JP2002228671A - Suction and discharge-switching device for sample solution spotting apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a suction and discharge-switching device for a sample solution spotting device for easily switching a number of channels for connecting a suction member and a dispensing member with a suction and discharge operation member when spotting a sample probe onto a substrate and nearly uniform quantity, for enabling efficient spotting operation to be performed. SOLUTION: A sample solution is spotted onto a substrate in a matrix to manufacture a sample chip. A switching member having a number of suction channels connected to each suction member for sucking sample solution being accumulated at each well section of a container body and a number of discharge channels for connecting the sucked sample solution to each dispensing member is moved to a number of suction discharge channels that suck and accumulate the sample solution being accumulated at each well section of the container body and are connected to each suction discharge operation member for determining and discharging the sample solution to each dispensing member, thus selectively matching each suction channel and discharge channel corresponding to each suction discharge channel for connection.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a DNA,
A sample solution is suctioned from a container in which a large number of various sample solutions are stored in a sample solution spotting apparatus for preparing a large number of sample arrays of the same type in which a large number of sample probes such as RNA polynucleotides and proteins are fixed in spots. The present invention relates to a suction / discharge switching device that switches a flow path and a flow path that discharges to a dispensing member.

[0002]

For example, research for screening and quantitative measurement of proteins and nucleic acids is carried out in a microtiter plate (80 mm × 120 mm, 96-well or 384-well).
The measurement is performed by injecting the sample solution into each hole of
In order to increase the number of samples to be measured at one time and to reduce the required sample volume, four 96-well microtiter plates were
Dispensing into one 4-well microtiter plate has been performed.

By performing this transfer operation using an automated machine, the amount of sample used is 10
In addition, the number of samples to be performed at one time can be reduced to one-fourth, and the number of samples to be performed at one time can be performed four times in 96 holes at a time.

However, the volume occupied by the microtiter plate remains unchanged at 80 mm × 120 mm, and at the present time when analysis is required with a very small amount of sample, several tens of microliters are required. Analytical systems are no longer in line. In addition, the number of analysis counts that can be performed at one time, as represented by a DNA microarray, is insufficient at 384.

[0005] Further, when automation is performed by a machine, the operation requiring human labor is simplified, and the suction site for sampling the sample is washed and re-used many times to reduce the occupied volume and cost of the entire apparatus. Often used, contamination between samples was inevitable.

SUMMARY OF THE INVENTION The present invention has been made in order to solve the above-mentioned conventional drawbacks. The object of the present invention is to solve such a problem on a small occupied substrate such as a slide glass (26 mm × 76 mm). A sample that spots a large number of samples at high density, and at that time, easily switches a large number of flow paths connecting a suction member and a dispensing member to a suction / discharge operation member, thereby enabling efficient spotting operation. An object of the present invention is to provide a suction / discharge switching device for a solution dispensing device.

[0007]

According to the present invention, there is provided a sample solution spotting apparatus for spotting a sample solution in a matrix on a substrate by using a plurality of dispensing members.
For a number of suction / discharge channels communicating with respective suction / discharge actuating members for sucking and storing the sample solution stored in each of the reservoir portions of the container body and for quantitatively discharging the stored sample solution to each dispensing member, Switching having a number of suction channels communicating with respective suction members for sucking the sample solution stored in the respective reservoirs of the container body and a number of discharge channels communicating with the respective dispensing members the suctioned sample solution. It is characterized in that the member is moved so that the respective suction flow paths and discharge flow paths corresponding to the respective suction / discharge flow paths are selectively matched to be able to communicate with each other.

[0008]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be described below with reference to the drawings. Embodiment 1 In FIGS. 1 to 4, a spotting device 1 includes a suction / discharge device 3 and a dispensing device 5.

