the base end of the thin tube portion 10 is supported by the support 12, the support 12 can be held instead of the thin tube 10 in use. For this reason, holding of the pipette, connection with the injector, and the like are facilitated, and there is no possibility of damaging the thin tube portion 10. However, in the present invention, the support 12 is not always necessary.

Next, an example of a method of manufacturing the pipette shown in FIG. 1 will be described with reference to FIGS. FIG. 2 is a cross-sectional view showing each step of the method for manufacturing the pipette shown in FIG. FIG. 3 is a cross-sectional view showing each step following the step shown in FIG. 2 of the method for manufacturing the pipette shown in FIG. The cross sections shown in FIGS. 2 and 3 correspond to YZ in FIG.
Corresponds to the cross section. FIG. 4 is a plan view showing a predetermined step shown in FIG. 2, and FIG. 4 (a) is AA ′ in FIG. 2 (a).
FIG. 4B is an arrow view, FIG. 4B is an arrow view taken along the line BB ′ in FIG.
FIG. 4C is a view taken along the line CC ′ in FIG. 2D.

First, a silicon substrate 20 having a diameter of 3 inches and a thickness of 400 μm and having a (100) plane orientation was used as a substrate material, and dichlorosilane and ammonia gas were used as mask materials on both surfaces of the substrate 20 by low-pressure vapor deposition. Then, silicon nitride films 21 and 22 are formed to a thickness of 100 nm. Further, by patterning the silicon nitride film 21 on the upper surface of the substrate 20 by a photolithography method and a dry etching method, a plurality of openings 21a for exposing the upper surface of the substrate 20 are formed at predetermined positions of the silicon nitride film 21 on the upper surface of the substrate 20. Form. The size of the opening 21 is, for example, 150 μm × 2
It is 50 μm. Thereafter, the substrate is immersed in an etching solution for silicon such as an aqueous solution of potassium hydroxide (KOH) or an aqueous solution of tetramethylammonium hydroxide (TMAH), and the substrate exposed from the opening 21a using the silicon nitride films 21 and 22 as masks. By etching the portion 20, a plurality of recesses 23 having a depth of 50 μm are simultaneously formed on the upper surface of the substrate 20. (FIG. 2 (a), FIG. 4 (a)). Since the substrate 20 has a (100) plane orientation, as is well known, etching is performed on silicon (1).
11) Since the surface of the recess 23 is automatically stopped at the surface, the surface of the recess 23 becomes a taper surface having a predetermined angle. In the present embodiment, one recess 23 includes a portion corresponding to two storage portions 11 corresponding to two pipettes.

Next, a recess having a depth of 10 μm including a portion corresponding to the narrow tube portion 10 (this embodiment) is formed in the same manner as the recess 23 is formed after the silicon nitride films 21 and 22 are once removed. In the embodiment, a plurality of grooves 24 are formed on the upper surface of the substrate 20 (FIGS. 2B and 4B). The recess 23 and the corresponding recess 24 are continuous. In this case, the silicon nitride film remains on both surfaces of the substrate 20, but the silicon nitride film (not shown) on the upper surface is removed, and only the silicon nitride film 25 on the lower surface remains. FIGS. 2B and 4B show this state.

The recess 23 including the portion corresponding to the housing portion 11 and the recess 24 including the portion corresponding to the thin tube portion 10 are
It may be formed by dry etching or the like instead of wet etching, or may be formed by dicing (mechanical processing using a dicing saw or a processing machine similar thereto).

Two silicon substrates 20 in the state shown in FIGS. 2 (b) and 4 (b) are prepared, and a cavity formed by the recesses 23 and 24 is provided inside a joined body of one substrate and the other substrate. Then, the two substrates are joined to each other so as to form (FIG. 2C). In the present embodiment, two silicon substrates 2
The surfaces on which the 0 recesses 23 and 24 are formed are joined. This bonding is performed by, for example, about 1
It is performed at a temperature of 200 ° C.

