JP2009145097A - Chemical material identifying sensor and chemical material identifying method using it - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a chemical material identifying sensor for surely arranging, at a predetermined portion, a bead which has caused hybridization with a target material such as many chemical materials, with no need of a chemical bonding portion, and to provide a measuring method using it. <P>SOLUTION: This sensor includes a plate 1 where a recess 2 having a through-hole 3 is arranged and a first and a second region 4, 5 which are divided by the plate 1. While a micro particle 11 having a probe function is disposed at the recess 2 of the through-hole, a liquid is filled in one of the first, the second region 4, 5 and in the through-hole 3. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a chemical substance identification sensor capable of measuring a sample such as a chemical substance with high accuracy and an identification method using the same.

  Conventionally, as a method for measuring a protein or the like present in a solution, a chemical substance identification sensor having a protein probe on the surface of a bead is known. For example, this chemical substance identification sensor is a probe that is a bioactive agent on the surface of a bead. There is known a technique for detecting a target substance that adsorbs and hybridizes with a probe portion in a solution. As an example of this chemical substance identification sensor, a technique using a substrate / bead pairing that can bind or attach a bead to a separated site on the surface of the substrate so that the bead does not move during measurement is disclosed. (For example, refer to Patent Document 1).

Special Table 2002-533727

  However, in the configuration of the conventional chemical substance identification sensor, the beads are arranged at a specific portion of the substrate, so that the beads are arranged by a method such as mixing, vibration, or stirring, and the chemical substance is bonded to fix the substrate. However, the method described above does not always allow beads to be arranged at all sites of the array, and conversely has a problem that a plurality of beads are arranged in one array.

  The present invention solves the above-mentioned conventional problems, and it is possible to reliably place beads hybridized with a target substance such as a large number of chemical substances at a predetermined site, and the need for a chemical binding site. It is an object of the present invention to provide a chemical substance identification sensor having no defect and a measurement method using the same.

  In order to solve the above problems, the present invention includes a plate provided with a through hole having a depression and first and second regions partitioned by the plate, and the depression of the through hole has a probe function. The microspheres are arranged and the inside of any one of the first and second regions and the inside of the through hole are filled with a liquid.

  The above configuration realizes a sensor structure in which the beads can be easily arranged in a recess provided in a part of the through hole, and thus it is necessary to chemically bond or attach the beads to a specific portion of the substrate. Thus, it is possible to provide a chemical substance identification sensor capable of reliably fixing one bead to a recess provided in the upper part of the through hole, and a measurement method using the same.

(Embodiment 1)
Hereinafter, the configuration of the chemical substance identification sensor according to the first embodiment of the present invention will be described with reference to the drawings. FIG. 1 is a partially cutaway perspective view of a chemical substance identification sensor of the present invention, and FIG. 2 is a cross-sectional view thereof. 1 and 2, reference numeral 1 denotes a plate made of silicon, and a part of the plate 1 is a thin portion 1a.

  Furthermore, the first region 4 and the second region 5 are communicated only by a through hole 3 having a recess 2 provided in the thin portion 1a. In addition, a light entrance 8 and a light detection port 9 such as an optical fiber are respectively installed in the first region 4 and the second region 5, and the light entrance 8 and the light detection port 9 are further measured. Connected to a vessel (not shown).

  Further, the inside of the through hole 3 having the first region 4, the second region 5, and the recess 2 is filled with the liquid 10. Here, the liquid 10 is a liquid substance in which the target chemical substance to be measured exists. Further, caps 6 and 7 are brought into contact with the first region 4 and the second region 5 as an example of a cover plate, thereby isolating the first region 4 and the second region 5 from the external atmosphere. As a result, the internal pressure can be controlled.

  In the present embodiment, the first region 4 and the second region 5 are both filled with the liquid 10, but the light that has passed through the through hole 3 to the second region 5 is thereby in the second region 5. Since the liquid 10 satisfying the above condition is diffused, there is an advantage that introduction into the light detection port 9 installed on the second region 5 side becomes easy. However, on the other hand, if it is not preferable to diffuse light in the second region 5, the second region 5 side may not be filled with liquid. In this case, the light that has passed through the through-hole 3 reaches the light detection port 9 directly.

  Then, particles (beads) 11 are arranged in the recesses of the depressions 2, and chemical substances are identified through the particles 11. The material of the particles (beads) 11 is not particularly limited, but preferably has transparency with respect to the excitation light 14 and the detection light 15, and has such transparency. Suitable materials include inorganic materials such as glass and ceramic materials, or resin materials such as polycarbonate, polystyrene, and polyolefin.

  Further, the size of the particles 11 is a spherical shape having a diameter of about 0.5 to 100 μm, so that the particles 11 can be stably held inside the recess 2 provided in the plate 1 of the present invention. However, the shape other than the spherical shape is not excluded. For example, in order to increase the adhesion amount of the probe 12 to the surface, the present invention can be applied even to an irregular shape having irregularities on the surface.