A suction / discharge device 3 is disposed on the right side of the body frame 7 of the spotting device 1 in the drawing, and a first traveling body 9 of the suction / discharge device 3 is supported so as to be able to reciprocate in the front-rear direction as shown. The first traveling body 9 is provided with a first upper and lower frame 11, and the first upper and lower frame 11
The holder 13 is supported movably in the vertical direction.

As the driving device for reciprocating the first traveling body 9 with respect to the main body frame 7 and the first holder 13 with respect to the first upper and lower frames 11, respectively, the main body frame 7 and the first
A belt in which a part of a timing belt stretched on a pair of pulleys rotatably supported by the upper and lower frames 11 and one of which is connected to an electric motor such as a step motor is fixed to the first traveling body 9 and the first holder 13. A drive mechanism, a feed screw rotatably supported by the main body frame 7 and the first upper and lower frames 11 and having a shaft end to which an electric motor such as a step motor is connected are connected to a nut portion of the first traveling body 9 and the first holder 13. Any of a feed screw driving mechanism, a linear motor (neither is shown), or the like, which is meshed with the motor may be used.

The first holder 13 has a suction / discharge switching device 14
And a switching plate 17 that moves in close contact with the fixed plate 15. The fixed plate 15 has a large number (the number of sample probes spotted on the substrate 53) forming suction and discharge channels. Are 96) (96 in the case where the number of suction ports is 96). Further, the switching plate 17 has a large number (9
(Six) suction ports 19 are formed at intervals corresponding to the intervals between the suction / ejection ports 27, and are arranged adjacent to each suction port 19 to form a large number of ejection ports 21 forming an ejection channel. Are formed at predetermined intervals, and the suction port 19 and the discharge port 21 are set so as to selectively communicate with the corresponding suction / discharge port 27.

An air cylinder or a feed screw mechanism (not shown) is used as a drive mechanism for reciprocating the switching plate 17 at a distance corresponding to the distance between the suction port 19 and the discharge port 21.
Should be fine. In the above description, the movement of the switching plate 17 is controlled so that the suction port 19 is
And the discharge ports 21 are selectively matched to communicate with each other. However, the configuration may be such that the fixed plate 15 is moved and controlled to match.

Each of the suction ports 19 is provided with a suction needle 23 as a suction member having a vertical axis. The number of sample probes spotted on the substrate 53 is not limited to the above.

The tip of each suction needle 23 is opposed to a container 25 mounted on the main body frame 7. The container 25 is provided with, for example, the above-mentioned 96 reservoirs 25a facing the respective suction needles 23. Each reservoir 25a is provided with a predetermined amount (for example, 100 μl) of a different type or the same type of sample solution to be spotted. ) Each is stored separately.

The sample solution is adjusted according to the sample chip to be prepared, and DNA, RNA (DNA derived from chromosome)
Fragments, mRNA, cDNA, synthetic DNA and RNA, etc.)
For example, in a 3 × 3 SSC solution (adjusted to pH 7.00 with 0.45 M sodium chloride and 0.045 M sodium citrate).

A suction / discharge operation member 29 provided on the main frame 7 is connected to each suction / discharge port 27 of the fixed plate 15. The suction / discharge actuating member 29 is composed of a large number (96 in the above case) of injectors slidably supporting a piston 29b in a cylinder 29a, and each cylinder 29a has a corresponding suction / discharge port 27.
Is connected by a tube 33.

Each suction / discharge operating member 29 reciprocates the operating plate 31 connected to each piston 29b to suck the sample solution into the cylinder 29a including the tube 33 and discharge the stored sample solution. It is configured as follows. The operation plate 31 is configured to be minutely movable by, for example, a feed screw (neither is shown) connected to a step motor.

More specifically, the operating plate 31 is moved at a predetermined distance so that the entire amount of the sample solution stored in the pool 25a is sucked, and the sample solution in the pool 25a is transferred into the cylinder 29a including the tube 33. The operating plate 31 is minutely moved so that the amount of one discharge is, for example, 10 μl, and the sample solution in the cylinder 29 a including the tube 33 is discharged.