Thereafter, a groove 26 is formed in the joined body shown in FIG. 2C so that the cavity formed by the recesses 23 and 24 communicates with the outside (FIGS. 2D and 4C). In the present embodiment, a groove 26 having a width of 30 μm is formed by dicing to reach the recess 23 from the upper surface of the joined body. The groove 26 is processed linearly as shown in FIG. 4C, but is aligned so as to penetrate the plurality of recesses 23 by one processing. This groove 26
It is formed in order to sufficiently supply oxygen into the cavity in a later-described thermal oxidation treatment of the inner wall of the cavity. However, if the recesses 23 and 24 are previously formed in the silicon substrate 20 so that the cavity communicates with the outside in the first place, and oxygen is sufficiently supplied into the cavity, the groove is not necessarily required. It is not necessary to form 26.

Next, the joined body in the state shown in FIG. 2D and FIG. 4C is placed in an electric furnace, and the inner wall (silicon portion) of the cavity formed by the recesses 23 and 24 is thermally oxidized (FIG. 3).
(A)). At this time, the inner wall of the groove 26 is also thermally oxidized. Reference numeral 27 in FIG. 3A indicates a silicon oxide film formed by thermal oxidation. The thickness of the silicon oxide film 27 is
For example, it is about 2 μm.

Next, a groove having a width of 100 μm is used to separate the central portion of the recess 23 along the groove 26 by using a blade wider than the processing blade used for dicing for forming the above-described groove 26. 28 are formed (FIG. 3B). As a result, the silicon oxide film 27 formed on the inner wall of one recess 23 is separated into ones corresponding to individual pipettes, and portions of the silicon oxide film 27 that do not correspond to the thin tube portion 10 and the housing portion 11. Groove 2 which is unnecessary part
The portion formed on the inner wall of No. 6 is removed.

Thereafter, the silicon nitride films 25 on the upper and lower surfaces of the joined body are formed.
Is patterned according to a desired shape of the support 12 by a photolithography method and a dry etching method (FIG. 3C).

Finally, the conjugate in the state shown in FIG. 3C was replaced with tetramethylammonium hydroxide (TMA).
H) Immerse in an aqueous solution (this aqueous solution hardly etches the silicon oxide film) and remove unnecessary silicon portions (support 2
Is removed by etching (FIG. 3D). Thereby, a plurality of pipettes shown in FIG. 1 are completed collectively.

Further, since the pipette according to the present embodiment can be manufactured by using the semiconductor manufacturing technique as described above, it is possible to obtain the suction port 10a having an arbitrary shape and high accuracy, and It is also possible to manufacture a plurality of pipettes simultaneously and collectively, and it is possible to manufacture pipettes at a lower cost.

In the above-described manufacturing method, the bonding 2
Although the recess including the portion corresponding to the thin tube portion and the portion corresponding to the housing portion is formed on both of the two silicon substrates 20, in the present invention, the recess may be formed only on one substrate. Good. Even in this case, a cavity having an inner wall of silicon can be formed in the joined body of the two substrates.

If attention is paid only to the thin tube portion 10 of the pipette, the above-described manufacturing method includes a method of manufacturing a thin tube, and it is difficult to manufacture an arbitrary thin tube according to the above-described manufacturing method. it is obvious.

Next, a pipette according to another embodiment of the present invention will be described with reference to FIG. FIG. 5 schematically shows a pipette according to the present embodiment.
FIG. 5A is a schematic perspective view, and FIG. 5B is a schematic sectional view of a main part thereof. For convenience of explanation, X, Y, and Z coordinate systems orthogonal to each other are defined as shown in FIG.

In FIG. 5, the same or corresponding elements as those in FIG. 1 are denoted by the same reference numerals, and description thereof will be omitted.