  A probe 12 is attached to the surface of the particle 11, and this probe 12 is synthesized as a specific compound corresponding to the detection target substance 13. For example, a DNA probe is a DNA having a complementary base sequence that binds to a specific base sequence. When it is desired to determine the presence or absence of DNA having a specific sequence in a solution, it binds to this DNA probe (hybridization). Can be measured by whether or not

  In particular, in the present invention, the probe 12 is fluorescently labeled, and when the detection target substance 13 is bound, fluorescence is emitted to the excitation light 14 that is input light from the outside. As a result, the color of the detection light 15 that has passed through the surface of the particle 11 changes depending on the presence or absence of hybridization, but with the sensor of the present invention, only the detection light 15 that has passed through the through-hole 3 can be observed. Therefore, it is possible to reduce noise from others, and this makes it possible to identify chemical substances with higher accuracy.

  In addition, RNA, protein, etc. other than DNA can be used for the type of probe 12.

  Next, a method for identifying a chemical substance present in the liquid 10 using the chemical substance identification sensor of the present invention will be described. 3 and 4 are cross-sectional views showing a method of using the chemical substance identification sensor of the present invention. Here, in the first region 4 of the chemical substance identification sensor according to the first embodiment shown in FIG. 3, the particles 11 having the probe 12 formed on the surface are held inside the recess 2 of the plate 1. . Here, since the opening of the through-hole 3 is smaller than the particle 11, as a method for holding the particle 11, the second region 5 side is decompressed and the through-hole 3 is sucked to easily realize strong holding. Is done. In addition, since this holding method uses the pressure difference between the first region 4 and the second region 5, there is no need to separately provide a chemical bonding site for adsorbing the particles 11 in the recess 2. have.

  Moreover, since the 1st area | region 4 and the 2nd area | region 5 are partitioned off with the plate 1, by pressing the inside of the 1st area | region 4 with a pressurization means (not shown), the through-hole 3 is made. Accordingly, the particles 11 can be held in the depressions 2 while moving the particles 11 into the depressions 2 while moving the liquid 10 to the second region 5.

  In this way, by adopting a configuration in which the first region 4 and the second region 5 can be individually controlled, the particles 11 can be easily controlled using means such as suction or pressurization. Furthermore, it is possible to realize a sensor structure that allows the liquid 10 to be easily replaced with another chemical solution or the like.

  Next, when the excitation light 14 is introduced from the first region 4 side as shown in FIG. 3, the light is transmitted while being scattered inside the liquid 10 filling the inside of the first region 4, as shown in the figure. It also enters the inside of the through hole 3 having the depression 2. At this time, since the particles 11 held in the depressions 2 reliably block the through holes 3, the excitation light 15 transmitted through the through holes 3 is light transmitted through the surfaces of the particles 11. As shown in the figure, when the inspection target substance 13 is not present in the solution 4 or the concentration is low, the probe 12 formed on the surface of the particle 11 does not hybridize with the inspection target substance 13, and this state The light passing through the surface of the particle 11 is detected as the detection light 15 and the reference response is measured.

  Next, as shown in FIG. 4, in the state where the inspection target substance 13 is present in the solution 4, the probe 12 is hybridized with the inspection target substance 13, and the detection light that has passed through the surface of the particle 11 in this state 15 is detected. At this time, in the state where the probe 12 and the substance to be inspected 13 are hybridized, if the probe 12 is modified so as to emit fluorescence, the detected light 15 that has passed passes through the fluorescence, and the reference response Distinguished from In this way, it is possible to determine whether or not the inspection target substance 13 exists in the liquid 4.

  Thus, by providing the depression 2 having the through hole 3 in the plate 1, it is possible to reliably hold the particle 11 having the probe 12 on the surface, and light that has passed through the through hole 3 is always on the surface of the particle 11. Since it passes through, more accurate measurement is possible. Therefore, the material of the plate 1 can be other than the silicon described in the first embodiment, but is more preferably opaque. As a result, only the light transmitted to the second region 5 side passes through the through hole 3, and the light detection on the second region 5 side becomes more accurate. The second region 5 on the detection side can be detected even if it is not necessarily filled with liquid, but when it is filled with liquid, the light that has passed through the through hole 3 exists in the second region. It has the advantage that it is scattered inside the liquid 10 and can be easily introduced into the light detection port 9.

