JPH09108578A - Pipette and its manufacture - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a pipette which has a suction port of any size and of high accuracy and can be easily manufactured at a low cost. SOLUTION: A pipette body 10 has a suction port 11 which is open to the exterior to hold an object to be treated by suction, a connection port 12 which is open to the exterior for sucking by vacuum suction, and an interior passage 13 for interconnecting the ports 11, 12. The body 10 comprises a connected body in which a first member 14 is connected to a second member 15. In at least one of the joint surfaces of the members 14, 15, there is formed a recess 13a corresponding to at least a portion of the passage 13.

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, glass pipettes have been used to manipulate microscopic objects such as cells. FIG. 8 is a sectional view showing a main part of this conventional pipette. As shown in FIG. 8, this conventional pipette has a suction port 2 which is opened to the outside to suck and hold an operation target 1 such as a cell, and a connection port (not shown) which is opened to the outside to draw a negative pressure. ), And a pipette body 4 made of a glass tube having a suction port 2 and an internal passage 3 communicating with the connection port. The connection port is an opening of the internal passage 3 on the opposite side of the suction port 2 in the glass tube which is the pipette body 4.

According to this pipette, the connection port side portion of the pipette body 4 is connected to, for example, a so-called injector (which is configured like a syringe), and the injector is operated to form an internal passage through the connection port. Pull the inside of 3 to negative pressure. As a result, as shown in FIG.
Are suction-held in the suction port 2. In this state, for example, when the operation target 1 is an egg, a needle (not shown) is inserted into the egg to inject sperm to perform fertilization, or the operation target 1 is various cells or the like. In this case, various operations are performed such as puncturing the operation target 1 with a needle or the like to open a hole. In addition, after completion of such an operation, 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). As a result, the pipe becomes thin and cut, and two pipette bodies 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 when it is cut, the operation target 1 will be damaged if it is used as it is. Therefore, the tip of the pipette is heated by a device (not shown) such as a microforge to remove the edge. Round it.

[0005]

However, since the conventional pipette is manufactured by the manufacturing method described above, the method of drawing and heating the glass tube depends on personal experience in manufacturing the conventional pipette. The shape of the manufactured pipette differs depending on the type, and a skilled technique is required to manufacture the pipette having 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 an object of the present invention is to provide a pipette having an arbitrary size and a highly accurate suction port, which is easy and inexpensive to manufacture, and a manufacturing method thereof. .

[0007]

In order to solve the above-mentioned problems, a pipette according to a first aspect of the present invention has a suction port which is opened to the outside to suck and hold an operation target, and an outside which is opened to a negative pressure. The pipette main body has a connection port for pulling and an internal passage that connects the suction port and the connection port. And, the pipette body is
And a second member having a second flat surface, wherein the first flat surface and the second flat surface are joined together. At least one of the first plane of the first member and the second plane of the second member is provided with a recess corresponding to at least a part of the internal passage. is there.

Since the pipette according to the first aspect has such a structure, one of the first plane of the first member and the second plane of the second member or A recess corresponding to at least a part of the internal passage is formed in both of them, and the first plane of the first member and the second plane of the second member are joined to each other to form a pipette. It can be manufactured. Therefore, for example, the recess can be formed by using a semiconductor manufacturing technique such as etching or dicing.
The pipette according to the first aspect has a suction port having an arbitrary shape and high precision, and is easy to manufacture and inexpensive.

In the pipette according to the first aspect, the cross-sectional area of the internal passage may be the same throughout,
In that case, if the operation target is extremely small, the suction port is made small accordingly, so that the cross-sectional area of the internal passage is reduced as a whole. For this reason, the conductance of the internal passage with respect to the flow of the fluid becomes small, and a large force is required to pull the internal passage to a negative pressure via the connection port by the injector or the like, which is not preferable.

Therefore, in the pipette according to the second aspect of the present invention, in the pipette according to the first aspect, as compared with the cross-sectional area of the portion near the suction port in the internal passage, It has a large cross-sectional area. The pipette according to the third aspect of the present invention also comprises
In the pipette according to the first aspect, the cross-sectional area of the internal passage is increased from the suction port side toward the connection port side.

According to the pipettes according to the second and third aspects, by constructing the internal passage as described above, even if the object to be operated is extremely small and the suction port is made small accordingly. Since the conductance of the internal passage with respect to the flow of the fluid becomes large, a large force is not required when pulling the internal passage to a negative pressure via the connection port by an injector or the like, which is preferable.

