Filter Plate

Abstract

A filter plate (10) is attached to a sample container having a sample storage section with an open top, and comprises a base material (11, 31) having a first through-hole (112, 312) for passing a sample liquid at a position corresponding to the sample storage section, a first filter (12, 32) provided in the first through-hole, and a support (13, 33) arranged adjacent to the side of the first filter from which the sample liquid flows out and having a second through-hole larger than the pore size of the first filter. One embodiment of the filter plate (10) is used for a microplate having multiple wells, the base material is provided with the first through-hole at a position corresponding to each well of the microplate, and the first filter is a sheet-like filter arranged on one side of the base material so as to cover all of the first through-holes.

Description

[Technical field]

[0001] The present invention relates to a filter plate that is attached to a sample container such as a microplate to filter a sample liquid. [Background technology]

[0002] Conventionally, when a sample liquid derived from a living body or the like is injected into a well of a microplate, a filter is used to remove impurities from the sample liquid and extract a target substance from the sample liquid. The pore size of such a filter is small, and the sample liquid often does not pass through the filter when it is simply dropped onto the filter. For this reason, an operation (centrifugal filtration) is performed in which the sample liquid is dropped onto a filter provided corresponding to each well, and then the sample liquid is drawn into the well by applying centrifugal force using a centrifuge (e.g., Patent Documents 1 and 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Special Publication No. 2020-519440 [Patent Document 2] Special Publication No. 2007-509633 Summary of the Invention [Problem to be solved by the invention]

[0004] When centrifugal filtering of a sample liquid derived from a living organism, a thin filter with a small pore size is often used, and there is a problem that the filter is easily broken when subjected to operations that apply force to the filter, such as centrifugal filtration.

[0005] The problem that the present invention aims to solve is to provide a filter plate that can prevent the filter from being broken when performing an operation in which force is applied to the filter, such as centrifugal filtration of a sample liquid. [Means for solving the problem]

[0006] The present invention, which has been made to solve the above problems, provides a filter plate to be attached to a sample container having a sample storage portion with an open top, comprising: a) a substrate having a first through hole through which a sample liquid flows, the first through hole being provided at a position corresponding to the sample storage portion; b) a first filter provided in the first through hole; c) a support disposed adjacent to the first filter on the side from which the sample liquid flows out, the support having second through-holes formed therein and having a pore size larger than that of the first filter; The present invention is characterized by comprising:

[0007] The filter plate according to the present invention is fixed to a sample container so that the position of the opening of the sample container and the position of the first through-hole of the base material are aligned, and the support is located closer to the sample container than the filter. In the filter plate according to the present invention, a first filter used for centrifugal filtration or the like is provided in the first through-hole of the base material fixed to the sample container. The filter plate according to the present invention further includes a support disposed adjacent to the side of the first filter from which the sample liquid flows out. Therefore, when performing an operation in which force is applied to the filter, such as centrifugal filtration, the filter is supported from the back side (the side of the sample container), and damage to the first filter is suppressed. In addition, the support is formed with a second through-hole larger than the pore size of the first filter used for centrifugal filtration, so that the sample liquid that has passed through the filter is not prevented from being introduced into the sample container of the sample container.

[0008] In the filter plate according to the present invention, The filter plate is used for a microplate having a plurality of wells, and the first through-holes are provided in a base material at positions corresponding to each well of the microplate; The first filter is preferably a sheet-like filter disposed on one surface of the substrate so as to cover all of the first through-holes.

[0009] The filter plate according to the present invention can be used for a microplate having multiple wells, as in the above embodiment. In this case, the first filter can be easily attached by using a sheet-like filter disposed on one side of the base so as to cover all the first through-holes, as in the above embodiment.

[0010] In the filter plate according to the present invention, The support is preferably a second filter formed by laminating an adhesive sheet to a sheet-like material and forming the second through holes therein.

[0011] In the filter plate of the above embodiment, the first filter and the second filter are adhered to each other by the adhesive sheet of the support, thereby making it possible to prevent sample liquid from flowing between them.

[0012] The filter plate according to the present invention further comprises: a flexible, plate-like packing material that is disposed on the side of the support from which the sample liquid flows out and has third through-holes formed at positions corresponding to each of the plurality of wells; It is preferable to have

[0013] In the filter plate of the above embodiment, the flowing sample liquid can be introduced from the first through-hole of the substrate into the well corresponding to the through-hole without leakage. Effect of the Invention

[0014] By using the filter plate according to the present invention, damage to the filter can be suppressed when a force is applied to the filter, such as centrifugal filtration of a sample liquid. [Brief description of the drawings]

