JP2012020286A - Substance classifier and substance classification method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a substance classifier and substance classification method, which have a simple constitution, and enable a size classification with a high precision.SOLUTION: A passage 2 becoming gradually shallower toward the downstream side while inclining the bottom is formed on a glass plate (passage substrate) 1. A cover glass (plate) 3 covers so as to occlude a part of the passage 2 in the passage direction. At least the cover glass 3 among the glass plate 1 and the glass plate 1 is made of a transparent member. The flow passage 2 is continuously aslant toward the downstream side. Scale marks m-mare attached to the cover glass 3 at a predetermined interval along the passage.

Description

  The present invention is used for collecting or detecting fine particles and impurities in a solution, and particularly suitable for classifying biological materials such as cells, bacteria, biopolymers (for example, proteins and DNA) and the like. It relates to a substance classification method.

  As a technique for classifying substances in a solution, for example, techniques shown in Patent Documents 1 to 3 are known. In Patent Document 1, particles are selected by applying an alternating electric field to particles of different sizes accommodated in a microchannel. Patent Document 2 describes a method in which electrodes are arranged in microchannels and particles are selected based on the charge characteristics of the particles. Patent Document 3 describes a method of classifying particles by forming different laminar flows according to the size of particles in a flow path of a solution containing particles of different sizes.

JP 2004-243238 A JP-A-5-126696 JP 2005-205387 A

However, in the methods of Patent Documents 1 and 2 that use an electric field, it is necessary to provide an electrode for forming an electric field in the flow path, and classification is performed depending on the charge characteristics of the particles. When intended, accurate results could not be obtained. There is also a problem that substances that can be classified are limited to charged particles.
In addition, the method disclosed in Patent Document 3 complicates the flow path structure, requires two systems of fluid supply sources (pumps) to form a flow, and has a flow rate and flow rate. Strict control is required.

  The present invention has been made in view of the above-described problems, and an object of the present invention is to provide a substance classification apparatus and a substance classification method capable of size classification with a simple configuration and high accuracy.

  In order to solve the above-mentioned problems, a substance classification device according to claim 1 of the present invention includes a flow path substrate in which a concave flow path extending along a surface is formed, and is superimposed on the flow path substrate. A plate that covers a part of the flow path direction, and the flow path becomes shallower toward the downstream side, and traps the substance flowing in the flow path between the plate and the bottom surface of the flow path. A material classifying device for forming a collecting space, wherein at least the plate is a transparent member of the flow channel substrate and the plate, and the flow channel is continuously inclined toward the downstream side. A scale that is attached along the direction and can specify the size of the substance corresponding to the position in the flow path direction where the substance is collected is provided.

  In the substance classifier having such a configuration, the diameter of the substance in the solution is approximately equal to the distance between the plate and the bottom surface of the channel (hereinafter referred to as “gap”). Be collected. Since the gap has a size corresponding to the position in the flow path direction, the size of the substance to be collected can be specified by the position in the flow path direction where the substance is collected. It is the size of the gap that directly correlates with the size of the substance, but the scale along the channel direction is easier to see when the channel substrate is placed horizontally, and in many cases the gap Since the length of the collection space in the flow path direction is larger than the size, the accuracy of determining the size is increased.

The substance classification apparatus according to a second aspect of the present invention is the substance classification apparatus according to the first aspect, wherein the scale displays a collection position according to the size of the substance.
A substance classifying apparatus according to claim 3 of the present invention is the substance classifying apparatus according to claim 1 or 2, wherein the transparent material is glass.
The substance classification apparatus according to claim 4 of the present invention is the substance classification apparatus according to any one of claims 1 to 3, wherein the flow path has a bottom surface inclined with respect to the substrate surface of the flow path substrate. It is formed as a surface.

