WO2021199926A1 - Outil de collecte de micro-échantillon et dispositif de collecte - Google Patents

Outil de collecte de micro-échantillon et dispositif de collecte Download PDF

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Publication number
WO2021199926A1
WO2021199926A1 PCT/JP2021/008896 JP2021008896W WO2021199926A1 WO 2021199926 A1 WO2021199926 A1 WO 2021199926A1 JP 2021008896 W JP2021008896 W JP 2021008896W WO 2021199926 A1 WO2021199926 A1 WO 2021199926A1
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WIPO (PCT)
Prior art keywords
sample piece
collecting tool
protrusion
analyzed
micro
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PCT/JP2021/008896
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English (en)
Japanese (ja)
Inventor
健嗣 和田
亨 篠原
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野村メディカルデバイス株式会社
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Publication of WO2021199926A1 publication Critical patent/WO2021199926A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting

Definitions

  • the present invention relates to a micro sample piece collecting tool and a collecting device.
  • the method of collecting a minute sample piece used for elemental analysis of an object or genome analysis of a living tissue is that a person grasps an object by grasping a metal rod with a sharp tip such as tweezers or a probe, or a collecting tool such as a sharp spoon. It was common to collect minute sample pieces by scraping them from living tissue. However, such a collection method has a problem that contaminants are easily mixed in the collected microsections and are not suitable for analysis.
  • a collection device in which a micro sample piece in which specific cells are present is collected from a thin section of a biological tissue to be analyzed and used for various analyzes.
  • a plurality of microchips arranged at a predetermined distance (gap portion) from each other are prepared, and the entire surface of the upper surface (mounting surface) of the plurality of microchips is covered.
  • a thin section of the biological tissue is arranged, and the thin section is melt-attached to the mounting surface by raising the temperature, and the thin section is momentarily shrunk by further heating to make it thin in the gap between the microchips.
  • a sampling device has been proposed in which a section is cut and pushed up one by one with a movable pin to separate a minute sample piece from the thin section and collect it.
  • Patent Document 2 a thin section of biological tissue is attached to the surface of a substrate that can be divided into a plurality of small pieces, and the substrate to which the thin section is attached is adhered to a sheet-shaped extension member to form an extension member.
  • a sampling device has been proposed in which a minute sample piece is separated from a thin section and collected by extending it in all directions to widen the gap between small pieces of the substrate.
  • the sampling device described in Patent Document 1 has a configuration in which thin sections are attached on a plurality of minute chips, the minute chips are pushed up by corresponding movable pins for each minute chip, and the minute sample pieces are separated.
  • the sampling device described in Patent Document 2 has a configuration in which a thin section is attached to an extension member and then the extension member is extended to separate minute sample pieces. Therefore, in both the sampling devices of Patent Document 1 and Patent Document 2, in the process of manufacturing the component parts constituting the mechanism and assembling the sampling device, dust, dust due to wear of the members, processing oil, and an operator There is a high possibility that the oils and fats, body fluids, etc. of the above have adhered to and remain on the sampling device in some way.
  • sampling devices in the sampling process, a plurality of steps are required to collect the micro sample pieces, so that there is a considerable chance that contaminants are mixed in the micro sample pieces in the sampling process. , There is a risk that it will not be possible to provide minute sample pieces suitable for analysis.
  • the present invention has been made to solve at least one of such problems, and it is possible to collect minute sample pieces from various objects to be analyzed, not limited to living tissues, and the minute amount is small.
  • the micro sample piece collecting tool of the present invention is a micro sample piece collecting tool for collecting a part of an object to be analyzed as a micro sample piece, and is integrated with a base made of an inorganic solid material and the base. With the tip of the viscoelastic body pierced or pressed against the object to be analyzed, the viscoelastic body moves relative to the object to be analyzed. It is characterized by having a protrusion for scraping a part of the object to be analyzed.
  • the inorganic solid material is a silicon substrate and the protrusions are formed by using a photolithography technique.
  • the protrusion is composed of a cone, a thruster, or a pillar.
  • the protrusion when the protrusion is composed of a pillar, the protrusion has a tapered shape in which the area of the bottom surface of the pillar is smaller than the area of the top surface. It is preferable to have it.
  • the height variation obtained by adding the height of the protrusion to the height of the base is controlled within a range of 20 ⁇ m.
  • a sampling device including the micro sample piece collecting tool according to the above [1] to [5], which has a holding portion for holding the micro sample piece collecting tool, and has a protrusion on the object to be analyzed. It has a drive mechanism that moves the micro sample piece collecting tool to a position where the portion is pierced or can be pressed, and moves the micro sample piece collecting tool along the object to be analyzed, and is minute. At the time of collecting the sample piece, the holding portion of the minute sample piece collecting tool is arranged at a distant position where the object to be analyzed does not come into contact.
  • a minute sample piece from various kinds of objects to be analyzed, not limited to living tissues, and in the process of collecting the minute sample piece from the object to be analyzed, a contaminant substance becomes a minute sample piece. It is possible to eliminate contamination and provide a minute sample piece in a state suitable for analysis.
  • FIG. 1 It is explanatory drawing which shows an example of the situation which the micro sample piece S is collected by the micro sample piece collecting tool 10. It is a figure which shows the micro sample taking chip 10A which concerns on 1st example, and is the figure which shows the example which the protrusion 12A was composed of a cone or a truncated cone. It is a top view which shows the example which arranged the protrusion 12A by increasing the arrangement density with respect to the example shown in FIG. 2 (a). It is a top view which shows the example which the protrusion 12A was gathered in the central part of the base part 11.