[0015] [Figure 1] 1A is a top view, FIG. 1B is a longitudinal sectional view, and FIG. 1C is a bottom view showing a first embodiment of a filter plate according to the present invention. [Diagram 2]3 is a longitudinal cross-sectional view showing a filter / packing seal including a filter and a packing, which is attached to a base material during the manufacture of the filter plate of the first embodiment. FIG. [Diagram 3] FIG. 2 is an exploded perspective view of the seal with filter and packing material in the first embodiment. [Figure 4] FIG. 2 is a longitudinal sectional view showing a state in which the filter plate of the first embodiment is attached to a microplate. [Diagram 5] FIG. 2 is a schematic diagram showing a state in which a microplate and a filter plate of the first embodiment are mounted on a centrifuge. [Figure 6] 5A is a top view, FIG. 5B is a longitudinal sectional view, and FIG. 5C is a bottom view showing a second embodiment of a filter plate according to the present invention. [Figure 7] 13A is a partially enlarged vertical cross-sectional view of a filter plate according to a second embodiment, and FIG. [Figure 8] FIG. 11 is a partially enlarged longitudinal sectional view showing a state in which a filter plate according to a second embodiment is attached to a microplate. [Figure 9] FIG. 6 is a partial vertical cross-sectional view showing a filter plate according to a modified example of the first embodiment. [Figure 10] FIG. 2 is a longitudinal sectional view of a filter plate according to an embodiment of the present invention. [Figure 11] A longitudinal cross-sectional view showing a filter / packing seal including a filter and a packing material, which is attached to a base material during the manufacture of a filter plate of a related embodiment. [Figure 12] FIG. 13 is an exploded perspective view of a seal with filter packing material in a related embodiment. [Figure 13] FIG. 11 is a partial enlarged view of a second filter sheet in a related embodiment. [Figure 14] FIG. 11 is a partially enlarged view showing how a pipette tip is inserted into one of the first through-holes and pressed against the second filter sheet to extract sample liquid in a related embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] An embodiment of a filter plate according to the present invention will be described with reference to FIGS.

[0017] (1) First embodiment Fig. 1 shows a filter plate 10 of the first embodiment. Fig. 1(a) is a top view of the filter plate 10, Fig. 1(b) is a vertical cross-sectional view of the filter plate 10 taken along line aa, and Fig. 1(c) is a bottom view of the filter plate 10. This filter plate 10 has a base material 11, a first filter sheet 12, a second filter sheet 13, and a packing material 14.

[0018] The base material 11 has a plastic plate-like member 111 having a rectangular planar shape, and a plurality of first through holes 112 having a circular planar shape provided in the plate-like member 111. A total of 96 first through holes 112 are provided, with 12 first through holes 112 arranged parallel to the long sides of the rectangle and 8 first through holes 112 arranged parallel to the short sides. The arrangement of the first through holes 112 corresponds to the arrangement of wells of a 96-hole microplate to which the filter plate 10 is attached. On the top surface of the base material 11, the end of the long side has numbers 1 to 12 written in accordance with the rows in which the first through holes 112 are arranged, and the end of the short side has eight letters A to H written in accordance with the rows in which the first through holes 112 are arranged. These numbers and letters are symbols for identifying each first through hole 112.

[0019] Of the four rectangular corners of the base material 11 (plate-shaped member 111), two adjacent corners across one of the short sides are cut off at an angle of 45° to the short side to form C-faces 113. By providing the C-faces 113, the orientation of the filter plate 10 can be easily recognized.

[0020] A rectangular frame-shaped surrounding wall 114 made of the same material as the base material 11 extends downward from the outer periphery of the lower surface of the base material 11. In this embodiment, the surrounding wall 114 is provided perpendicular (vertical) to the surfaces of the first filter sheet 12 and the second filter sheet 13, but the surrounding wall 114 is not limited to being provided in the vertical direction as long as it is provided so as to surround the microplate insertion space 115.

[0021] The portion below the bottom surface of the substrate 11 and surrounded by the surrounding wall 114 becomes a microplate insertion space 115 into which a 96-well microplate is inserted. By inserting a microplate into the microplate insertion space 115, the microplate and the substrate 11 are positioned and fixed relative to each other. The microplate insertion space 115 is basically a rectangular parallelepiped, but a corner portion 116 (see FIG. 1(c)) made of the same material as the surrounding wall 114 is provided immediately below the portion of the rectangular parallelepiped where the substrate 11 is cut off at the C-face 113. In other words, the planar shape of the microplate insertion space 115 and the planar shape of the upper surface of the substrate 11 are approximately the same.

[0022] The first filter sheet 12 is made of a filter material made of nylon 66 and having a mesh size (filtration grain size) of 30 μm. The first filter sheet 12 has approximately the same shape as the area of ​​the lower surface of the base material 11 surrounded by the surrounding wall 114, and is attached to the lower surface of the base material 11 so as to cover this area. The portion of the first filter sheet 12 facing the first through-hole 112 functions as the filter in the present invention.