A substance classification device according to a fifth aspect of the present invention is the substance classification device according to any one of the first to third aspects, wherein the scale is a flow based on a downstream end of the flow path. It is attached to an arbitrary position in the road direction.
A substance classification device according to a sixth aspect of the present invention is the substance classification device according to any one of the first to third aspects, wherein the scale is attached to the plate or the flow path substrate. And

A substance classification device according to a seventh aspect of the present invention is the substance classification device according to any one of the first to sixth aspects, further comprising a slide mechanism that enables the plate to slide along the flow path direction. It is characterized by that.
The substance classification apparatus according to an eighth aspect of the present invention is the substance classification apparatus according to the seventh aspect, wherein the slide mechanism includes a drive device that drives the plate along the flow path direction. To do.

The substance classifying apparatus according to claim 9 of the present invention is the substance classifying apparatus according to claim 7, wherein the slide mechanism includes a guide portion that guides the plate along the flow path direction. To do.
In the substance classification method according to claim 10 of the present invention, a flow path substrate in which a concave flow path extending along the surface is formed, and a part of the flow path in the flow path direction are overlapped with the flow path substrate. A material classifying device that forms a collection space for a substance that flows in the flow path between the plate and the bottom surface of the flow path, the flow path being shallower toward the downstream side. a substance classification method using, together with at least said plate is a transparent member of the front Kiryuro substrate and the plate, and continuously tilting the flow passage toward the downstream side, said material The size of the collected substance is specified based on the collected position in the channel direction.
A substance classification method according to an eleventh aspect of the present invention is the substance classification method according to the tenth aspect, wherein the plate is moved to a position where the collected target substance is released after the substance is collected in the collection space. The target substance is recovered by sliding the plate to the upstream side.

  According to the present invention, highly accurate size classification can be performed with a simple apparatus.

(A) is a side view of the substance collection apparatus of this embodiment, (b) is a top view. It is a figure which shows the method of forming a flow path in a substance collection apparatus. FIG. 3 is a diagram for explaining the structure of a flow path 2. (A) is a side view of the substance collection apparatus of 2nd Example, (b) is a top view at the time of sliding the cover glass of the substance collection apparatus of (a) to an upstream side. (A) is a side view of the material collection device of the third embodiment, (b) is a plan view when the cover glass 3 of the material collection device of (a) is slid upstream, and (c) is ( It is a top view of the state which removed the cover glass of the substance collection apparatus of a). It is a figure for demonstrating the use condition of the base with a scale line.

Next, Examples 1 to 3 which are embodiments of the present invention will be described with reference to the drawings.
[First embodiment]
(Constitution)
FIG. 1A is a side view of the substance collection device A of the first embodiment, FIG. 1B is a plan view, and FIG. 2 is a diagram showing a method for forming the flow path 2 in the substance collection device A. 3 is a diagram for explaining the structure of the flow path 2.
The substance collection apparatus A shown in FIG. 1 is formed by overlapping a cover glass 3 on a glass plate (10 × 20 mm, thickness 1 mm) 1 on which a flow path 2 is formed. The glass plate 1 corresponds to the flow path substrate of the present invention, and the cover glass 3 corresponds to the plate of the present invention.
The flow path 2 was produced as shown in FIG. First, a Si wafer having a thickness of 500 μm was laid on the back side of one end portion in the longitudinal direction of the glass plate 1, and the glass plate 1 was fixed while being inclined with respect to the horizontal plane. At this time, the glass plate 1 is fixed at an inclination of about 1.43 ° (= Tan (0.5 / 20)). Thereafter, using a dicing saw, a straight groove having a width of 100 μm was formed in the longitudinal direction of the glass with a cutting amount of 300 μm from the side on which the Si wafer was laid. As a result, a straight groove-like channel 2 having a bottom surface inclined by about 1.43 ° along the channel direction is formed on the surface of the glass plate 1 with a length of about 12 mm.