The point that the pipette according to the present embodiment is different from the pipette shown in FIG. 1 described above is to guide the cutting blade 30 for cutting the operation target 1 into the accommodation space 11a and to substantially move the cutting blade 30. A predetermined surface (in the present embodiment, FIG. 1
(B) a guide slit 31 that is regulated in a plane including the normal to the paper surface and the central axis of the housing 11 and the thin tube 10 in FIG. 1). 12 is only configured to be narrow.

The slit width of the guide slit 31 is determined according to the thickness of the cutting blade 30 to be used.
The length of 1 is determined according to a desired cutting amount for the operation target 1 such as a cell. When cutting cells, there are a case where cells are completely cut and a case where cells are cut halfway and the outer skin of the cells is not separated.

In the present embodiment, the guide slit 31
Are formed on both sides of the housing space 11a so that the cutting blade 30 can be inserted simultaneously across the housing space 11a.
Therefore, if the cutting blade 30 is inserted into the guide slits 31 on both sides of the housing space 11a while straddling the housing space 11a, the movement of the cutting blade 30 is regulated by the guide slits 31 on both sides of the housing space 11a. Deflection of the cutting blade 30 is reliably suppressed. Although the degree of regulation of the movement of the cutting blade 30 is inferior to this case, the guide slit 31
May be formed only on one side of the accommodation space 11a.

According to the pipette according to the present embodiment, the same advantages as those of the pipette shown in FIG.
The following advantages are obtained. That is, when cutting or incising a cell or the like as the operation target 1 sucked and held by the suction port 10a, since the guide slit 31 is formed in the accommodation portion 11, the cutting blade 30 is inserted into the guide slit 31. Thereby, the cutting blade 30 can be guided by the guide slit 31 to cut the held cells and the like. As described above, not only the swing of the cells and the like is suppressed, but also the movement of the cutting blade 30 is substantially restricted to a predetermined plane by the guide slit 31, so that the cutting blade 30
Since blurring is also suppressed, cells or the like can be easily cut or incised along a desired plane. In the pipette shown in FIG. 1, since the cutting blade 30 cannot hit the edge of the housing 11 and enter the housing space 11 a, the operation target 1
In this case, only the portion protruding from the accommodation space 11a can be cut.

As the cutting blade 30, a thin plate is used, for example, a metal blade or the like may be used.
It may be a thin film plate or the like of a microdissector disclosed in Japanese Patent No. 5018. The micro-cutting device comprises a support for supporting a thin-film plate (cutting blade) and the thin-film plate protruding from the support and formed by using a semiconductor manufacturing technique. And the like, and cuts or cuts an operation target (processing target) while vibrating at a high frequency by a piezoelectric actuator as needed. In addition, a thin film reinforcing member for reinforcing the thin film plate is provided, a saw blade is formed on the thin film plate, both ends of the thin film plate are tapered as shown in FIG. The vicinity of the center of the thin film plate may have a waveform shape.

Next, an example of a method for manufacturing the pipette shown in FIG. 5 will be described with reference to FIGS. FIG. 6 is a cross-sectional view showing each step of the method for manufacturing the pipette shown in FIG. FIG. 7 is a cross-sectional view showing each step following the step shown in FIG. 6 in the method of manufacturing the pipette shown in FIG. The cross sections shown in FIGS. 6 and 7 correspond to YZ in FIG.
Corresponds to the cross section. FIG. 8 is a plan view showing a predetermined step shown in FIG. 6, and FIG. 8 (a) is DD ′ in FIG. 6 (a).
FIG. 8B is an arrow view, and FIG. 8B is an EE ′ arrow view in FIG. 9A and 9B are views showing the predetermined steps shown in FIGS. 6 and 7, wherein FIG. 9A is a view taken along the line FF ′ in FIG. 6D, and FIG. ), GG ′ arrow view, FIG.
FIG. 10 is a view taken along the line HH ′ in FIG. 6 to 9, the same or corresponding elements as those in FIGS. 2 to 4 are denoted by the same reference numerals.