  • FIG. 10C It is a top view which shows the example which changed the arrangement of the protrusion 12C with respect to the micro sample piece collecting tool 10C shown in FIG. It is a top view which shows the example of the micro sample piece collecting tool 10C which gathered the protrusion 12C in the central part of the base 11. It is a figure which shows the micro sample taking chip 10D which concerns on 4th example, and is the figure which shows the example which the protrusion 12 was formed into a trapezoid body. It is a figure which shows the micro sample taking chip 10E which concerns on 5th example, and is the figure of the example in which the protrusion 12E is formed by a cylinder.
  • FIGS. 1 to 17 are schematic diagram showing the actual shape and configuration in a simplified manner.
  • FIG. 1 is an explanatory diagram showing an example of a situation in which a minute sample piece S (see FIG. 16) is collected by the minute sample piece collecting tool 10.
  • the micro sample piece collecting tool 10 is a micro device made of silicon, which is an inorganic solid material, formed of, for example, a silicon substrate, and has a rectangular planar shape.
  • the micro sample piece collecting tool 10 is formed with a plurality of protrusions 12 projecting to one surface side of the base 11 integrally with the base 11.
  • the protrusion 12 is a portion formed by processing a silicon substrate using a photolithography technique. In other words, the protrusion 12 is a portion that is surrounded by the etch groove and projects upward from the upper surface (one surface) of the base 11 that is the bottom of the etch groove.
  • the micro sample piece collecting tool 10 can be manufactured by one batch processing, for example, by forming a group of a large number of protrusions 12 on a 6-inch wafer and then individualizing the small sample piece by dicing
  • the material of the minute sample piece collecting tool 10 glass, ceramic, metal, or the like can be adopted in addition to silicon. However, it is a material that has excellent mechanical strength and is chemically stable, has no effect even if it comes into contact with the object to be analyzed, does not generate contaminants during the sampling process, and further. It is preferable to select silicon because the collected minute sample piece S can be easily separated from the minute sample piece collecting tool 10.
  • the total thickness of the silicon substrate is 640 ⁇ m
  • the height of the protrusion 12 is 10 ⁇ m
  • the plane size is a square of 2 mm ⁇ 2 mm.
  • these shapes and dimensions are only examples, and can be arbitrarily changed depending on the type, state, thickness, collection amount, and the like of the object to be analyzed. It is preferable that the height variation including the base 11 of the plurality of protrusions 12 is controlled so as to be within the range of 20 ⁇ m.
  • the object to be analyzed can be an organic substance or an inorganic substance and can be applied to any viscoelastic body.
  • the organic matter tissues such as humans and animals and plants other than humans, organic tissues attached to articles, and organic substances such as paste-like or solidified oil can be collected.
  • the micro sample piece collecting tool 10 can handle inorganic substances as long as the object to be analyzed is a viscoelastic body.
  • the collected micro sample piece can be used for genome analysis if the object to be analyzed is a biological tissue or the like, and can be used for elemental analysis if the object to be analyzed is another organic substance or inorganic substance.
  • the protrusion 12 As a method of collecting the minute sample piece S, the protrusion 12 is pierced or pressed against the thin section 14 of the object to be analyzed attached to the slide glass 13, and the protrusion 12 is relative to the thin section 14. Move to or turn. By doing so, the fine sample piece S of the viscoelastic body adheres to the protrusion 12. After that, if the micro sample piece S is immersed in the separation liquid or the like with the micro sample piece S attached to the micro sample piece collecting tool 10, the micro sample piece S can be separated from the micro sample piece collecting tool 10.
  • the thin section 14 may be prepared by slicing a mass of an object to be analyzed, for example.
  • each form of the micro sample piece collecting tool may be collectively referred to as a micro sample piece collecting tool 10
  • each form of the protrusion may be collectively referred to as a protrusion 12.
  • FIG. 2 is a diagram showing a micro sample piece collecting tool 10A according to the first example, and is a diagram showing an example in which the protrusion 12A is composed of a cone or a truncated cone.
  • FIG. 2A is a perspective view showing the micro sample piece collecting tool 10A
  • FIG. 2B is an enlarged plan view showing the protrusion 12A
  • FIG. 2C is a side view showing the protrusion 12A.
  • 2 (d) is a side view showing a case where the protrusion 12A is a conical body.
  • the micro sample piece collecting tool 10A has a plurality of protrusions 12A on one surface side of the base 11.
  • the protrusions 12A are arranged in a grid pattern extending in the X and Y directions.
  • the protrusion 12A shown in FIGS. 2 (a), 2 (b), and 2 (c) is a truncated cone having a top surface 16 such that the top of the cone is cut by a plane parallel to the bottom surface 15.
  • the area of the top surface 16, the area of the bottom surface 15, and the height of the top surface 16 from the base 11 can be arbitrarily set depending on the type and state of the object to be analyzed and the etching conditions. It is also possible to bend the side surface 17 of the protrusion 12A in the vertical direction.
  • the protrusion 12A is a cone having a vertex 18.
  • the microsample piece collecting tool 10A is moved in one direction in the X direction or the Y direction, reciprocated, or moved in a state where the apex 18 is pierced into the object to be analyzed. It is possible to collect the minute sample piece S (see FIG. 16) by scraping it by moving it diagonally with respect to the axis or further turning it.
  • the minute sample piece S is moved by moving the minute sample piece collecting tool 10A with respect to the object to be analyzed while the top surface 16 is pressed against the object to be analyzed. It becomes possible to collect by scraping.
  • the protrusion 12A can similarly collect a minute sample piece S in either a conical body or a truncated cone.