A commercially available cover glass (8 × 12 mm, thickness 0.12 to 0.17 mm) is used for the cover glass 3, and the channel end 2 a of the channel 2 (the end of the channel 2 whose bottom is shallower) And fixed upstream at a position of 0.2 mm. When the inclination of the bottom surface of the flow path 2 is about 1.43 °, the depth of the flow path 2 and the distance from the flow path end 2a are in the relationship shown in Table 1. For this reason, the gap G between the bottom face of the flow path 2 and the downstream end of the cover glass 3 is about 5 μm (= 0.2 × (Tan1.43 °)) (see FIG. 3). In addition, scale lines m _{1 to} m ₅ were attached to the cover glass 3 at predetermined intervals (for example, 1.5 mm intervals) along the flow path direction.

  A tube 5 (outside diameter of about 300 μm) for introducing the sample solution was fixed to the flow path start end 2b side, and the periphery of the tube 5 was sealed with PDMS (polydimethylsiloxane) so that the sample solution did not leak. Further, on the side of the flow path end 2a on the glass plate 1, the flow path end 2a is surrounded by a side wall 4a made of PDMS, thereby forming a recovery chamber 4 for recovering waste liquid.

(Function and effect)
Next, the usage method and effect of the substance collection apparatus A having the above configuration will be described.
When the substance collection device A is used, the sample solution is sent out from the flow path start end 2b to the flow path 2 by a pump. In the middle of the flow path 2, among the substances in the sample solution, a substance having substantially the same diameter as the gap G between the cover glass 3 and the bottom surface of the flow path 2 is sandwiched and collected. The size of the gap G corresponds to the depth of the flow path 2 and becomes smaller toward the downstream side due to the inclination of the bottom surface of the flow path 2, so that a substance having a smaller size is collected on the downstream side, As shown in Table 1, the position in the flow path direction correlates with the size of the collected substance. Accordingly, as shown in FIGS. 1A and 1B, when substances of various sizes are flowed, the trapping substance s is distributed in the flow direction according to the size, and the size of the substance is It can be known from the scale lines of the cover glass 3 attached at predetermined intervals along the flow path direction. For example, in the case of the relationship shown in Table 1, it can be confirmed that the substance in the vicinity of the graduation line m _{2 provided} at a position of 3.2 mm from the channel end 2a has an outer diameter of about 80 μm.

  After flowing the sample solution, the glass plate 1 and the cover glass 3 are transparent members, so that the target size substance can be observed with a microscope while the glass plate 1 is covered with the cover glass 3. You may take out the substance s of the target size from the collection material s. When taking out the target substance s, the liquid feeding from a pump is stopped and the cover glass 3 is removed. At this time, if the solution remaining between the glass plate 1 and the cover glass 3 is sucked and removed from the downstream side with a dropper or the like and then removed, when the cover glass 3 is removed, the downstream collection substance s flows out to the upstream side at the time of removal. The target substance can be taken out while preserving the state at the time of collection. Moreover, when the residual liquid is removed by turning the glass plate 1 upside down, the state at the time of collection is maintained as shown in FIG. 1 (c), and the collection is performed on the cover glass 3 side with the graduation line m attached. Since the substance s can be taken out, it can be taken out in a state where the size is specified accurately.

Note that a substance having an outer diameter smaller than the gap G flows to the flow path end 2 a and is collected in the collection chamber 4.
As described above, since the size of the collected substance can be specified by attaching a scale line corresponding to the position in the flow path direction along the flow path direction of the cover glass 3, the mixed solution of various substances is classified. Or the size of the target substance can be specified, or the target size substance can be separated. It is the size of the gap that directly correlates with the size of the material, but it is easier to see the scale along the flow path direction because the flow path substrate is placed horizontally, as compared to the size of the gap. In addition, since the length of the collection space in the flow path direction is larger, the accuracy of determining the size becomes higher.

In the first embodiment, the scale lines m _{1 to} m ₅ are attached at predetermined intervals along the flow path direction. However, the present invention is not limited to this, for example, only in the flow path direction position according to the size of the target substance. A scale line may be attached. In the first embodiment, only the scale line is attached, but a numerical value of the outer diameter size of the substance specified by the position of the scale line may be attached together with the scale line.