[0023] The second filter sheet 13 、P A sheet material in which an adhesive sheet 132 is attached to one surface of an ET (polyethylene terephthalate) sheet 131 (one adhesive surface of a double-sided adhesive sheet is attached to a PET sheet 131) is irradiated with laser light to form a plurality of through holes (the through holes are illustrated only in FIG. 3, and are omitted in FIGS. 1 and 2). The second filter sheet 13 also has substantially the same shape as the area surrounded by the surrounding wall 114 on the lower surface of the base material 11, and is attached so as to cover the lower surface of the first filter sheet 12. The second filter sheet 13 functions as a support in the present invention. The through holes provided in the second filter sheet 13 correspond to the second through holes in the present invention. The second filter sheet 13 is formed with through holes larger than the pore size of the first filter sheet 12 and smaller than the diameter of the first through holes 112.

[0024] If the through-holes of the second filter sheet 13 are too small, the sample liquid that has passed through the first filter sheet 12 may not easily pass through the second filter sheet 13. If the through-holes of the second filter sheet 13 are too large, the area of ​​contact with the first filter sheet 12 is reduced, and the area that supports the first filter sheet 12 is narrowed. In consideration of these, the diameter of the through-holes formed in the second filter sheet 13 is preferably 0.1 μm or more and 1 mm or less. In addition, in consideration of use regardless of the pore size of the first filter sheet 12, it is more preferable that the diameter is 1 μm or more and 500 μm or less. This allows the sample liquid that has passed through the first filter sheet 12 to pass through smoothly regardless of the pore size of the first filter sheet 12, and the first filter sheet 12 to be supported more reliably.

[0025] The packing material 14 is made of a plate-like member 141 that is thicker than the first filter sheet 12 and the second filter sheet 13 and has a third through hole 142 formed in a position corresponding to the first through hole 112 of the base material 11. The packing material 14 has approximately the same planar shape as the first filter sheet 12 and the second filter sheet 13, and is provided on the lower surface of the base material 11 so as to sandwich the first filter sheet 12 and the second filter sheet 13 therebetween. The plate-like member 141 of the packing material 14 is made of silicone rubber and is flexible.

[0026] As shown in FIG. 2 and FIG. 3, the second filter sheet 13 and the packing material 14 are bonded to each other by a first double-sided adhesive film 15 having a hole at a position corresponding to the third through-hole 142 of the packing material 14 (the first double-sided adhesive film 15 is not shown in FIG. 1). In FIG. 2, the horizontal and vertical scales are different (the vertical direction is longer) in order to clearly show each component. The first filter sheet 12 and the second filter sheet 13 are bonded to each other by an adhesive sheet 132 located on the upper surface of the second filter sheet 13. The first filter sheet 12 and the base material 11 are bonded to each other by a second double-sided adhesive film 16 having a hole at a position corresponding to the first through-hole 112 of the base material 11 (the second double-sided adhesive film 16 is not shown in FIG. 1). The first double-sided adhesive film 15 and the second double-sided adhesive film 16 have the same planar shape as the first filter sheet 12, the second filter sheet 13, and the packing material 14. That is, two notches 123, 133, 143, 153, 163 are provided for each of the first filter sheet 12, the second filter sheet 13, the gasket material 14, the first double-sided adhesive film 15, and the second double-sided adhesive film 16 at locations corresponding to the two C-faces of the base material 11 (see Figure 3).

[0027] When manufacturing the filter plate 10, first, the packing material 14, the first double-sided adhesive film 15, the second filter sheet 13, the first filter sheet 12, and the second double-sided adhesive film 16 are stacked and bonded together in order from the bottom, with the notches 143, 153, 133, 123, and 163 aligned. The second filter sheet 13 is placed so that the adhesive sheet 132 faces up. Next, this integrated product (called the "filter / packing sticker 20") is fitted into the space (microplate insertion space 115) surrounded by the surrounding wall 114 on the bottom of the base material 11, with the notches aligned with the corners 116 of the base material 11, and is attached to the bottom of the base material 11 by the second double-sided adhesive film 16. This completes the filter plate 10.

[0028] A release paper may be attached to the surface of the adhesive sheet 132 (the surface on which the first filter sheet 12 is attached), and the release paper may be left attached to the adhesive sheet 132 during the manufacture of the filter plate 10, and the release paper may be peeled off from the adhesive sheet 132 during the manufacture of the seal with filter and packing material 20. Alternatively, a release paper may be attached to the back surface of the packing material 14 (the surface on which the microplate is attached), and the release paper may be left attached to the packing material 14 during the manufacture of the seal with filter and packing material 20 and the manufacture of the filter plate 10, and the release paper may be peeled off from the packing material 14 immediately before the filter plate 10 is attached to the microplate. By covering the surface of the adhesive sheet 132 and the back surface of the packing material 14 with the release paper, it is possible to prevent dust from adhering to the surface of the adhesive sheet 132 and the back surface of the packing material 14 during the manufacture of the filter plate 10, etc. Furthermore, it is possible to prevent dust from being mixed into the wells of the microplate.

[0029] Hereinafter, a method of using the filter plate 10 of the first embodiment will be described with reference to FIGS.