  • the protrusion 12A is a cone, it is possible to pierce the apex 18 into the object to be analyzed and collect the minute sample piece S, but it is conceivable that the top is missing.
  • the protrusion 12A by making the protrusion 12A a truncated cone, the strength of the top can be increased and the occurrence of chipping can be prevented. Since the micro sample piece S is a viscoelastic body, the micro sample piece S adheres to the periphery of the protrusion 12A.
  • the arrangement density and arrangement method of the protrusions 12A are not limited to the example shown in FIG. 2A, and can be freely set. An example thereof will be described with reference to FIGS. 3 and 4. Note that FIGS. 3 and 4 show an example in which the shape of the protrusion 12A is a truncated cone.
  • FIG. 3 is a plan view showing an example in which the protrusions 12A are arranged with a higher arrangement density than the example shown in FIG. 2 (a).
  • the number of protrusions 12A arranged on the base 11 is 13 ⁇ 13 in the X direction and the Y direction in the example shown in FIG. 2 (a), but in the example shown in FIG. 3, the number is 13 ⁇ 13 in the X direction and the Y direction.
  • 24 x 24 pieces are arranged in each. That is, the arrangement density of the protrusions 12A is about 3.4 times. By increasing the arrangement density in this way, the minute sample piece S can be subdivided and collected.
  • FIG. 4 is a plan view showing an example of a micro sample piece collecting tool 10A in which the protrusions 12A are assembled in the central portion of the base 11.
  • FIG. 4 is a diagram showing an example in which the protrusions 12A are arranged in seven rows in each of the X direction and the Y direction, and the arrangement density is sparser than in the example shown in FIG.
  • the collection function of each of the protrusions 12A is the same as the example shown in FIGS. 2 and 3, but the arrangement density depends on the plane size of the collection target portion of the object to be analyzed or the collection amount of the minute sample piece S. Can be selected as appropriate. It is possible to change the bottom surface area and the inclination angle of the side surface according to the arrangement density of the protrusion 12A.
  • FIG. 5 is a diagram showing a micro sample piece collecting tool 10B according to the second example, and is a diagram showing an example in which the protrusion 12B is composed of a quadrangular pyramid or a quadrangular pyramid.
  • 5 (a) is a perspective view showing a micro sample piece collecting tool 10B
  • FIG. 5 (b) is a plan view showing an enlarged protrusion 12B
  • FIG. 5 (c) is a side view
  • FIG. 5 (d) is. Is a side view showing the case where the protrusion 12B is a quadrangular pyramid.
  • the micro sample piece collecting tool 10B has a plurality of protrusions 12B on one surface side of the base 11.
  • the protrusions 12B are arranged in a grid pattern extending in the X and Y directions.
  • the protrusion 12B shown in FIGS. 5 (a), 5 (b), and 5 (c) is a quadrangular pyramid having a top surface 21 such that the top of the quadrangular pyramid is cut by a plane parallel to the bottom surface 20. It is a stand.
  • the area of the top surface 21, the area of the bottom surface 20, and the height of the top surface 21 from the base 11 can be arbitrarily set depending on the type and state of the object to be analyzed and the etching conditions.
  • the example shown in FIG. 5D is a quadrangular pyramid in which the protrusion 12B has the apex 22.
  • the minute sample piece collecting tool 10B may be moved in one direction in the X or Y direction or reciprocated while the apex 22 is pierced into the object to be analyzed. It is possible to collect the minute sample piece S (see FIG. 16) by scraping it by moving it diagonally with respect to the X-axis or further turning it.
  • the protrusion 12B When the protrusion 12B is a quadrangular pyramid, the micro sample piece S is scraped off by moving the micro sample piece collecting tool 10B while the top surface 21 is pressed against the object to be analyzed. It becomes possible to do.
  • the protrusion 12B can similarly collect a minute sample piece S from either a quadrangular pyramid or a quadrangular frustum.
  • the protrusion 12B When the protrusion 12B is a cone, it is possible to pierce the apex 22 into the object to be analyzed and collect the minute sample piece S, but it is conceivable that the top is missing.
  • the protrusion 12B by making the protrusion 12B a quadrangular pyramid, the strength of the top can be increased and the occurrence of chipping can be prevented.
  • the collection status of the minute sample piece S differs depending on the moving direction of the minute sample piece collecting tool 10B. .. For example, if the minute sample piece collecting tool 10B is moved in the X direction or the Y direction, the minute sample piece S can be collected so as to be scraped off. If the minute sample piece collecting tool 10B is moved obliquely with respect to the X-axis or the Y-axis, the minute sample piece S can be collected by scraping off the object to be analyzed by the ridge line 23. Since the micro sample piece S is a viscoelastic body, the micro sample piece S adheres to the periphery of the protrusion 12B.
  • the arrangement density and arrangement method of the protrusion 12B are not limited to the example shown in FIG. 5A, and can be freely set.
  • the protrusions 12B may be assembled at the center of the base 11, or the protrusions 12B may be rotated 90 degrees with respect to the placement angle (posture) of the protrusions 12B shown in FIG. 5A. Is possible.
  • FIG. 6 is a plan view showing an example of a micro sample piece collecting tool 10B in which the protrusions 12B are assembled in the central portion of the base 11.
  • FIG. 6 shows an example in which the protrusions 12B are arranged in six rows in each of the X direction and the Y direction.