[Second Embodiment]
Next, a second embodiment will be described.
FIG. 4 shows a substance collection apparatus A of the second embodiment (FIG. 4A is a side view, and FIG. 4B is a slide of the cover glass 3 of the substance collection apparatus A of FIG. 4A to the upstream side. Is a plan view of the case). The substance collection device A of the second embodiment has substantially the same configuration as that of the first embodiment, but the piezoelectric element which is the driving device of the present invention is connected to the cover glass 3 and the cover glass 3 is directed in the flow path direction. It is different in that it can slide along. Hereinafter, the description will be focused on the different points, and description of the other parts will be omitted.

  In the substance collection device A of FIG. 4, ribs 6 protruding in the thickness direction of the glass plate 1 are formed along the periphery on the surface side of the glass plate 1. The rib 6 is not formed on one end side in the short direction of the glass plate 1 and on the upstream side of the flow path 2. Further, the portion formed on the opposite end side forms a recovery chamber 4 for the solution flowing out from the channel end 2a. The cover glass 3 is superimposed on the surface of the glass plate 1 in a state of being connected to the piezoelectric element as described above, guided by the rib 6, and slidable along the flow path direction. The rib 6 corresponds to the guide portion of the present invention, and can be formed of, for example, a resin material, a metal material, or glass. The slide mechanism of the present invention is constituted by the rib 6 and the piezoelectric element. The structure and forming method of the flow path 2, the material and size of the glass plate 1 and the cover glass 3, and the fact that scale lines with a predetermined interval are attached to the cover glass 3 are the same as in the first embodiment. .

Next, the usage method and effect of the substance collection apparatus A having the above configuration will be described.
When using the substance collection device A, the cover glass 3 is driven, and the position of the cover glass 3 is set at a position where the substance can be collected according to the size of the target substance. In other words, in the second embodiment, by changing the position of the cover glass 3, the collection of substances of various sizes can be handled by the single substance collection device A. After setting the position of the cover glass 3, the sample solution is pumped out. As a result, as in the first embodiment, the substance s having the same diameter as the gap is collected between the cover glass 3 and the bottom surface of the flow path 2 (see FIG. 4A). The size of the trapping substance s can be confirmed by the scale lines m _{1 to} m ₅ attached to the cover glass 3. In the second embodiment, the position from the channel end 2a is changed by adjusting the position of the cover glass 3. Therefore, after the cover glass 3 is adjusted, it is necessary to confirm the position from the channel end 2a. is there.

After flowing the sample solution, the glass plate 1 and the cover glass 3 are transparent members, so that the target size substance can be observed with a microscope while the glass plate 1 is covered with the cover glass 3. You may take out the substance s of the target size from the collection material s.
In order to take out the target substance s, liquid feeding from the pump is stopped, and the cover glass 3 is slid to a position where the target collection substance s is released by driving with a piezoelectric element. This will be specifically described with reference to FIG. FIG. 4B shows an example in which a target substance whose size is specified in advance is collected from a sample solution. In this case, before flowing the sample solution, the position of the cover glass 3 is set so that the target substance s1 (black circle in the figure) is collected near the downstream end of the cover glass 3. After flowing the sample solution, the cover glass 3 is slid to the position where the collected target substance s1 is released, and then fed again from the pump as it is, so that only the released target substance s1 is in the collection chamber. 4 is sent out. Thereby, the target substance s1 can be recovered from the recovery chamber 4. Instead of pumping liquid from the upstream side, the target substance s1 may be recovered by sucking the solution from the downstream side with a dropper or a pump.

As described above, by making the cover glass 3 slidable along the flow path direction, it is possible to collect substances of various sizes, and easily collect substances of the desired size from the collected substances. it can.
In the second embodiment, the case where only the trapping substance s1 is recovered has been described. However, if the above operation is repeated, the trapping substances s2 to s4 can be sequentially recovered from the small size to the large one.
Further, the method of sliding the cover glass is not limited to the piezoelectric element, and a method (actuator) corresponding to the size (slide distance) of the target substance can be used.