First, a silicon substrate 20 having a diameter of 3 inches and a thickness of 400 μm and having a (100) plane orientation was used as a substrate material. Dichlorosilane and ammonia gas were used as mask materials on both surfaces of the substrate 20 by low-pressure vapor deposition. Then, silicon nitride films 21 and 22 are formed to a thickness of 100 nm. Further, by patterning the silicon nitride film 21 on the upper surface of the substrate 20 by a photolithography method and a dry etching method, a plurality of openings 21a for exposing the upper surface of the substrate 20 are formed at predetermined positions of the silicon nitride film 21 on the upper surface of the substrate 20. Form. The size of the opening 21 is, for example, 150 μm × 2
It is 50 μm. Thereafter, the substrate is immersed in an etching solution for silicon such as an aqueous solution of potassium hydroxide (KOH) or an aqueous solution of tetramethylammonium hydroxide (TMAH), and the substrate exposed from the opening 21a using the silicon nitride films 21 and 22 as masks. By etching the portion 20, a plurality of recesses 23 having a depth of 50 μm are simultaneously formed on the upper surface of the substrate 20. (FIGS. 6A and 8A). Since the substrate 20 has a (100) plane orientation, as is well known, etching is performed on silicon (1).
11) Since the surface of the recess 23 is automatically stopped at the surface, the surface of the recess 23 becomes a taper surface having a predetermined angle. In the present embodiment, one recess 23 includes a portion corresponding to two storage portions 11 corresponding to two pipettes.

Next, a recess having a depth of 10 μm including a portion corresponding to the thin tube portion 10 is formed in the same manner as the recess 23 is formed after the silicon nitride films 21 and 22 are once removed. In the embodiment, a plurality of grooves 24 are formed on the upper surface of the substrate 20.
The recess 23 and the corresponding recess 24 are continuous. Further, after the silicon nitride films (not shown) on the upper and lower surfaces formed at that time are once removed, a recess 40 having a depth of 1 μm including a portion corresponding to the guide slit 31 of the housing portion 11 is formed on the upper surface of the substrate 20. (FIG. 6B, FIG. 8)
(B)). The recess 23 and the corresponding recess 40 are continuous. In this case, the silicon nitride film remains on both surfaces of the substrate 20, but the silicon nitride film (not shown) on the upper surface is removed, leaving only the silicon nitride film 25 on the lower surface. FIG.
FIG. 8B and FIG. 8B show this state.

The recesses 23 and 40 including the portion corresponding to the housing 11 and the recess 2 including the portion corresponding to the thin tube portion 10 are provided.
4 may be formed by dry etching or the like without using wet etching, or may be formed by dicing (mechanical processing using a dicing saw or a processing machine similar thereto).

Two silicon substrates 20 in the state shown in FIG. 6B and FIG. 8B are prepared, and the recesses 23, 24, and 40 are provided inside the joined body of one substrate and the other substrate. The two substrates are joined together so that a cavity is formed (FIG. 6C). In the present embodiment, the surfaces of the two silicon substrates 20 where the recesses 23, 24, and 40 are formed are joined. This bonding is performed at a temperature of about 1200 ° C. by, for example, a direct bonding method.

Then, a groove 2 is formed in the joined body shown in FIG.
6 (FIGS. 6D and 9A). In the present embodiment, so as to reach the recess 23 from the upper surface of the joined body,
A groove 26 having a width of 30 μm is formed by dicing. As shown in FIG. 9A, the groove 26 is processed linearly, but is aligned so as to penetrate the plurality of recesses 23 by one processing. This groove 26
In the thermal oxidation treatment of the inner wall of the cavity described below,
It is formed so that oxygen is sufficiently supplied into the cavity. However, if the recesses 23, 24, and 40 are previously formed in the silicon substrate 20 so that the cavity communicates with the outside in the first place, and oxygen is sufficiently supplied into the cavity, it is not always necessary. , Need not be formed.