  • the collection function of each of the protrusions 12B is the same as the example shown in FIG. 5, but depending on the plane size of the collection target portion of the object to be analyzed, or the collection amount of the minute sample piece S (see FIG. 16). ,
  • the number of arrangements and the arrangement layout of the protrusions 12B can be appropriately selected. It is possible to change the area of the bottom surface and the inclination angle of the side surface according to the arrangement density of the protrusion 12B.
  • FIG. 7 is a plan view showing a micro sample piece collecting tool 10B in which the arrangement angle (posture) of the protrusion 12B is changed with respect to the protrusion 12B shown in FIG.
  • the protrusion 12B shown in FIG. 7 is formed by rotating the protrusion 12B by 90 degrees about the central axis 24 which passes through the apex 22 and is perpendicular to the bottom surface 20 in FIG.
  • the protrusions 12B By arranging the protrusions 12B in this way, when the minute sample piece collecting tool 10B is moved in the X direction or the Y direction, the minute sample piece S is scraped off while scraping the object to be analyzed along the ridge line 23. It becomes possible to collect.
  • the arrangement angle of the protrusion 12B can be appropriately set.
  • the arrangement angle of the protrusions 12B may be the same as the arrangement angle of the protrusions 12B shown in FIG. It is possible, and the arrangement angle of the protrusion 12B can be arbitrarily set.
  • FIG. 8 is a diagram showing the micro sample piece collecting tool 10C according to the third example, and is a diagram showing an example in which the protrusion 12C is composed of a triangular pyramid or a triangular pyramid.
  • 8 (a) is a perspective view showing the minute sample piece collecting tool 10C
  • FIG. 8 (b) is an enlarged plan view of the protrusion 12C
  • FIG. 8 (c) is a side view
  • FIG. 8 (d) is. Is a side view showing the case where the protrusion 12C is a triangular pyramid.
  • the micro sample piece collecting tool 10C has a plurality of protrusions 12C on one surface side of the base 11.
  • the protrusion 12C is arranged so that one of the three ridges, the ridge 26, faces the direction orthogonal to the side 11a of the base 11.
  • the protrusion 12C shown in FIGS. 8 (a), 8 (b), and 8 (c) is a triangular pyramid having a top surface 28 such that the top of the triangular pyramid is cut by a plane parallel to the bottom surface 27. It is a stand.
  • the protrusion 12C shows an example in which the shape of the bottom surface 27 is an equilateral triangle, but the shape of the bottom surface 27 is not limited to an equilateral triangle.
  • the area of the top surface 28, the area of the bottom surface 27, and the height of the top surface 28 can be arbitrarily set according to the type and state of the object to be analyzed and the etching conditions.
  • the example shown in FIG. 8D is a triangular pyramid in which the protrusion 12C has an apex 29.
  • the protrusions 12C are arranged in a grid pattern extending in the X and Y directions. Since the protrusion 12C has ridges 26 on three sides, the collection status of the minute sample piece S differs depending on the moving direction with respect to the object to be analyzed. For example, if the minute sample piece collecting tool 10C is moved in the extension direction of the ridge line 26, the minute sample piece S can be collected by scraping the minute sample piece S while being scraped off by the ridge line 26. If it moves in the opposite direction, the minute sample piece S can be collected so as to be scraped off. As shown in FIG.
  • the object to be analyzed when the protrusion 12C is a cone, the object to be analyzed is pierced with the apex 29, and the object to be analyzed is unidirectional in the X direction or the Y direction. It is possible to collect the minute sample piece S by scraping it by moving it to, reciprocating, moving it diagonally with respect to the X-axis, or even turning it.
  • the protrusion 12C When the protrusion 12C is a triangular frustum, the micro sample piece S is scraped off by moving the micro sample piece collecting tool 10C while the top surface 28 is pressed against the object to be analyzed. It becomes possible to do.
  • the protrusion 12A can similarly collect a minute sample piece S in either a conical body or a truncated cone.
  • the protrusion 12C When the protrusion 12C is a cone, it is possible to pierce the apex 18 into the object to be analyzed and collect the minute sample piece S, but it is conceivable that the top is missing.
  • the protruding portion 12C as a triangular frustum, the strength of the top portion can be increased and the occurrence of chipping can be prevented. Since the micro sample piece S is a viscoelastic body, the micro sample piece S adheres to the periphery of the protrusion 12C.
  • FIG. 9 is a plan view showing an example in which the arrangement of the protrusions 12C is changed with respect to the micro sample piece collecting tool 10C shown in FIG.
  • the protrusions 12C are arranged by changing the arrangement angle (posture) for each row in the X direction.
  • the protrusion 12C-1 arranged in the first row on the leftmost side is arranged so that one extension line of the ridge line 26 faces the direction orthogonal to the side 11a in the X direction, and is arranged in the second row from the left side.
  • the protrusions 12C-2 are arranged so that one of the sides of the bottom surface 27 is parallel to the side 11a and further offset by half a pitch with respect to the first row.
  • One protrusion 12C-2 is arranged at the center of four protrusions 12C-1.
  • the respective protrusions 12C-1 and 12C-2 By arranging the protrusions 12C-1 and 12C-2 in this way, even if the minute sample piece collecting tool 12C is moved or swiveled in the X direction, the Y direction, or an arbitrary direction, the respective protrusions have the same conditions. It becomes possible to collect a minute sample piece S.
  • FIG. 10 is a plan view showing an example of a micro sample piece collecting tool 10C in which the protrusions 12C are assembled in the central portion of the base 11.
  • the protrusions 12C have their respective plane postures set (the postures do not have to be set under special rules), and are arranged in a substantially circular shape.
  • each protrusion arranged on the concentric circles is arranged so as to form a predetermined angle with respect to the circumference.