[0030] First, a 96-hole microplate 90 having a shape corresponding to the microplate insertion space 115 of the filter plate 10 is prepared. Then, the C surface of the microplate 90 is aligned with the corner portion 116 of the filter plate 10, and the microplate 90 is inserted from below into the microplate insertion space 115 of the filter plate 10 (FIG. 4). Next, the microplate 90 and the filter plate 10 are pressed against each other. At this time, since the packing material 14 located between the microplate 90 and the base material 11 has flexibility, the microplate 90 and the filter plate 10 are pressed against each other, and thus connected to each other in a liquid-tight state without any gaps.

[0031] Next, with the filter plate 10 fixed to the microplate 90, a predetermined amount of sample liquid 80 is dropped into each of the first through-holes 112 of the base material 11 using a micropipette (not shown). At this time, it is sufficient that the dropped sample liquid 80 is placed on the filter, and there is no need to press the tip of the tip attached to the nozzle of the micropipette strongly against the surface of the first filter sheet 12. Therefore, it is not necessary to insert the tip deeply into the first through-hole 112. For example, the task of dropping the sample liquid 80 into a plurality of first through-holes 112 simultaneously can be easily performed using a multi-channel micropipette having a plurality of nozzles.

[0032] After dropping the sample liquid 80 into the first through-hole 112 of the filter plate 10, the microplate 90 and the filter plate 10 are set in a holder 701 of the centrifuge 70 in a state where they are inclined almost vertically (see FIG. 5). In order to maintain the close contact between the microplate 90 and the filter plate 10 set in the holder 701 of the centrifuge 70, they may be fastened together with a rubber band, a clip, or the like. Thereafter, the centrifuge 70 is driven to rotate the holder 701 at high speed around the rotation axis 71 extending in the vertical direction. As a result, the sample liquid 80 in the first through-hole 112 of the filter plate 10 is pressed against the first filter sheet 12, and any matter (such as dust) larger than the pore size of the first filter sheet 12 is removed, and the sample liquid is filtered and drawn into the well 91 of the microplate 90.

[0033] There are filter plates with various pore sizes, and filter plates are used according to the purpose of use. For example, when centrifugal filtering of sample liquids derived from living organisms or capturing intracellular substances (nucleic acids, etc.), filters with small pore sizes and thin filters are often used. Since filter plates with small pore sizes are thin and weak, conventional filter plates can break when centrifugal filtering is performed.

[0034] In contrast, in the filter plate 10 of this embodiment, the second filter sheet 13 is disposed adjacent to the lower surface (the side from which the sample liquid flows out) of the first filter sheet 12 used for the purpose of filtering the sample. The second filter sheet 13 is not mesh-like, but is a sheet material having a plurality of through holes formed therein, and is in contact with the first filter sheet 12 by a surface, not by a point or a line. Therefore, when centrifugal filtration is performed, the first filter sheet 12 is supported by the second filter sheet 13 from the back side (the side of the microplate 90), and damage to the first filter sheet 12 is suppressed. Furthermore, the second filter sheet 13 has through holes formed therein that are larger than the pore size of the first filter sheet, so that the sample liquid that has passed through the first filter sheet 12 is not prevented from being drawn into the wells 91 of the microplate 90.

[0035] Furthermore, since the filter plate 10 and the microplate 90 are connected in a liquid-tight state by the flexible packing material 14, when the sample liquid 80 in the first through-hole 112 is drawn into the wells 91 by centrifugal force, the sample liquid is prevented from flowing into other wells 91. Therefore, it is possible to prevent different sample liquids from being supplied to the wells 91 in a mixed state.

[0036] (2) Second embodiment Fig. 6 and Fig. 7 show a filter plate 30 of the second embodiment. Fig. 6(a) is a top view of the filter plate 30, Fig. 6(b) is a vertical cross-sectional view of the filter plate 30 taken along line bb, and Fig. 6(c) is a bottom view of the filter plate 30. Fig. 7(a) is a partially enlarged view of Fig. 6(b), and Fig. 7(b) is a partially enlarged view of Fig. 6(c).

[0037] The filter plate 30 includes a base material 31 , a filter sheet 32 ​​, a reinforcing sheet material 33 , a packing material 34 , and a cylindrical body 37 .

[0038] The base material 31 is made of a plastic plate-like member having a rectangular planar shape, and has 24 first through-holes 312 arranged parallel to the long sides and 16 first through-holes 312 arranged parallel to the short sides, for a total of 384 first through-holes 312 having a circular planar shape. The arrangement of the first through-holes 312 corresponds to the arrangement of the wells of a 384-well microplate to which the filter plate 30 is attached.

[0039] The substrate 31 is composed of a first substrate 3111 and a second substrate 3112 attached to the lower part of the first substrate 3111. A rectangular frame-shaped surrounding wall 3114 extends downward from the outer periphery of the lower surface of the first substrate 3111. In this embodiment as well, the surrounding wall 3114 is provided perpendicular (vertical) to the surfaces of the filter sheet 32 ​​and the reinforcing sheet material 33, but the surrounding wall 3114 is not limited to being vertical as long as it is provided so as to surround the microplate insertion space 115.