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 at least one of the first and second members is made of a transparent material. is there. In the first to third aspects, the material of the first and second members may be an opaque material, but according to the fourth aspect, the operation of the suction port and the state of being suction-held in the suction port. The target is reflected light when only one of the first and second members is made of a transparent material, and when both of the first and second members are made of a transparent material. The reflected light and the transmitted light allow observation and facilitate the operation of the operation target.

A pipette according to a fifth aspect of the present invention is the pipette according to any one of the first to third aspects, wherein at least one of the first and second members is made of silicon. In the first to third aspects, the material of the first and second members is not particularly limited, but according to the fifth aspect, the first and second
Since at least one of the members is made of silicon, the member can be formed by using an inexpensive silicon wafer whose surface is processed flat and smooth, and etching or etching is not performed on the silicon wafer. It is preferable because dicing is easy.

A pipette according to a sixth aspect of the present invention is the pipette according to any one of the first to third aspects, wherein the first member is made of a glass material containing mobile ions, and the second member is It is made of semiconductor or metal. In the first to third aspects, the first member and the second
For joining with the members of No. 3, any joining method such as joining with an adhesive can be adopted. In the sixth aspect, the materials as described above are used as the first and second members. Therefore, the first member and the second member can be bonded by anodic bonding, and by adopting the anodic bonding in this way, the first member and the second member can be bonded. Joining becomes easy and strong and stable joining becomes possible. In addition, since the first member is made of a glass member, and thus is made of a transparent member, it is possible to observe the suction port and the operation target in a state of being suction-held by the suction port, and the operation of the operation target is easy. become.

A pipette manufacturing method according to a seventh aspect of the present invention is a pipette manufacturing method for manufacturing the pipette according to any one of the first to third aspects, which comprises:
Of the first and second substrates, wherein a recess corresponding to at least a part of the internal passage for a plurality of pipettes is formed on one plane of at least one of the first and second substrates. Providing a second substrate, joining between the plane of the first substrate and the plane of the second substrate,
Separating the bonded first and second substrates into individual pipettes. According to the seventh aspect, since a plurality of pipettes can be manufactured at one time, the manufacturing thereof becomes easier and more inexpensive.

The pipette manufacturing method according to an eighth aspect of the present invention is the pipette manufacturing method according to the seventh aspect, wherein the step of preparing the first and second substrates is performed by etching or dicing to obtain the pipette. 1st and 2nd
Forming a recess in one plane of at least one of the substrates. The dicing process means a mechanical process using a dicing saw or a processing machine similar to this. According to the eighth aspect, since the etching or dicing process adopted in the semiconductor manufacturing technique is used for forming the recess, it is possible to realize a suction port or the like having an arbitrary size and high accuracy. it can.

A pipette manufacturing method according to a ninth aspect of the present invention is the pipette manufacturing method according to the seventh or eighth aspect, wherein the first substrate is made of a glass material containing mobile ions, and the second substrate is Is made of a semiconductor or a metal, and the bonding step includes a step of anodic bonding between the plane of the first substrate and the plane of the second substrate. In the ninth aspect, since anodic bonding is adopted as the bonding between the first substrate and the second substrate, the bonding between them is facilitated and the strong and stable bonding is possible.

[0018]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Various embodiments of the present invention will be described in detail below with reference to the drawings.

First, a pipette according to a first embodiment of the present invention will be described with reference to FIG.

FIG. 1 (a) is a perspective view showing a pipette according to this embodiment, and FIG. 1 (b) is a plan view of the second member 15 constituting the pipette according to this embodiment as seen from the upper surface side. . In order to facilitate understanding of the drawing, FIG.
In FIG. 5, a line that is not actually visible and should be a hidden line (broken line) is also shown by a solid line (see FIGS.
The same applies to (a) and FIG. 7 (a). ).

The pipette according to the present embodiment has a suction port 11 which is opened to the outside to suck and hold an operation target (not shown).
And a connection port 12 for opening to the outside to draw a negative pressure, and an internal passage 13 for communicating between the suction port 11 and the connection port 12.
And a pipette body 10 having And
The pipette body 10 includes a first member 14 having a first plane (lower surface in this example) and a second plane (upper surface in this example).
And a second member 15 having a structure, which is formed by joining the lower surface of the first member 14 and the upper surface of the second member 15 to each other. In addition, in this Embodiment, although the 1st and 2nd members 14 and 15 are comprised by flat plate shape, the shape of the upper surface of the 1st member 14 and the lower surface of the 2nd member 15 is not necessarily planar. It is not something that will be done.