  • the collection function of each of the protrusions 12C is the same, but the posture and arrangement of the protrusions 12C can be appropriately selected depending on the plane size of the collection target portion of the object to be analyzed or the collection amount of the minute sample piece S. It is possible.
  • FIG. 11 is a diagram showing the micro sample piece collecting tool 10D according to the fourth example, and is a diagram showing an example in which the protrusion 12D is formed of a trapezoid body.
  • 11 (a) is a perspective view of the micro sample piece collecting tool 10D
  • FIG. 11 (b) is an enlarged perspective view of one of the protrusions 12D
  • FIG. 11 (c) is a modified example of the fourth example.
  • the micro sample piece collecting tool 10D has a plurality of protrusions 12D on one surface side of the base 11.
  • the bottom surface 32 and the top surface 33 of the protrusion 12D are rectangular, and the top surface 33 is parallel to the bottom surface 32.
  • the protrusions 12D are arranged so that the long side 34 of the bottom surface 32 is parallel to the side 11a of the base 11.
  • the shape of the protrusion 12D is arbitrarily set according to the type and state of the object to be analyzed and the etching conditions, such as the area of the top surface 33, the area of the bottom surface 33, the height of the top surface 33 from the base 11, and the inclination angle of the side surface. It is possible to do. As shown in FIG. 11 (c), it is also possible to extend the four ridge lines 35 to have a straight portion 36 connecting the intersections of the two sets of ridge lines.
  • the micro sample piece collecting tool 10D shown in FIG. 11 has a shape suitable for collecting the micro sample piece S by moving it in the X direction.
  • the fourth example it is possible to increase the arrangement density of the protrusions 12D and to gather the protrusions 12D at the center of the base 11.
  • a row extending in the Y direction it is possible to widen the gap between the protrusions 12D so that the protrusions 12D in the adjacent row are arranged between the gaps, and the arrangement angle (posture) of the protrusions 12D is possible.
  • the minute sample piece S is collected by moving the minute sample piece collecting tool 10D in the X direction so that the object to be analyzed is scraped off by the corner portion or the straight portion 36 of the top surface 33. It becomes possible to do.
  • FIG. 12 is a diagram showing the micro sample piece collecting tool 10E according to the fifth example, and is a diagram showing an example in which the protrusion 12E is formed of a cylinder.
  • 12 (a) is a perspective view showing the micro sample piece collecting tool 10E
  • FIG. 2 (b) is a side view showing the protrusion 12E
  • FIG. 2 (c) is an example in which the protrusion 12E has an inverted tapered shape. It is a side view which shows.
  • the protrusions 12E are arranged in a grid pattern extending in the X and Y directions.
  • FIG. 12C has a tapered shape in which the area of the bottom surface 38 of the protrusion 12E is smaller than the area of the top surface 37. It has a so-called reverse taper shape.
  • the micro sample piece collecting tool 10E the micro sample piece S is collected by pressing the top surface 37 against the object to be analyzed and moving the sample piece S so as to be scraped off by the corner between the top surface 37 and the side surface 39.
  • the angle formed by the top surface 37 and the side surface 39 is an acute angle as compared with the case of a simple cylinder. It is possible to increase the ability of the collecting tool 10E to collect minute pieces.
  • the second row of protrusions 11E may be arranged with a half pitch shift in the Y direction with respect to the arrangement of the protrusions 11E on the side 11a side of the base 11. That is, it is possible to change the arrangement of the protrusions 12E for each row in the X direction.
  • FIG. 13 is a diagram showing a micro sample piece collecting tool 10F according to the sixth example, and is a diagram showing an example in which the protrusion 12F is composed of a quadrangular prism.
  • the micro sample piece collecting tool 10F has a plurality of protrusions 12F on one surface side of the base 11.
  • the protrusions 12F are arranged in a grid pattern extending in the X and Y directions.
  • the protrusion 12F is a regular tetrahedron having a square top surface 41, but the top surface 41 may be rectangular.
  • the protrusion 12F is arranged so that one of the four ridge lines 40 (lines constituting the height) faces the side 11a of the base portion 11.
  • the area of the top surface 41 of the protrusion 12F and the height from the base 11 can be arbitrarily set according to the type and state of the object to be analyzed and the etching conditions.
  • the minute sample piece collecting tool 10F If the minute sample piece collecting tool 10F is moved in the X direction or the Y direction, the minute sample piece S can be collected by scraping the minute sample piece while scraping it along the ridge line 40.
  • the micro sample piece collecting tool 10F moves, reciprocates, moves diagonally with respect to the X axis, and further rotates the object to be analyzed in one direction of the X direction or the Y direction. This makes it possible to collect the minute sample piece S.
  • the arrangement of the protrusions 12F is not limited to the example shown in FIG.
  • the second row of protrusions 12F may be arranged with a half pitch shift in the Y direction with respect to the arrangement of the protrusions 12F on the side 11a side of the base 11. It is also possible to arrange one side of the bottom surface of the second row of protrusions 12F shifted by half a pitch so as to be parallel to the side 11a of the base portion 11. That is, it is possible to change the arrangement angle (posture) of the protrusions 12E for each row in the X direction.
  • the side surface 42 of the protrusion 12F can also have a reverse taper shape as in the example shown in FIG. 12 (a).
  • the angle formed by the top surface 41 and the side surface 42 becomes an acute angle as compared with the case of a simple quadrangular prism. It becomes possible to increase.
  • FIG. 14 is a diagram showing a micro sample piece collecting tool 10G according to the seventh example, and is a diagram showing an example in which the protrusion 12G is composed of a triangular prism.