The dispensing device 5 described above is disposed on the left side of the main frame 7 as shown in the figure. The dispensing device 5 is connected to each discharge port 21 of the suction / discharge device 3 via a tube 35.
The dispensing device 5 is supported on the main body frame 7 by a second traveling body 37 movably in the left-right direction shown in the figure, and a third traveling body 37 is supported on the second traveling body 37 by a movably supported in the front-rear direction. A second movable body 39 supported by a traveling body 39 and a second upper and lower frame 41 provided on the third traveling body 39 so as to be vertically movable.
It comprises a holder 43 and a number (96 in the above case) of dispensing needles 47 provided on the second holder 43.

The second traveling body 37 with respect to the main body frame 7,
The driving mechanism for moving the third holder 39 with respect to the second running body 37 and the second holder 43 with respect to the second upper and lower frames 41, respectively, includes the above-described belt driving mechanism, feed screw driving mechanism, and linear motor. Any one may be used.

Each dispensing needle 47 has a tip surface diameter of 100 to 2
It is connected to each of the above-mentioned discharge ports 21 by respective tubes 35 connected to the base end portion at 00 μm. Each dispensing needle 47 is moved about 100 to 1 in the left-right direction and the front-rear direction shown in the drawing so that all the dispensing needles 47 are located in the substrate 53.
They are arranged in a matrix at intervals of 000 μm.

The interval between the dispensing needles 47 is determined from the relationship between the spotting area on the substrate 53 and the number of spotting, and is not limited to the above interval.

A substrate mounting table 51 is provided on the main body frame 7 below the dispensing needle group 49 in which a large number of dispensing needles 47 are arranged in a matrix. The substrate mounting table 51 has a slide glass and a film. A substrate 53 made of plastics (polyethylene, polypropylene) or the like is disposed so as to be positionable.
A preferred example of the substrate 53 is a slide glass. Preferably, an ultra-thin silicon rubber sheet having a large number of holes formed on the surface of the substrate 53 at intervals corresponding to the arrangement intervals of the dispensing needles 47 or the like. It is suitable to coat a thin film (not shown) such as an ultra-thin butyl rubber sheet and store the sample solution spotted in each hole of the silicon rubber sheet to prevent cross-contamination between spots.

A method for spotting a sample solution using the spotting apparatus 1 configured as described above will be described.

FIGS. 5 and 6 are explanatory views showing the switching state of the flow path. First, the movement of the first traveling body 9 of the suction / discharge device 3 is controlled so that each suction needle 23 is opposed to each of the pool portions 25a of the container 25 in which a predetermined sample solution is stored. Also,
The movement of the second traveling body 37 and the third traveling body 39 of the dispensing device 5 is controlled, and, for example, ten substrates 5 stacked in a multi-stage manner with the tip of each dispensing needle 47 positioned on the substrate mounting table 51.
3 relative to the spotting area. In the case of the substrate 53 whose surface is covered with a silicon rubber sheet, the tip of each dispensing needle 47 is opposed to each hole formed in the silicon rubber sheet.

The switching plate 17 of the suction / discharge switching device 14
Is moved such that each suction / ejection port 27 coincides with the corresponding suction port 19.

In the above state, the lowering of the first upper and lower frames 11 is controlled so that the tips of the suction needles 23 correspond to the corresponding reservoirs 25a.
After that, the operation plate 31 is operated to move the piston 29b of each suction / discharge operation member 29 in the suction direction, and the total amount of the sample solution stored in the storage portion 25a in the cylinder 29a including the tube 33 is stored. Aspirate.

Next, the switching plate 17 is moved so that each suction / discharge port 27 coincides with the corresponding discharge port 21, and the second upper / lower frame 41 of the dispensing device 5 is controlled to descend so that each dispensing needle 47 is moved. At a very small distance from the surface of the substrate 53.