In the present embodiment, the upper surface of the second member 15 is provided with a recess (a linearly extending groove) 13a corresponding to the entire internal passage 13. The recess 1
One end 12a of 3a corresponds to the connection port 12, and the other end 11a of the recess 13a corresponds to the suction port 11.
On the other hand, no recess is formed on the lower surface of the first member 14, and the lower surface of the first member 14 is simply a flat surface. However, not only the upper surface of the second member 15 but also the lower surface of the first member 14 is formed with a recess, so that the first member 1
The internal passage 13 may be constituted by a recess on the lower surface of 4 and a recess on the upper surface of the second member 15.

Next, an example of a method of manufacturing the above-mentioned pipette shown in FIG. 1 will be described with reference to FIG. FIG.
FIG. 3 is a cross-sectional view showing each step of the method for manufacturing the pipette shown in FIG. 1.

First, as shown in FIG. 2A, the second
A silicon substrate 20 to be the member 15 is prepared.

Next, as shown in FIG. 2B, the recesses 13a corresponding to a plurality of pipettes are formed on the upper surface of the silicon substrate (silicon wafer) 20. In addition, in FIG. 2A, although the recesses 13a for three pipettes are shown,
The number is not limited. The recess 13a can be formed by using, for example, semiconductor manufacturing technology such as etching and dicing. In FIG. 3A, the recess 13 is formed on the upper surface of the silicon substrate 20 by dicing.
An example in which a is formed is shown. In this case, the size of the recess 13a can be determined by the blade width of the dicing saw used during the dicing process and the amount of cut into the substrate 20. Further, FIG. 3B shows an example in which the recess 13a is formed on the upper surface of the silicon substrate 20 by isotropic etching. In this case, nitric acid + hydrofluoric acid can be used as the etching liquid. Furthermore, FIG.
Shows an example in which the recess 13a is formed on the upper surface of the silicon substrate 20 by anisotropic etching. In this case, for example, a silicon single crystal substrate having a (100) plane orientation is used as the silicon substrate 20, and KOH is used as an etching solution.
A solution can be used. At this time, the sidewall of the recess 13a forms an angle of 54.7 ° with the surface of the substrate 20 (11
1) It becomes the aspect of the system. When etching is adopted as shown in FIGS. 3B and 3C, the conditions of the concentration of the etching solution, the temperature, the etching time, and the mask shape (not shown) at the time of etching are set to a desired shape and Dimensional recess 1
It may be set appropriately so as to obtain 3a. By adopting dicing processing and etching as described above,
It is possible to form the recess 13a having various cross-sectional shapes and dimensions with an accuracy of a micron unit. 1 and 2 show an example in which the recess 13a is formed by dicing as shown in FIG. 3 (a).

After that, as shown in FIG. 2C, a lower surface of a glass substrate 21 made of a glass material (for example, borosilicate glass) containing mobile ions to serve as the first member 14 prepared in advance, The upper surface of the silicon substrate 20 in which the recess 13a is formed is bonded. This bonding can be performed by, for example, anodic bonding. FIG. 4 schematically shows the manner of anodic bonding between the silicon substrate 20 and the glass substrate 21. In this anodic bonding, for example, the silicon substrate 20 and the glass substrate 21 are heated to 400 ° C. by the hot plate 30, and a DC voltage of 600 V is applied to the silicon substrate 20,
After maintaining in that state for 10 minutes, the applied voltage is lowered to 0 V, the heating of the hot plate 30 is stopped, and the hot plate 30 is naturally cooled. However, the silicon substrate 20
The bonding between the glass substrate 21 and the glass substrate 21 is not limited to anodic bonding, and they may be bonded using an adhesive or the like.

Finally, as shown in FIG. 2D, the bonded silicon substrate 20 and glass substrate 21 are separated into individual pipettes by using a dicing saw or the like.
It is possible to obtain a plurality of pipettes shown in FIG. 2 at one time.

In the manufacturing method described above, the first member 1
A glass substrate 21 made of a glass material containing mobile ions is used as the first substrate to be No. 4, and the second member 1
Since the silicon substrate 20 is used as the second substrate which should be 5, the first member 14 is made of a glass material containing mobile ions, and the second member 15 is made of silicon. The material of the second members 14 and 15 is not limited to these, and any material can be used. However, as the material of the member forming the recess, a semiconductor, a metal, a glass material or the like is selected when the formation is performed by etching. When the recess is formed by dicing, the material of the member forming the recess is not limited. Silicon is an indispensable material for semiconductor technology, and the surface of a silicon wafer is processed to be flat and smooth.
Alternatively, it is a material suitable for the second substrate. First and second
If at least one of the members 14 and 15 is a transparent material such as a glass material, the suction port 11 and the operation target sucked and held by the suction port 11 can be observed by reflected light or transmitted light. This is preferable because it facilitates the operation. In this case, if the anodic bonding as described above is not adopted, a transparent material such as a glass material containing no mobile ions may be adopted.