  • the micro sample piece collecting tool 10G has a plurality of protrusions 12G on one surface side of the base 11.
  • the protrusions 12G are arranged in a grid pattern extending in the X and Y directions.
  • the protrusion 12G is an equilateral triangular prism whose top surface 45 is an equilateral triangle, but the top surface shape is not limited to an equilateral triangle.
  • the protrusion 12G shown in FIG. 14 is arranged so that one side of the top surface 45 is parallel to the side 11a of the base portion 11.
  • the area of the top surface 45 and the height from the base 11 can be arbitrarily set according to the type and state of the object to be analyzed and the etching conditions.
  • the micro sample piece collecting tool 10G moves, reciprocates, moves diagonally with respect to the X axis, and further rotates the object to be analyzed in one direction in the X or Y direction. It becomes possible to collect a minute sample piece S.
  • the arrangement of the protrusions 12G is not limited to the example shown in FIG.
  • the protrusions 12G in the second row may be arranged with a half pitch shift in the Y direction with respect to the arrangement of the protrusions 12G in the first row from the side 11a of the base 11. It is also possible to rotate the protrusion 12G in the second row, which has even a half pitch, by 180 degrees with respect to the first row. That is, it is possible to change the arrangement angle (posture) of the protrusions 12G for each row in the X direction.
  • the side surface 46 of the protrusion 12G can have an inverted tapered shape as in the example shown in FIG. 12 (a).
  • the angle formed by the top surface 45 and the side surface 46 becomes an acute angle as compared with the case of a simple triangular prism. It becomes possible to increase.
  • the micro sample piece collecting tools 10A to 10G described above have protrusions 12A to 12G having shapes such as a cone, a thruster, and a pillar arranged on one surface side of the base 11, but the shape of the protrusions. And arrangement are not limited to this, and can be set arbitrarily. For example, it is possible to mix a plurality of types of protrusions on the base 11. Further, the protrusions 12A to 12G can be formed on both the front and back surfaces of the base portion 11, and the configurations of the protrusions 12A to 12G can be changed on the front and back surfaces.
  • the micro sample piece collecting tools 10A to 10G are manufactured by processing a silicon substrate by using photolithography technology, the protrusions 12A to 12G are formed for each micro sample piece collecting tool forming region of the silicon substrate. It is also possible to form one of them and prepare a plurality of types of microsample collection tools by dicing. Since the total thickness of the micro sample piece collecting tool 10A to 10G including the protrusions corresponds to the total thickness of the silicon substrate, the heights of the protrusions 12A to 12G for each silicon substrate from the top surfaces and the bases 11 of the vertices. It is possible to keep the variation of the sample small.
  • micro sample piece collecting tools 10A to 10G exemplified in the first to seventh examples form each protrusion by photoetching, which is a part of the semiconductor manufacturing process, they are analyzed during the forming process. It is a clean microdevice that does not contain contaminants that affect the analysis of objects.
  • the micro sample piece collecting tools 10A to 10G can collect the micro sample piece S (see FIG. 16) without being limited to an organic substance or an inorganic substance as long as the object to be analyzed is a viscoelastic body.
  • the object to be analyzed is a pathological tissue sample for genomic medicine, and as a specific example, the minute sample piece S is collected from the object to be analyzed using the minute sample piece collecting tool 10A.
  • FIG. 15 is an explanatory diagram showing an example of a situation in which the micro sample piece S is collected from the thin section 14 of the object to be analyzed by the micro sample piece collecting tool 10A.
  • the thin section 14 is a tissue obtained by surgery or the like, which is subjected to paraffin embedding treatment and then sliced to a thickness of 3 ⁇ m to 4 ⁇ m.
  • the thin section 14 is attached to a slide glass 13 after identifying a tissue collection target area (for example, a cancer tissue area) by HE staining.
  • a tissue collection target area for example, a cancer tissue area
  • the micro sample piece S is scraped off by moving or swirling along the thin section 14 in a state where the protrusion 12A is pierced or pressed against the collection target area of the thin section 14.
  • the fine sample piece S of the viscoelastic body adheres to the protrusion 12A and the periphery of the protrusion 12A.
  • the arrows in the figure indicate the moving direction of the minute sample piece collecting tool 10A.
  • the thin section 14 may be moved by fixing the micro sample piece collecting tool 10A.
  • the micro sample piece collecting tool 10A is a micro device having a side surface size of 2 mm ⁇ 2 mm and a thickness of 640 ⁇ m
  • the micro sample piece collecting tool 10A is attached to the holder 50 to collect the micro sample piece S.
  • the micro sample piece collecting tool 10A is removable from the holder 50. Since the micro sample piece S may adhere to the holder 50 when the micro sample piece S is collected, it is preferable that the holder 50 is discarded together with the micro sample piece collecting tool 10A. That is, it is preferable that the micro sample piece collecting tool 10A and the holder 50 are disposable. When collecting a small sample piece, it is also possible to grasp the holder 50 by hand and perform the collection operation.
  • the minute sample piece S to the driving means via the holder 50 and automatically or semi-automatically drive the minute sample piece collecting tool 1 to collect the minute sample piece S. It is also possible to collect the minute sample piece collecting tool 10 using tweezers, a clamp, or the like. This sampling operation is performed in the chamber so that contaminants do not adhere to the minute sample piece S.
  • FIG. 16 is a micrograph showing a state after collecting the minute sample piece S with the minute sample piece collecting tool 10A.
  • a minute sample piece S scraped from the thin section 14 is attached to the periphery of the protrusion 12A.