[0040] The filter sheet 32, the reinforcing sheet material 33, and the second base material 3112 are attached in this order from the first base material 3111 side to a portion surrounded by the surrounding wall 3114 on the lower surface of the first base material 3111. The length of the surrounding wall 3114 is greater than the combined thickness of the filter sheet 32, the reinforcing sheet material 33, and the second base material 3112, and the space below the second base material 3112 and surrounded by the surrounding wall 3114 becomes the microplate insertion space 315. The first base material 3111 and the second base material 3112 each have 384 through holes 3121, 3122, and the first through holes 312 are constituted by the through holes 3121, 3122 that are positioned correspondingly when the first base material 3111 and the second base material 3112 are stacked vertically. Through-hole 3121 is cylindrical with a uniform inner diameter in the depth direction (the vertical direction in FIG. 7(a)), whereas through-hole 3122 has a tapered shape with an inner diameter smaller at the bottom than at the top.

[0041] A cylinder 37 is provided on the lower surface of the second base material 3112 so as to extend downward from the periphery of the first through-hole 312. The cylinder 37 is made of the same plastic as the second base material 3112 and is molded integrally with the second base material 3112. The outer diameter of the cylinder 37 is slightly smaller than the inner diameter of the wells of the 384-well microplate, and the inner diameter of the cylinder 37 is equal to the inner diameter of the through-hole 3122 on the lower surface of the second base material 3112. The length of the cylinder 37 is shorter than the depth of the wells of the 384-well microplate.

[0042] The filter sheet 32 ​​and the reinforcing sheet material 33 are interposed between the first base material 3111 and the second base material 3112. The material and the size of the openings of the filter sheet 32 ​​are the same as those of the first filter sheet 12 of the first embodiment. The reinforcing sheet material 33 (corresponding to the support in the present invention) is, for example, a sheet material in which an adhesive sheet is attached to the surface of a sheet made of a resin harder than the silicone rubber constituting the packing material 34 described below, and similarly to the second filter sheet 13 in the first embodiment, a large number of through holes (corresponding to the second through holes in the present invention) having a diameter larger than the pore size of the filter sheet 32 ​​and smaller than the diameter of the first through holes 312 are formed.

[0043] Since the reinforcing sheet material 33 has a through hole with a diameter larger than the pore size of the filter sheet 32, the reinforcing sheet material 33 does not hinder the flow of the sample liquid that has passed through the filter sheet 32. Furthermore, since the reinforcing sheet material 33 has a through hole with a diameter smaller than the diameter of the first through hole 312, the filter sheet 32 ​​can be supported inside the first through hole 312 regardless of the position where the through hole is formed. In the second embodiment, for the reasons described in the first embodiment, the diameter of the through hole is preferably 0.1 μm or more and 1 mm or less, and more preferably 1 μm or more and 500 μm or less. As in the first embodiment, the filter sheet 32 ​​and the reinforcing sheet material 33 cover the entire upper surface of the second base material 3112, and the part of the filter sheet 32 ​​located in the first through hole 312 functions as a filter in the present invention.

[0044] The packing material 34 is made of a plate-like member made of silicone rubber, and is attached to the second base material 3112 so as to cover the lower surface of the second base material 3112 except for the cylindrical body 37. Therefore, the packing material 34 has a third through hole 342 corresponding to the cylindrical body 37.

[0045] Although detailed explanation and illustration are omitted, the first base material 3111 and the filter sheet 32, the reinforcing sheet material 33 and the second base material 3112, and the second base material 3112 and the packing material 34 are bonded together with an adhesive or an adhesive sheet with adhesive applied to both sides. The filter sheet 32 ​​and the reinforcing sheet material 33 are bonded together with an adhesive sheet that the reinforcing sheet material 33 has.

[0046] The filter plate 30 is used as follows. First, a microplate 90A (384 holes) having 384 wells 91A is prepared. Then, as shown in FIG. 8, the microplate 90A is inserted from below into the microplate insertion space 315 of the filter plate 30. At this time, the cylinder 37 corresponding to the position is inserted into each well 91A of the microplate 90A. Next, the microplate 90A and the filter plate 30 are pressed against each other. By pressing the microplate 90A and the filter plate 30 against each other in this manner, the flexible packing material 34 is tightly adhered to the upper surface of the microplate 90A without any gaps, and the filter plate 30 and the microplate 90A are connected in a liquid-tight state. The operations from dropping the sample liquid 80 into the first through-hole 312 to supplying the sample liquid 80 to the wells 91A by applying centrifugal force are the same as those in the first embodiment.