Next, the joined body in the state shown in FIG. 6D and FIG. 9A is placed in an electric furnace, and the inner wall (silicon portion) of the cavity formed by the recesses 23 and 24 is thermally oxidized (FIG. 7).
(A)). At this time, at the same time, the inner wall of the groove 26 and the recess 4
0 inner wall is also thermally oxidized. 2 in FIG.
Reference numeral 7 denotes a silicon oxide film formed by thermal oxidation. The thickness of the silicon oxide film 27 is, for example, about 2 μm.

Next, a groove having a width of 100 μm is used to separate the central portion of the recess 23 along the groove 26 by using a blade wider than the processing blade used for dicing for forming the groove 26. 28 (FIGS. 7B and 9).
(B)). As a result, the silicon oxide film 27 formed on the inner wall of one recess 23 is separated into ones corresponding to individual pipettes, and portions of the silicon oxide film 27 that do not correspond to the thin tube portion 10 and the housing portion 11. Then, the unnecessary portion formed on the inner wall of the groove 26 is removed.
Further, a groove 41 is formed in the bonded body by dicing.
Is formed in a direction perpendicular to the silicon oxide film 2.
The portion formed on the inner wall of the groove 40, which is a portion that does not correspond to the narrow tube portion 10 and the housing portion 11 of the silicon oxide film 7 and is unnecessary, is replaced with the thin tube portion 10 and the housing portion 1
The portion corresponding to 1 is made discontinuous (FIG. 9B)
(C)).

Thereafter, the silicon nitride films 25 on the upper and lower surfaces of the joined body are formed.
Is patterned by photolithography and dry etching in accordance with the desired shape of the support 12 (FIG. 7C).

Finally, the conjugate in the state shown in FIG. 7C was replaced with tetramethylammonium hydroxide (TMA).
H) Immerse in an aqueous solution (this aqueous solution hardly etches the silicon oxide film) and remove unnecessary silicon portions (support 2
Is removed by etching (FIG. 7D). Thereby, a plurality of pipettes shown in FIG. 5 are completed collectively. At this time, depending on the depth of the groove 41, the support 12
In some cases, the portions on both sides of the groove 41 may not be separated. In such a case, a force may be applied to the portion of the groove 41 to break it, thereby separating the two. However, even in such a case, it is not always necessary to break.

Further, since the pipette according to the present embodiment can be manufactured by using the semiconductor manufacturing technique as described above, it is possible to obtain the suction port 10a having an arbitrary shape and high accuracy, and It is also possible to manufacture a plurality of pipettes simultaneously and collectively, and it is possible to manufacture pipettes at a lower cost.

In the above-described manufacturing method, the bonding 2
Although a recess including a portion corresponding to the thin tube portion 10 and a portion corresponding to the housing portion 11 is formed on both of the two silicon substrates 20, in the present invention, the recess is formed only on one of the substrates. Is also good. Even in this case, a cavity having an inner wall of silicon can be formed in the joined body of the two substrates.

While the embodiments of the present invention have been described above, the present invention is not limited to these embodiments.

[0070]

As described above, according to the present invention,
It is possible to provide a pipette capable of reducing the deformation of the operation target due to the operation even if the operation target has good operability and the operation target is easily deformed.

Further, according to the present invention, it is possible to provide an inexpensive pipette having an arbitrary size and a high-precision suction port, easy to manufacture, and inexpensive, and a method for manufacturing the same.

Further, the present invention can provide a pipette capable of easily cutting or incising an operation target along a desired plane.

Further, according to the present invention, a plurality of high-precision micro-capillaries can be simultaneously manufactured at once.

[Brief description of the drawings]

FIG. 1 is a diagram schematically showing a pipette according to an embodiment of the present invention, and FIG. 1 (a) is a schematic perspective view thereof, FIG.
(B) is a schematic sectional view of a main part thereof.

FIG. 2 is a cross-sectional view showing each step of the method for manufacturing the pipette shown in FIG.