  • an average of 0.05 mg of the micro sample piece S could be collected per micro sample piece collecting tool. Since the amount of DNA required for genomic medicine is 0.01 mg or more, a sufficient amount can be obtained by one small sample piece collecting tool and one collecting operation.
  • the collected micro sample piece S is irregularly miniaturized, it can be analyzed as a pathological tissue sample for genomic medicine.
  • FIG. 17 is an explanatory diagram showing an example of a schematic configuration of the sampling device 55.
  • the collecting device 55 holds the stage 56 on which the slide glass 13 is placed and the micro sample piece collecting tool 10A, and the micro sample piece collecting tool 10A is used as the object to be analyzed, and the protrusion 12A is provided.
  • It has a drive mechanism 57 as a drive means for rotating and turning in the X direction, the Y direction, the Z direction, and in a pierced state or a pressed state.
  • the micro sample piece collecting tool 10A is attached to the connecting portion 58, which is a part of the driving mechanism 57, via the holder 50.
  • the holder 50 is removable from the connecting mechanism 58.
  • the micro sample piece collecting tool 10A can be directly attached to the connecting portion 58, and the micro sample piece collecting tool 10 has a structure that is detachable from the connecting portion 58.
  • the slide glass 13 to which the thin section 14 is attached is placed on the stage 56 and fixed so as not to move when the minute sample piece S is collected.
  • the drive mechanism 57 lowers the micro sample piece collecting tool 10A to a position where the protrusion 12A can collect the micro sample piece S from the thin section 14, and keeps the height position of the micro sample piece collecting tool 10A. It is moved in the X and Y directions along the thin section 14, or rotated or swiveled at that position.
  • the micro sample piece collecting tool 10A is raised, the micro sample piece collecting tool 10A is separated from the holder 50, and the holder 50 is separated from the connecting portion 58.
  • the minute sample piece S adheres to the minute sample piece collecting tool 12A in the state as described with reference to FIG.
  • the micro sample piece S is collected from the thin section 14 prepared by slicing the mass of the object to be analyzed, but the micro sample piece is directly collected from the object to be analyzed in the state of the mass. It is also possible to collect.
  • the minute sample piece S can be collected by appropriately adjusting the distance of the minute sample piece collecting tool 10 from the object to be analyzed and appropriately selecting the shape including the height of the protrusion 12. Become.
  • the stage is provided with a drive mechanism, and the thin section 14 can be moved with respect to the minute sample piece collecting tool 10A.
  • the micro sample piece collecting tool 10 described above is a micro sample piece collecting tool for collecting a part of the object to be analyzed as a micro sample piece S.
  • the micro sample piece collecting tool 10 is formed integrally with the base portion 11 made of an inorganic solid material and the base portion 11, and protrudes to one surface side of the base portion 11 so that the tip portion is pierced into the object to be analyzed of the viscoelastic body. Alternatively, it has a protrusion 12 that scrapes off a part of the object to be analyzed by moving relative to the object to be analyzed in a pressed state.
  • the sampling device described in Patent Document 1 and Patent Document 2 described above is limited to the sampling of thin sections of biological tissue to be sampled, and dust, abrasion powder, and processing associated with the manufacture and operation of the sampling device. Since there are many factors that can be pollutants such as oil, and multiple steps are required to collect micro sample pieces, there are many opportunities for pollutants to be mixed in the object to be analyzed, and the micro is suitable for analysis. There was a risk that the sample piece could not be provided.
  • the micro sample piece collecting tool 10 (micro sample piece collecting tool 10A to 10G) according to the present embodiment is formed of an inorganic solid material, and the protrusion 12 (protrusion 12A to 12G) is pierced into the object to be analyzed.
  • the small sample piece S is collected by scraping it off with the protrusion 12 by pressing it against the surface and moving it. Therefore, even if the object to be analyzed is an organic substance or an inorganic substance, if it is a viscoelastic body, the micro sample piece S can be collected by the micro sample piece collecting tool 10.
  • the micro sample piece collecting tool 10 it is possible to collect the micro sample piece S not only from the living tissue but also from various objects to be analyzed, and the pollutants are collected in the process of collecting the micro sample piece S. It is possible to provide a minute sample piece S in a state suitable for analysis by eliminating the fact that is mixed in the object to be analyzed.
  • the inorganic solid material is a silicon substrate, and the protrusions 12A to 12G are formed by using photolithography technology.
  • the protrusions 12A to 12D are portions that are surrounded by the etch groove and project upward from the upper surface (one surface) of the base portion 11 that is the bottom of the etch groove.
  • Silicon is a material that has excellent mechanical strength and is chemically stable, and does not have any effect even if it comes into contact with the object to be analyzed. Further, since the micro sample piece collecting tools 10A to 10G are formed by using photolithography technology (implemented in a clean environment with advanced automation) which is a part of the semiconductor manufacturing process, the structure constitutes the mechanism. In the process of manufacturing parts and assembling the sampling device, dust, dust due to wear of parts, processing oil, worker's oil and fat, and body fluid do not adhere or remain. Further, there is an effect that the minute sample piece S can be easily separated from the minute sample piece collecting tools 10A to 10G. Further, the shape and height of the protrusion 12 can be easily controlled by selecting the thickness of the silicon substrate or changing the etching conditions.
  • the protrusions 12A to 12C are composed of a cone, a thruster, or a pillar.
  • the protrusion shape is a cone
  • the top is pierced into the object to be analyzed, and in the case of a thruster or a prism, the top surface is pressed against the object to be analyzed and moved to move the minute sample piece S. Can be collected by scraping.
  • the shape of the protrusion is a cone, it is possible that the top is chipped, but the strength can be increased by using a thruster or a prism.