[0047] In the second embodiment, similarly to the first embodiment, a reinforcing sheet material 33 is disposed adjacent to the lower surface (the side from which the sample liquid flows out) of the filter sheet 32. Moreover, the reinforcing sheet material 33 is not mesh-shaped, but is a sheet material having a plurality of through holes formed therein, and is in contact with the filter sheet 32 ​​by a surface, not by a point or a line. Therefore, when centrifugal filtration is performed, the filter sheet 32 ​​is supported from the back side (the side of the microplate 90A) by the reinforcing sheet material 33, and damage to the filter sheet 32 ​​is suppressed. Moreover, since the reinforcing sheet material 33 has through holes larger than the pore size of the first filter sheet, the sample liquid that has passed through the filter sheet 32 ​​is not prevented from being introduced into the well 91A of the microplate 90A.

[0048] Furthermore, since the filter plate 30 is connected in a liquid-tight state to the microplate 90A by the flexible packing material 34, when the sample liquid 80 dropped into the first through-hole 312 is drawn into the microplate 90A by centrifugal force, the sample liquid is prevented from leaking out and flowing into other wells 91A. Furthermore, since the cylinder 37 is provided on the lower surface of the base material 31 of the filter plate 30, the liquid sample that has passed through the filter sheet 32 ​​can be guided to the vicinity of the bottom of the well 91A of the microplate 90A.

[0049] In the filter plate 30 of the second embodiment, the filter sheet 32 ​​is sandwiched and fixed between the first base material 3111 and the second base material 3112 together with the reinforcing sheet material 33, and the filter is tightly stretched in each of the first through holes 312. Thus, in addition to filtering sample liquid, the filter plate 30 can be used to collect intracellular substances such as intracellular nucleic acids and exosomes contained in blood or cell culture fluid.

[0050] When recovering intracellular substances from cells contained in a sample liquid, a filter sheet 32 ​​with a pore size of 1 μm or less is used, and the holder 701 of the centrifuge 70 is rotated at a higher speed than when filtering the sample liquid. This generates a large centrifugal force, and the cells in the sample liquid are strongly pressed against the filter sheet 32. Since the filter sheet 32 ​​is in a state in which the filter sheet 32 ​​is strongly stretched on each of the first through holes 312 of the filter plate 30, a large normal force acts on the cells strongly pressed against the filter sheet 32, and the cells can be destroyed by the centrifugal force and the normal force. The destroyed cells and intracellular substances can be separated by appropriately setting the mesh size of the filter sheet 32. Alternatively, nucleic acids in the cells can be captured by using an appropriate filter sheet 32. In this way, when an operation is performed in which the filter sheet 32 ​​with a pore size of 1 μm or less is strongly stretched and the cells are strongly pressed against the filter sheet 32, the filter sheet 32 ​​is particularly likely to be damaged. However, in the second embodiment, since the filter sheet 32 ​​is supported by the reinforcing sheet material 33, damage to the filter can be prevented.

[0051] (3) Modifications The present invention is not limited to the above-described embodiment, and various modifications are possible.

[0052] The materials of the base material 11, 31, the first filter sheet 12, 32, the second filter sheet 13, the reinforcing sheet material 33, and the packing material 14, 34 shown in each of the above embodiments are merely examples, and other materials may be used. For example, the packing material 14, 34 may be made of ethylene propylene rubber (EPM (EPR) or EPDM (EPT)), urethane rubber, etc., instead of silicone rubber. Furthermore, the first filter sheet 12, 32 may be made of a polymer material such as polyester, polyethylene, polypropylene, etc., instead of nylon 66, or may be made of a material other than a polymer material, such as metal. The PET sheet 131 of the second filter sheet 13 may be replaced by a sheet made of another material.

[0053] The opening of the first filter sheet 12, 32 can be appropriately set according to the intended use of the filter plate 10, 30. For example, the opening can be 1 μm, 10 μm, 40 μm, 70 μm, 100 μm, etc. Alternatively, a precision filtration filter (membrane filter) with an opening of less than 1 μm (so-called submicron) may be used. For example, when exosomes are collected as intracellular substances, the opening of the filter sheet may be set to 0.1 μm to 5 μm. In either case, the second filter sheet 13 may have through holes formed therein that are larger than the pore size of the first filter sheet 12, 32 and smaller than the diameter of the first through hole 112, and the reinforcing sheet material 33 may have through holes formed therein that are larger than the pore size of the first filter sheet 12, 32 and smaller than the diameter of the first through hole 312.

[0054] As a component corresponding to the support in the present invention, the second filter sheet 13 is used in the first embodiment, and the reinforcing sheet material 33 is used in the second embodiment, but various forms of supports can be used. However, it is preferable to use a support that contacts the first filter sheets 12, 32 with a surface, not a point or line (i.e., supports the first filter sheets 12, 32 with a surface). In addition, the second filter sheet 13 and the packing material 14 in the first embodiment, and the reinforcing sheet material 33 and the packing material 34 in the second embodiment may be configured as a single member. In that case, the support is configured with a material having flexibility to function as a packing material. In addition, since it is expected that the through-holes will deform when the filter plate is pressed against the microplate, in order to prevent the flow of the sample liquid from being hindered by this, it is preferable to make the size of the through-holes formed in the support larger than those described in the first and second embodiments.