According to the pipette shown in FIG. 1 according to the present embodiment, the portion of the pipette body 10 on the side of the connection port 12 is connected to, for example, an injector, and the injector is operated to move the inside of the internal passage 13 through the connection port 12. Pull to negative pressure. As a result, the operation target such as cells is suction-held in the suction port 11.
Then, in this state, for example, when the operation target is an egg, insert a needle into the egg to inject sperm to perform fertilization, or when the operation target is various cells etc., the operation target is a needle, etc. Performs various operations, such as puncturing to make a hole. Note that after such operations are completed,
The inside of the internal passage 13 is made to have a positive pressure through the connection port 12 by operating the injector, and the suction holding state of the operation target by the suction port 11 is released.

Then, the pipette according to the present embodiment is
Since it can be manufactured as described above, the suction port 11 having an arbitrary shape and high accuracy can be obtained, and the manufacturing is easy and inexpensive.

Next, a pipette according to a second embodiment of the present invention will be described with reference to FIG.

FIG. 5A is a perspective view showing the pipette according to this embodiment, and FIG. 5B is a plan view seen from the upper surface side of the second member 15 constituting the pipette according to this embodiment. . 5, elements that are the same as or correspond to the elements in FIG. 1 are assigned the same reference numerals and explanations thereof are omitted.

The pipette according to this embodiment is the same as the pipette shown in FIG.
And two sets of internal passages 13 are provided. The pipette according to the present embodiment has, for example, a suction port 11, a connection port 12 and an internal passage 13 when being separated into individual pipettes after undergoing the steps shown in FIGS. 2A to 2C described above.
Can be manufactured by separating so as to have two sets. The suction port 11 and the connection port 12 of each set may be different from each other in shape and size.

According to the present embodiment, it is possible to hold a plurality of operation targets by one sampling, which enables efficient operation.

The suction port 11, the connection port 12 and the internal passage 13 may be provided in three or more sets. Also, the connection port 12 is 1
It is also possible to form only the individual pieces and to have the internal passage 13 extend from the connection port 12 and branch into the plurality of suction ports 11.

Next, a pipette according to a third embodiment of the present invention will be described with reference to FIG.

FIG. 6A is a perspective view showing the pipette according to this embodiment, and FIG. 6B is a plan view seen from the upper surface side of the second member 15 constituting the pipette according to this embodiment. . 6, elements that are the same as or correspond to the elements in FIG. 1 are assigned the same reference numerals, and descriptions thereof will be omitted.

In the present embodiment, a recess 40a having a large cross-sectional area corresponding to a part of the internal passage 13 and a recess having a small cross-sectional area corresponding to the rest of the internal passage 13 are formed on the upper surface of the second member 15. 40b is formed. One end 41 of the recess 40a
a corresponds to the connection port 12, and the other end of the recess 40a is the recess 40
b, one end of which is continuous with the other end 42a of the recess 40b, and the suction port 1
It corresponds to 1.

Therefore, in the present embodiment, compared to the cross-sectional area of the portion of the internal passage 13 on the suction port 11 side (the portion corresponding to the recess 40b), the other portion of the internal passage 13 (corresponding to the recess 40a). The cross-sectional area of the part to

The pipette according to this embodiment can be manufactured in the same manner as the pipette shown in FIG. 1, and the same advantages as the pipette shown in FIG. 2 can be obtained. Further, according to the present embodiment, as described above, since the cross-sectional area of the other portion of the internal passage 13 is larger than the cross-sectional area of the portion of the internal passage 13 on the suction port 11 side, the operation target is extremely large. Even if the suction port 11 is made small and accordingly small, the conductance of the internal passage 13 with respect to the flow of the fluid becomes large. Therefore, a large force is not required when the internal passage 13 is pulled to a negative pressure via the connection port 12 by an injector or the like, which is preferable.

Next, a pipette according to a fourth embodiment of the present invention will be described with reference to FIG.

FIG. 7A is a perspective view showing the pipette according to this embodiment, and FIG. 7B is a plan view seen from the upper surface side of the second member 15 constituting the pipette according to this embodiment. . 7, elements that are the same as or correspond to the elements in FIG. 1 are assigned the same reference numerals and explanations thereof are omitted.