  Furthermore, in the holding method according to the first embodiment, if the first region 4 side is in a reduced pressure atmosphere, the holding of the particles 11 once held can be released again. It is possible to promote the reaction between the probe 12 and the substance 13 to be inspected. For example, after the reference response is measured by holding the particle 11 in the depression 2 without the inspection target substance 13 in the liquid, the liquid 10 containing the inspection target substance 13 is added, and then the holding of the particle 11 is released. For example, the particles 11 are easily dispersed in the liquid 10 to promote the reaction between the substance to be inspected 13 and the probe 12, and if the measurement is performed after the particles 11 are held again by suction, the object to be inspected is compared with the reference response. There is an advantage that the presence / absence determination of the substance 13 becomes more accurate.

  Further, the plate 1 and the thin portion 1a are made of silicon, whereby the processing of the recess 2 and the through hole 3 formed inside the plate 1 and the thin portion 1a is efficiently performed by using a semiconductor process. And it becomes possible to produce with high precision.

  In addition, a more reliable measurement is possible by providing a plurality of through holes 3 having the recesses 2 provided in the thin portion 1a.

  As described above, the chemical substance identification sensor described in the first embodiment and the chemical substance identification method using the same are a DNA sensor that detects a specific DNA sequence such as a virus or a food production area, and an SNP. (Single nucleotide polymorphisms) SNP sensors for detecting sequences, antigen sensors for detecting the presence of allergens (allergic antigens), and the like can be widely used in the agricultural field, medical field, environmental field, and the like.

(Embodiment 2)
A chemical substance identification sensor according to Embodiment 2 of the present invention will be described with reference to the drawings.
As shown in FIG. 5, it is possible to adopt a structure in which one side of the plate 1 is opened to the outside atmosphere. In this case, by attaching a chemical substance identification sensor directly to the liquid 10, The presence or absence can be measured.

  In the second embodiment, the first region 4 is an open space, and the second region 5 is closed from the outside atmosphere except for the pressure control port 16. In this case, since the first region 4 side is an open space, in order to control the pressure on the first region 4 side, the sinking amount from the liquid level of the chemical substance identification sensor to be submerged into the liquid 10 is set. Control is sufficient. That is, if this is submerged deeply, the particles 11 can be held in the through holes 3 with a stronger pressure. When combined with the pressure control of the second region 5 by the pressure control port 16 connected to the second region 5, the particles 11 once held in the depressions 2 are made by making the second region 5 side a positive pressure. It is also possible to disperse again.

  Further, since the first region 4 is an open space, there is an advantage that the exchange of the particles 11 dispersed on the first region 4 side, the exchange of the liquid 10 and the like are easier.

  As described above, the chemical substance identification sensor and the chemical substance identification method using the same according to the present invention are useful for, for example, the use of detecting the presence or absence and concentration of the test target substance present in the liquid with high accuracy. .

1 is a partially cutaway perspective view of a chemical substance identification sensor according to Embodiment 1 of the present invention. Cross section of the chemical substance identification sensor Sectional view showing how to use the chemical substance identification sensor Sectional view Sectional drawing of the chemical substance identification sensor in Embodiment 2 of this invention

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Plate 1a Thin part 2 Indentation 3 Through-hole 4 1st area | region 5 2nd area | region 6 Cap 7 Cap 8 Light inlet 9 Light detection port 10 Liquid 13 Test object substance 14 Excitation light 15 Detection light 16 Pressure control port

Claims (10)

  A plate provided with a through hole having a depression, and first and second regions partitioned by the plate, and a microsphere having a probe function is disposed in the depression of the through hole, and the first, A chemical substance identification sensor in which a liquid is filled in any one of the second regions and the inside of the through hole.   The chemical substance identification sensor according to claim 1, wherein a liquid is filled in the first area and the second area.   The chemical substance identification sensor according to claim 1, wherein the pressures in the first region and the second region are individually controlled.   The chemical substance identification sensor according to claim 1, wherein the plate is opaque.   The chemical substance identification sensor according to claim 1, wherein the plate is made of silicon.   The chemical substance identification sensor according to claim 1, wherein the opening diameter of the through hole is smaller than the outer shape of the bead.   The chemical substance identification sensor according to claim 1, further comprising suction means for sucking beads by sucking from a through-hole having no depression.   The chemical substance identification sensor according to claim 1, further comprising a pressurizing means for placing the beads in the depression by pressurizing the first region on one side of the plate in which the through hole having the depression is formed.   The chemical substance identification sensor according to claim 1, wherein the chemical substance identification sensor is formed on the surface of a microsphere by being modified to emit fluorescence by binding to a chemical substance to be detected as a probe function.   A plate provided with a through hole having a depression, and first and second regions partitioned by the plate, and a microsphere having a probe function is disposed in the depression of the through hole, and the first, Using a chemical substance identification sensor in which liquid is filled in the inside of the second region and the inside of the through hole having the depression, light is irradiated from one of the two regions, and the depression is formed from the other region. A method for identifying a chemical substance that measures the wavelength and amount of light that has passed through a through-hole.

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