[0055] In the first and second embodiments, the second filter sheet 13 and the reinforcing sheet material 33 are made of adhesive sheets that have been previously attached, but the PET sheet 131 and the adhesive sheet may be prepared separately. In the first and second embodiments, the seal with the filter and packing material is integrally constructed using a double-sided adhesive film or an adhesive sheet, but these do not necessarily have to be integrated. As in the above embodiments, each member has a notch, so that if the members are inserted into the microplate insertion space 115, 315 in order while aligning the positions of the notches, each member can be aligned without integrally constructing the seal with the filter and packing material using a double-sided adhesive film.

[0056] In the first embodiment, a filter plate having first and third through holes whose number and positions correspond to wells 91 of a 96-well microplate 90 is shown, and in the second embodiment, a filter plate having first and third through holes whose number and positions correspond to wells 91A of a 384-well microplate 90A is shown, but the filter plate 10 of the first embodiment and the filter plate 30 of the second embodiment can be applied to various microplates by providing first and third through holes whose number and positions correspond to wells 91, 91A of the microplates 90, 90A. Furthermore, the present invention is not limited to microplates, but can also be applied to dishes, tubes, bottles, flasks, bags, etc. is applicable to.

[0057] In the first and second embodiments, the first through hole (substrate) and the third through hole (gasket material) are provided at positions corresponding to all of the wells 91, 91A of the microplate 90, 90A, and the filter sheets 12, 13, 32 and the reinforcing sheet material 33 are arranged to cover all of those wells (sample storage sections). However, when only some of the wells (a portion of the multiple sample storage sections) are used, the first through hole (substrate) and the third through hole (gasket material) may be provided only at positions corresponding to those portions of the wells, and the filter sheets 12, 13, 32 and the reinforcing sheet material 33 may be arranged to cover only the positions corresponding to those portions of the wells.

[0058] In the filter plate of the first embodiment, the corner portion 116 is provided in the microplate insertion space 115 of the base material 11, but the corner portion 116 may be omitted. By omitting the corner portion 116, the filter plate can be used for a microplate that does not have a C-face.

[0059] In the first and second embodiments, the filters corresponding to all the first through holes are constructed from a single first filter sheet 12, 32, and a single second filter sheet 13 and a reinforcing sheet material 33 are arranged on the back surface of the single first filter sheet 12, 32. However, as in the filter plate 10A shown in FIG. 9, each of the multiple first through holes 112 may be provided with an individual first filter 12A and second filter 13A (a filter having a pore size larger than the pore size of the first filter 12A).

[0060] In the above embodiment, the planar shape of the first through holes 112, 312 is circular, but it may be a quadrangle such as a square, a hexagon such as a regular hexagon, or other shapes. The diameter of the through hole described in the first and second embodiments is for a case where the through hole is circular, and in the case of a through hole other than a circle, the through hole may have a size and shape that has the same area as the area of ​​a circle with the above diameter. In addition, the shape of the vertical cross section of the first through holes 112, 312 is not limited to the above example.

[0061] (4) Related embodiments Next, a filter plate 40 according to a related embodiment will be described. The same components as those in the first embodiment are denoted by the same reference numerals, and descriptions thereof will be omitted as appropriate.

[0062] Fig. 10 shows a longitudinal cross-sectional view of a filter plate 40 of a related embodiment. Fig. 10 corresponds to the longitudinal cross-sectional view taken along line aa in Fig. 1(a) described in the first embodiment. This filter plate 40 has a base material 11, a first filter sheet 42, a second filter sheet 43, and a packing material 14. The base material 11 and the packing material 14 are the same as those in the first embodiment.

[0063] Fig. 11 is a vertical cross-sectional view showing a filter / packing seal 50 including a filter and packing, which is attached to a base material during the manufacture of the filter plate 40 of the related embodiment, and Fig. 12 is an exploded perspective view of the filter / packing seal 50.

[0064] The first filter sheet 42 is the same member as the first filter sheet 12 in the first embodiment (however, the arrangement is different from that in the first embodiment). The second filter sheet 43 is a sheet material (one adhesive surface of a double-sided adhesive sheet is attached to the PET sheet 431) in which an adhesive sheet 432 is attached to one surface of a PET (polyethylene terephthalate) sheet 431) and a plurality of through holes are formed by irradiating the sheet material with laser light. However, unlike the first embodiment, as shown in the partially enlarged vertical cross-sectional view of the second filter sheet 43 in FIG. 13, the adhesive sheet 432 is located below the PET sheet 431. In addition, a tapered through hole 433 is formed, the diameter of which decreases from the upper side to the lower side (the adhesive sheet 432 side).