In this state, the operating plate 31 is dispensed (for example, 1
0 μl) and control the movement at a distance corresponding to each tube 3
A large number of the sample solutions stored in the cylinder 3 and the cylinder 29a are spotted at a time by a predetermined amount on the surface of the substrate 53 via the respective dispensing needles 47.

In the initial stage of spotting, it is necessary to completely remove air from the cylinder 29a, the tube 35 and the dispensing needle 47 before spotting the sample solution. It is necessary to set the movement stroke. After that,
The operation plate 31 may be slightly moved by a stroke corresponding to the above-described predetermined amount to be dispensed.

The second upper and lower frames 41 of the dispensing device 5
After the substrate 53 on which the sample solution has been spotted is taken out after returning to the upper side, the second upper and lower frames 41 are lowered again so that the tips of the respective dispensing needles 47 are opposed to the substrate 53 of the next stage. The movement of the plate 31 is controlled in the same manner as described above,
A large number of sample solutions are spotted on the surface of No. 3 at a time by a predetermined amount. By repeating the above operation, the sample solution can be spotted on a large number of substrates 53 by a predetermined amount by one operation.

For example, a UV crosslinker (Hoef) is applied to the substrate 53 on which many sample solutions are spotted.
er UVC500) to dry the sample solution by irradiating it with ultraviolet light and fix it on the surface of the substrate 53.
The non-spotting region of the substrate 53 is immersed in a block solution (for example, succinic anhydride, N-methylpyrosinone, sodium borate solution, etc.) to be processed into a sample chip through a block treatment.

In the present embodiment, when a large number of sample solutions are spotted on the substrate 53, the sample solution is suctioned and a plurality of flow paths connected to a suction / discharge operating member for discharging a predetermined amount at a time are provided in each container body. A large number of suction flow paths for sucking the sample solution from the reservoir and a large number of discharge flow paths connected to each dispensing needle 47 can be selectively matched to suction and discharge a large number of sample solutions at once. In addition, the spotting operation of the sample solution can be performed efficiently.

Embodiment 2 FIG. 7 is an explanatory view showing a suction / ejection switching device according to this embodiment. In the first embodiment, the switching plate 17 is reciprocated by a predetermined stroke, and the suction port 19 and the discharge port 21 corresponding to each suction / discharge port 27 are selectively matched to enable communication. However, in this embodiment, as shown in FIG. 7, a large number of suction / discharge ports 27 are formed in the fixed plate 71 in a concentric or spiral shape. In addition, a large number of suction ports 19 are formed in the switching board 73 having the same size as the fixed board 71 so as to correspond to the respective suction / discharge ports 27, and each of the suction ports 1 is formed.
The discharge port portions 21 are formed at locations that are shifted by a predetermined angle in the circumferential direction from 9.

Then, the switching board 73 is moved by the electric motor 75 by the arrangement angle between the suction port 19 and the discharge port 21 described above.
By rotating, the suction port 19 and the discharge port 2 corresponding to each of the suction and discharge ports 27 in the fixed board 71.
1 and selectively communicate with each other, and the suction / ejection operating member 29
Thus, the sample solution can be suctioned and the sample solution can be discharged from the suction / discharge operation member 29.

In the above description, the fixed board 71 and the switching board 73 are constituted by one member. However, as shown in FIG. 8, at least a plurality of switching boards 81 (FIG. 8 shows three fixed boards and three switching boards). The configuration may be shown). The other configuration is the same as that of the first embodiment, and illustration and detailed description are omitted.

That is, the gear 8 connected to the electric motor 83
The three switching boards 81, which are divided into three parts, are rotatable in the same direction by meshing with each other, and a fixed board of the same size as each switching board 81 is provided on the upper surface side of each switching board 81. 8
7 are closely arranged. A number of suction and discharge ports 27 are formed at predetermined intervals on each fixed board 87, and a number of suction ports 19 and discharge ports 21 can be matched with each suction and discharge port 27 on each switching board 81. Formed.