[Third embodiment]
Next, a third embodiment will be described.
FIG. 5 shows a substance collection apparatus A of the third embodiment (FIG. 5A is a side view, and FIG. 5B is a slide of the cover glass 3 of the substance collection apparatus A of FIG. 5A to the upstream side. (C) is a plan view with the cover glass 3 removed). The material collection device A of the third embodiment is different from the second embodiment in that the scale lines m _{1 to} m ₅ are attached not to the cover glass 3 but to the glass plate 1. The scale lines m _{1 to} m ₅ are attached to the rear surface side of the glass plate 1 by a method such as printing or etching at a predetermined distance from the channel end 2a. The scale lines m _{1 to} m ₅ can be confirmed even from the front side because the glass plate 1 and the cover glass 3 are transparent. Since other parts are the same as those of the second embodiment, description thereof is omitted.

As described above, when the scale lines m _{1 to} m ₅ are attached to the glass plate 1 side, the scale lines m _{1 to} m ₅ do not move even if the cover glass 3 is slid after the sample solution has finished flowing. The collected material s1 can be recovered by accurately grasping the size (see FIG. 5B).
If the solution remaining in the flow path 2 is sucked and removed from the downstream side with a dropper or the like, the size-classified state can be maintained even if the cover glass 3 is completely removed as shown in FIG. The target substance may be recovered from the state.
Further, as shown in FIG. 6, the size may be determined by laying a pedestal 7 with a graduation line under the glass plate 1.

DESCRIPTION OF SYMBOLS 1 Glass plate 2 Flow path 2a Flow path end 2b Flow path start end 3 Cover glass 4 Collection chamber 4a Side wall 5 Tube 6 Rib A Material collection device G Gap m Scale line

Claims (11)

A flow path substrate formed with a concave flow path extending along the surface;
A plate overlaid on the flow path substrate and covering a part of the flow path direction of the flow path,
The flow path is shallower toward the downstream side, and is a substance classification device that forms a collection space for a substance flowing in the flow path between the plate and the bottom surface of the flow path,
While at least the plate of the flow path substrate and the plate is a transparent member,
The flow path is continuously inclined toward the downstream side,
A substance classification apparatus comprising a scale attached along a flow path direction and capable of specifying a size of the substance corresponding to a position in the flow path direction where the substance is collected.   2. The substance classification apparatus according to claim 1, wherein the scale displays a collection position according to the size of the substance.   3. The substance classification apparatus according to claim 1, wherein the transparent material is glass.   The substance classification device according to any one of claims 1 to 3, wherein a bottom surface of the flow path is formed as an inclined surface with respect to a substrate surface of the flow path substrate.   4. The substance classification device according to claim 1, wherein the scale is attached to an arbitrary position in a flow path direction with respect to a downstream end portion of the flow path. 5. .   The substance classification device according to any one of claims 1 to 3, wherein the scale is attached to the plate or the flow path substrate.   The substance classification apparatus according to any one of claims 1 to 6, further comprising a slide mechanism that allows the plate to slide along a flow path direction.   The substance classification device according to claim 7, wherein the slide mechanism includes a drive device that drives the plate along the flow path direction.   The substance classification device according to claim 7, wherein the slide mechanism includes a guide unit that guides the plate along the flow path direction. A flow path substrate formed with a concave flow path extending along the surface;
A plate overlaid on the flow path substrate and covering a part of the flow path direction of the flow path,
The flow path is shallower toward the downstream side, and is a material classification method using a material classification device that forms a collection space for a substance flowing in the flow path between the plate and the bottom surface of the flow path. There,
Together with at least said plate is a transparent member of the front Kiryuro substrate and the plate,
The flow path is continuously inclined toward the downstream side,
A substance classification method, wherein the size of the collected substance is specified based on the position in the flow path direction where the substance is collected.   The material according to claim 10, wherein after collecting the substance in the collection space, the target substance is collected by sliding the plate upstream to a position where the collected target substance is released. Substance classification method.

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