FIG. 3 is a cross-sectional view showing each step of the method of manufacturing the pipette shown in FIG. 1 subsequent to the step shown in FIG. 2;

FIG. 4 is a plan view showing a predetermined step shown in FIG. 2;

FIG. 5 is a view schematically showing a pipette according to another embodiment of the present invention, and FIG. 5 (a) is a schematic perspective view thereof, FIG.
(B) is a schematic sectional view of a main part thereof.

6 is a cross-sectional view showing each step of the method for manufacturing the pipette shown in FIG.

FIG. 7 is a cross-sectional view showing each step following the step shown in FIG. 6 of the method of manufacturing the pipette shown in FIG.

FIG. 8 is a plan view showing a predetermined step shown in FIG.

FIG. 9 is a view showing a predetermined step shown in FIGS. 6 and 7;

FIG. 10 is a cross-sectional view showing a main part of a conventional pipette.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Operation target 5 Sperm injection glass tube 10 Thin tube part 10a Suction port 11 Storage part 11a Storage space 12 Support body 12a Connection opening 12b Internal passage 20 Silicon substrate 23,24,40 Recess 26,28,41 Groove 27 Silicon oxide film 30 cutting blade 31 guide slit

Claims (9)

[Claims] A thin tube portion having an opening at a distal end serving as a suction port; and a thin tube portion integrally formed at the distal end of the thin tube portion,
A cup-shaped housing portion that surrounds the periphery of the operation target sucked and held by the suction port from a side in the suction direction of the operation target by the suction port to form a storage space for the operation target. Features pipette. 2. The pipette according to claim 1, wherein said thin tube portion and said housing portion are integrally formed of silicon oxide. 3. A support for supporting a proximal end of the thin tube portion, the connection opening being open to the outside and drawing a negative pressure, and communicating between the inside of the thin tube portion and the connection port. The pipette according to claim 1, further comprising a support having an internal passage. 4. A guide slit, which guides a cutting blade for cutting the operation target into the housing space and regulates the movement of the cutting blade substantially within a predetermined plane, is formed in the housing portion. The pipette according to any one of claims 1 to 3, wherein 5. The storage device according to claim 4, wherein the guide slits are formed at both sides of the housing space in the housing so that the cutting blades can be inserted simultaneously across the housing space. Pipette. 6. A pipette manufacturing method for manufacturing the pipette according to claim 1, wherein the first and second silicon substrates are provided.
Preparing a first and second silicon substrate in which a recess is formed on both or one surface of the silicon substrate, the recess including at least a portion corresponding to the capillary portion and a portion corresponding to the housing portion; And forming the first silicon substrate and the second silicon substrate so that a cavity is formed by the recess inside the joined body of the first silicon substrate and the second silicon substrate. Bonding, a step of thermally oxidizing a wall surface of the cavity, and a step of etching at least a part of a portion of the first and second silicon substrates that is not thermally oxidized. A method for producing a pipette. 7. The method according to claim 1, further comprising, after the joining step and before the thermal oxidation step, a step of forming a groove in the joined body for communicating the cavity with the outside. 7. The method for producing a pipette according to 6. 8. The method according to claim 1, further comprising the step of:
And removing at least a part of the portion of the thermally oxidized portion of the second silicon substrate that does not correspond to the thin tube portion and the housing portion, or removing the thermally oxidized portion of the first and second silicon substrates. The pipette manufacturing method according to claim 6, further comprising the step of discontinuing a portion corresponding to the thin tube portion and the storage portion of the portion that is formed. 9. A method for manufacturing a thin tube, comprising: first and second silicon substrates, wherein the first and second silicon substrates are provided.
Providing a first and a second silicon substrate having a recess formed on both or one surface of the silicon substrate, the recess including at least a portion corresponding to the capillary; and Bonding the first silicon substrate and the second silicon substrate such that a cavity is formed by the recess inside a bonded body of the silicon substrate and the second silicon substrate; Producing a thin tube, comprising: thermally oxidizing a wall surface of a cavity; and removing at least a part of a portion of the first and second silicon substrates that is not thermally oxidized by etching. Method.