In the present embodiment, the upper surface of the second member 15 is provided with a recess 50a corresponding to the entire internal passage 13. One end 52a of the recess 50a corresponds to the connection port 12, and the other end 51a of the recess 50a corresponds to the suction port 11. The cross-sectional area of the recess 50a increases from the end 51a toward the end 52a. Such a recess 50a is formed by using, for example, the second member 15 using a silicon single crystal substrate having a (100) plane orientation, using a mask having a tapered pattern, and using a KOH solution as an etching solution. It can be obtained by subjecting the substrate to the anisotropic etching. In this case, the recess 50a has a triangular cross-sectional shape as shown in FIG.

Therefore, in the present embodiment, the cross-sectional area of the internal passage 13 increases from the suction port 11 side toward the connection port 12 side.

The pipette according to this embodiment can be manufactured in the same manner as the pipette shown in FIG. 1, and the same advantages as the pipette shown in FIG. 2 can be obtained. Furthermore, according to the present embodiment, as described above, since the cross-sectional area of the internal passage 13 is larger from the suction port 11 side toward the connection port 12 side,
Even when the operation target is extremely small and the suction port 11 is configured to be small accordingly, the conductance of the internal passage 13 with respect to the flow of the fluid becomes large. Therefore, a large force is not required when the internal passage 13 is pulled to a negative pressure via the connection port 12 by an injector or the like, which is preferable.

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

[0047]

As described above, according to the present invention,
It is possible to provide a pipette having an arbitrary size and a highly accurate suction port, easy to manufacture and inexpensive, and a manufacturing method thereof.

[Brief description of the drawings]

FIG. 1 shows a pipette according to a first embodiment of the present invention.

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

FIG. 3 is a cross-sectional view showing a method of forming a recess.

FIG. 4 is a diagram schematically showing the manner of anodic bonding between a silicon substrate and a glass substrate.

FIG. 5 shows a pipette according to a second embodiment of the invention.

FIG. 6 shows a pipette according to a third embodiment of the invention.

FIG. 7 shows a pipette according to a fourth embodiment of the invention.

FIG. 8 is a sectional view showing a main part of a conventional pipette.

[Explanation of symbols]

 10 Pipette Body 11 Suction Port 12 Connection Port 13 Internal Passage 13a, 40a, 40b, 50a Recess 14 First Member 15 Second Member 20 Silicon Substrate 21 Glass Substrate 30 Hot Plate

Claims (9)

[Claims] 1. A suction port that is opened to the outside to hold an operation target by suction, and a connection port that is opened to the outside to draw a negative pressure.
A pipette body having an internal passage communicating between the suction port and the connection port, wherein the pipette body has a first member having a first plane and a second member having a second plane. A joined body consisting of a joined body in which the first plane and the second plane are joined together, wherein the first plane of the first member and the second member A pipette, wherein a recess corresponding to at least a part of the internal passage is formed on at least one of the second planes. 2. The pipette according to claim 1, wherein a cross-sectional area of another portion of the internal passage is larger than a cross-sectional area of a portion of the internal passage near the suction port. 3. The pipette according to claim 1, wherein the cross-sectional area of the internal passage is larger from the suction port side toward the connection port side. 4. The at least one of the first and second members is made of a transparent material.
The pipette according to any one of 1 to 3. 5. The at least one of the first and second members is made of silicon.
The pipette according to any one of 1 to 3. 6. The pipette according to claim 1, wherein the first member is made of a glass material containing mobile ions, and the second member is made of a semiconductor or a metal. 7. A pipette manufacturing method for manufacturing the pipette according to claim 1, wherein the first and second substrates are at least one of the first and second substrates. Providing a first and a second substrate in which a recess corresponding to at least a part of the internal passage for a plurality of pipettes is formed in one plane of the substrate; and a plane of the first substrate. A method of manufacturing a pipette, comprising: a step of joining between the flat surfaces of the second substrate; and a step of separating the joined first and second substrates into individual pipettes. 8. The step of preparing the first and second substrates forms the recess in one plane of at least one of the first and second substrates by etching or dicing. The pipette manufacturing method according to claim 7, further comprising steps. 9. The first substrate is made of a glass material containing mobile ions, the second substrate is made of a semiconductor or a metal, and the bonding step is performed on the plane of the first substrate and the second substrate. 9. The pipette manufacturing method according to claim 7, further comprising a step of anodic bonding with the plane of the substrate.