Then, with the driving of the electric motor 83, each fixed plate 87 is rotated at an angle corresponding to the interval between the suction port 19 and the discharge port 21 in each of the switching boards 81 so as to be moved to each suction / discharge port 27. On the other hand, the suction port portion 19 and the discharge port portion 21 are selectively made to coincide with each other to enable the suction of the sample solution to the suction / discharge operation member 29 and the discharge of the sample solution from the suction / discharge operation member 29.

Embodiment 3 This embodiment relates to a suction / discharge switching apparatus using a valve instead of the switching board. That is, as shown in FIG. 9, each switching valve 91 having the same number as the spotting number has a three-port structure having a suction / discharge port port 93, a suction port 95, and a discharge port 97, and the suction / discharge port is set to common. .

A gear 99a is formed on an operating shaft 99 for switching a connection port in each switching valve 91, and each gear 99a is connected to a driving gear 1 connected to an electric motor 101.
03 are engaged so as to rotate in the same direction.

In accordance with the driving of the electric motor 101, the respective operating shafts 99 are rotated at a predetermined angle so that the suction port 95 and the discharge port 97 can be selectively communicated with the suction / discharge port 93 so that the suction / discharge is performed. The suction of the sample solution to the operation member 29 and the discharge of the sample solution from the suction / discharge operation member 29 are enabled.

The above description is based on the assumption that the fixing plate 1 is provided in the first holder 13.
5 and the switching plate 17 (the fixed plate 71 and the switching plate 73) are directly provided to switch the flow path between the suction / discharge operating member 29 and the suction needle 23 and the dispensing needle 47. A channel switching mechanism for switching the channels may be provided. In this case, the suction port 19 and the suction needle 23 and the discharge port and the dispensing needle 47 may be connected by respective tubes.

In the above description, a large number of substrates 53 are integrated in multiple stages on the substrate mounting table 51 and a large number of sample solutions are sequentially spotted on the uppermost substrate 53. A large number of substrates 53 may be arranged in a plane at predetermined intervals, and the dispensing needle group 49 may be controlled to move so that the sample solution is sequentially spotted on the many substrates 53 arranged in a plane. .

In the above description, the tip diameter of the dispensing needle is set to 100
However, in the present invention, the member for dispensing the sample solution onto the substrate may be a discharge nozzle, in which case it is a matter of course that the diameter is not limited to the above-mentioned tip diameter.

[0045]

According to the present invention, a large number of samples are spotted at high density on a substrate having a small occupied volume such as a slide glass, and at this time, a suction member and a dispensing member are connected to a suction / discharge operation member. It is possible to easily perform a spotting operation by simply switching a large number of flow paths to be used.

[Brief description of the drawings]

FIG. 1 is an explanatory view showing an outline of a spotting apparatus for implementing the present invention.

FIG. 2 is an explanatory diagram showing a suction / discharge device.

FIG. 3 is an explanatory diagram showing a switching mechanism.

FIG. 4 is an explanatory view showing a dispensing device.

FIG. 5 is an explanatory diagram showing a switching state.

FIG. 6 is an explanatory diagram showing a switching state.

FIG. 7 is an explanatory diagram illustrating a suction / ejection switching device according to a second embodiment.

FIG. 8 is an explanatory diagram showing a modified embodiment of the second embodiment.

FIG. 9 is an explanatory diagram illustrating a suction / ejection switching device according to a third embodiment.

[Explanation of symbols]

1-spotting device, 3-suction / discharge device, 5-dispensing device, 14-suction / discharge switching device, 15-fixed plate, 17-
Switching plate, 19-suction port forming suction flow path, 21-discharge port forming discharge flow path, 25-container body, 27-suction discharge port forming suction discharge flow path, 29-suction discharge Actuating member

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[Procedure amendment]

[Submission date] July 18, 2001 (2001.7.1)
8)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0006

[Correction method] Change

[Correction contents]

SUMMARY OF THE INVENTION The present invention has been made in order to solve the above-mentioned conventional drawbacks. The object of the present invention is to solve such a problem on a small occupied substrate such as a slide glass (26 mm × 76 mm). A sample that spots a large number of samples at high density, and at that time, easily switches a large number of flow paths connecting a suction member and a dispensing member to a suction / discharge operation member, thereby enabling efficient spotting operation. An object of the present invention is to provide a suction and discharge switching device for a solution spotting device.