  • the protrusions 12E, 12F, and 12G are composed of pillars.
  • the protrusions 12E, 12F, and 12G have a tapered shape in which the area of the bottom surface of the prism is smaller than the area of the top surface.
  • the relationship is "the area of the bottom surface 38 ⁇ the area of the top surface 37". That is, the protrusion 12E has an inverted tapered shape. Even if the protrusions are square prisms and triangular prisms, it is possible to form an inverted taper shape in the same manner as a cylinder.
  • the angle between the top surface and the side surface becomes acuteer than in the case of a simple prism, so that the ability to collect minute sample pieces is enhanced. Is possible.
  • the variation in height obtained by adding the height of the protrusion to the thickness of the base 11 is controlled within a range of 20 ⁇ m.
  • each minute sample piece collecting tool keeps the height position of the protrusion within a certain range, so that the amount and collection of the minute sample piece S in each protrusion are collected. It is possible to suppress variations in the situation. Since the micro sample piece collecting tools 12A to 12G are created from a silicon substrate by photolithography technology, the apex or top surface of each of the protrusions 12A to 12G corresponds to one surface of the silicon substrate. , The variation in height can be suppressed to an extremely small size.
  • the sampling device 55 has a holding portion for holding the micro sample piece collecting tool 10, moves to a position where the protrusion 12 is pierced or pressed against the object to be analyzed, and is a micro sample. It has a drive mechanism 57 that moves the piece collecting tool 10 along the object to be analyzed, and when collecting the minute sample piece, the holding portion (holder 50 or the connecting part 58) of the minute sample piece collecting tool 10 is from the object to be analyzed. It is located at a distance.
  • sampling device 55 By using such a sampling device 55, it becomes possible to appropriately control the load of pressing the minute sample piece collecting tools 10A to 10G against the object to be analyzed, the moving direction and the moving speed at the time of sampling, and it is constant. It is possible to efficiently collect the minute sample piece S under the conditions. In the sampling work by such a sampling device 55, if a clean micro sample piece collecting tool 10 is attached, the sampling work can be performed immediately. It is possible to provide a minute sample piece in a state suitable for analysis without being mixed with the analysis target.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

La présente invention concerne un outil de collecte de micro-échantillon 10 qui comprend : une partie de base 11 constituée d'un matériau individuel inorganique; et des saillies 12 qui sont formées d'un seul tenant avec la partie de base 11, font saillie depuis un côté de surface de la partie de base 11, et raclent une partie d'un objet à analyser d'un corps viscoélastique en se déplaçant par rapport à l'objet à analyser dans un état dans lequel une partie de pointe est percée ou pressée contre l'objet à analyser. Un dispositif de collecte 55 comprend l'outil de collecte de micro-échantillon 10, un support 50 qui maintient l'outil de collecte de micro-échantillon 10, et un mécanisme d'entraînement 57 qui déplace les saillies vers une position dans laquelle les saillies percent ou pressent contre l'objet à analyser, et déplace les saillies 12 le long de l'objet à analyser. Cela permet de collecter des micro-échantillons à partir de différents objets à analyser dans un état adapté pour l'analyse sans mélange avec des contaminants.
PCT/JP2021/008896 2020-04-02 2021-03-08 Outil de collecte de micro-échantillon et dispositif de collecte WO2021199926A1 (fr)

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JP2020-066869 2020-04-02
JP2020066869A JP2021162533A (ja) 2020-04-02 2020-04-02 微小試料片採取具及び採取装置

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006061991A (ja) * 2004-08-24 2006-03-09 Canon Inc サンプリング用治具及びその製造方法
US20120310113A1 (en) * 2008-12-01 2012-12-06 Oasis Diagnostics Corporation Biological sample collection system
WO2014038437A1 (fr) * 2012-09-04 2014-03-13 富士通株式会社 Montage d'échantillonnage, procédé d'analyse quantitative et système d'analyse
JP2017532549A (ja) * 2014-09-17 2017-11-02 ホロジック, インコーポレイテッドHologic, Inc. 分離可能な試料収集デバイス
CN206612814U (zh) * 2016-11-28 2017-11-07 广州毅得健生物科技有限公司 检体采检棒
JP2018512855A (ja) * 2015-04-08 2018-05-24 ベクトン・ディキンソン・アンド・カンパニーＢｅｃｔｏｎ， Ｄｉｃｋｉｎｓｏｎ Ａｎｄ Ｃｏｍｐａｎｙ 半固体表面から微生物増殖物を採取するデバイス及び装置

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006061991A (ja) * 2004-08-24 2006-03-09 Canon Inc サンプリング用治具及びその製造方法
US20120310113A1 (en) * 2008-12-01 2012-12-06 Oasis Diagnostics Corporation Biological sample collection system
WO2014038437A1 (fr) * 2012-09-04 2014-03-13 富士通株式会社 Montage d'échantillonnage, procédé d'analyse quantitative et système d'analyse
JP2017532549A (ja) * 2014-09-17 2017-11-02 ホロジック, インコーポレイテッドHologic, Inc. 分離可能な試料収集デバイス
JP2018512855A (ja) * 2015-04-08 2018-05-24 ベクトン・ディキンソン・アンド・カンパニーＢｅｃｔｏｎ， Ｄｉｃｋｉｎｓｏｎ Ａｎｄ Ｃｏｍｐａｎｙ 半固体表面から微生物増殖物を採取するデバイス及び装置
CN206612814U (zh) * 2016-11-28 2017-11-07 广州毅得健生物科技有限公司 检体采检棒

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