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0012

[Correction method] Change

[Correction contents]

An air cylinder or a feed screw mechanism (not shown) is used as a drive mechanism for reciprocating the switching plate 17 at a distance corresponding to the distance between the suction port 19 and the discharge port 21.
Should be fine. In the above description, the movement of the switching plate 17 is controlled so that the suction port portions 19 are
And the discharge ports 21 are selectively matched to communicate with each other. However, the configuration may be such that the fixed plate 15 is moved and controlled to match.

[Procedure amendment 3]

[Document name to be amended] Statement

[Correction target item name] 0042

[Correction method] Change

[Correction contents]

The above description is based on the assumption that the fixing plate 1 is provided in the first holder 13.
5 and the switching plate 17 (the fixed plate 71 and the switching plate 73) are directly provided to switch the flow path between the suction / discharge operating member 29 and the suction needle 23 and the dispensing needle 47. A channel switching mechanism for switching the channels may be provided. In this case, the suction port 19 and the suction needle 23, and the discharge port 21 and the dispensing needle 47 may be connected by respective tubes.

[Procedure amendment 4]

[Document name to be amended] Drawing

[Correction target item name] Fig. 7

[Correction method] Change

[Correction contents]

FIG. 7

Claims (6)

[Claims] In a sample solution spotting apparatus for spotting a sample solution in a matrix on a substrate by a number of dispensing members, the sample solution stored in each of the reservoirs of the container is sucked and stored. For each of a number of suction / discharge channels communicating with each suction / discharge operation member that discharges a fixed amount of the sample solution to each dispensing member, each suction member for suctioning the sample solution stored in each reservoir of the container body. A switching member having a number of communication flow paths and a number of discharge flow paths communicating the suctioned sample solution to the respective dispensing members is moved to each suction flow path corresponding to each suction / discharge flow path. And a suction / discharge switching device in which the discharge passages are selectively matched to enable communication. 2. The switching member according to claim 1, wherein the connection port of each of the suction flow path and the discharge flow path is provided at a predetermined interval with respect to the fixed plate provided with the connection port of each suction and discharge flow path. Each of the formed switching plates is moved at a distance corresponding to the distance between the connection ports of the suction flow path and the discharge flow path, and the respective suction flow path and discharge flow path are selectively provided for each suction / discharge flow path. A suction / discharge switching device of a sample solution spotting device which is capable of communicating with the device. 3. The switching member according to claim 1, wherein the connection port of each of the suction flow path and the discharge flow path is provided at a predetermined interval with respect to the fixed plate provided with the connection port of each suction and discharge flow path. Each of the formed switching boards is rotated at an angle corresponding to the interval between the connection ports of the suction flow path and the discharge flow path to selectively connect the suction flow path and the respective suction flow path and the discharge flow path. A suction / discharge switching device of a sample solution spotting device which is capable of communicating with the device. 4. The suction / discharge switching device of a sample solution spotting device according to claim 3, wherein the switching plate comprises a single rotary plate connected to an electric motor. 5. The suction / discharge switching device of a sample solution spotting device according to claim 3, wherein the switching plate is rotatable in the same direction by meshing a plurality of rotating disks with a gear connected to an electric motor. 6. The switching member according to claim 1, wherein the number of switching members is equal to the number of channels provided with the connection port of the suction / discharge channel, the connection port of the suction channel, and the connection port of the discharge channel. A suction / discharge switching device for a sample solution spotting device, comprising a valve, wherein each switching valve selectively communicates each suction / discharge flow channel with each suction / discharge flow channel to enable communication.