[0065] As shown in FIG. 11 and FIG. 12, the first filter sheet 42 and the packing material 14 are bonded to each other by a first double-sided adhesive film 45 having a hole at a position corresponding to the third through hole 142 of the packing material 14 (the first double-sided adhesive film 45 is not shown in FIG. 10). The first double-sided adhesive film 45 may be the same as the first double-sided adhesive film 15 in the first embodiment. The first filter sheet 42 and the second filter sheet 43 are bonded to each other by an adhesive sheet 432 located on the lower surface of the second filter sheet 43. The second filter sheet 43 and the base material 11 are bonded to each other by a second double-sided adhesive film 46. The second double-sided adhesive film 46 may be the same as the first double-sided adhesive film 16 in the first embodiment. The first double-sided adhesive film 45 and the second double-sided adhesive film 46 have the same planar shape as the first filter sheet 42, the second filter sheet 43, and the packing material 14. That is, two notches 423, 433, 143, 453, 463 are provided for each component at locations of the first filter sheet 42, the second filter sheet 43, the gasket material 14, the first double-sided adhesive film 45, and the second double-sided adhesive film 46 corresponding to the two C-faces of the base material 11 (see Figure 3).

[0066] A method of using the filter plate 40 of the related embodiment will now be described.

[0067] First, a 96-hole microplate is prepared in a shape corresponding to the microplate insertion space 115 of the filter plate 40. Then, the C surface of this microplate is aligned with the corner portion 116 of the filter plate 40, and the microplate is inserted from below into the microplate insertion space 115 of the filter plate 40. Next, the microplate and the filter plate 40 are pressed against each other. At this time, since the packing material 14 located between the microplate and the base material 11 has flexibility, the microplate 90 and the filter plate 40 are pressed against each other, and thus the two are connected in a liquid-tight state without any gaps.

[0068] Next, a pipette tip 60 is attached to each of the nozzles of the multichannel micropipette, and the sample liquid is aspirated therein. Then, each pipette tip 60 is inserted into the first through-hole 112 of the base material 11, and the tip of each pipette tip 60 is pressed against the second filter sheet 43 to discharge the sample liquid 80. This allows the sample liquid 80 to be dispensed simultaneously into the first through-holes 112 corresponding to the wells 91 arranged in a row. FIG. 14 is a partially enlarged view showing how the pipette tip 60 is inserted into one of the first through-holes 112 and pressed against the second filter sheet 43 to extract the sample liquid 80. This causes the sample liquid 80 to be pushed out toward the first filter sheet 42, and the sample liquid 80 can be filtered by the first filter sheet 42 in a short time without using a centrifuge 70. Here, a preferred example of efficiently dispensing the sample liquid 80 using a multichannel pipette having multiple nozzles has been described, but a pipette having only one nozzle may be used.

[0069] As described above, when the tip of the tip is pressed directly against the thin filter sheet to dispense the sample liquid 80, the filter sheet is easily damaged. In contrast, in the related embodiment, the second filter sheet 43 is disposed on the upper part of the first filter sheet 42, and the tip of the pipette tip 60 is configured not to directly touch the first filter sheet 42, so that the first filter sheet 42 can be prevented from being damaged. In addition, the through holes provided in the second filter sheet 43 are tapered so that the diameter becomes smaller from the upper part (the side where the sample liquid 80 flows in) to the lower part (the side where the sample liquid 80 flows out). That is, the flow path of the sample liquid 80 gradually narrows. Therefore, the sample liquid 80 discharged from the tip of the pipette tip 60 is pushed against the first filter sheet 42 with a strong force, and the sample liquid can be filtered without using a centrifuge. The thickness of the second filter sheet 43 may be appropriately set according to the strength of the first filter sheet 42, the diameter of the through holes formed in the second filter sheet 43, and the like.

[0070] [Aspects] It will be appreciated by those skilled in the art that the exemplary embodiments described above are examples of the following aspects.

[0071] (Section 1) One aspect of the present invention is a filter plate that is attached to a sample container having a sample storage portion with an open top, comprising: a) a substrate having a first through hole through which a sample liquid flows, the first through hole being provided at a position corresponding to the sample storage portion; b) a first filter provided in the first through hole; c) a support disposed adjacent to the first filter on the side from which the sample liquid flows out, the support having second through-holes formed therein and having a pore size larger than that of the first filter; Equipped with.

[0072] The filter plate according to the first aspect is fixed to a sample container so that the position of the opening of the sample container and the position of the first through-hole of the base coincide with each other, and the support is located closer to the sample container than the filter. In the filter plate according to the present invention, a filter used for centrifugal filtration is provided in the first through-hole of the base used by fixing to the sample container. The filter plate according to the first aspect further includes a support disposed adjacent to the side of the first filter from which the sample liquid flows out. Therefore, when performing an operation in which force is applied to the filter, such as centrifugal filtration, the filter is supported from the back side (the side of the sample container), and damage to the first filter is suppressed. In addition, the support is formed with a second through-hole larger than the pore size of the first filter used for centrifugal filtration, and thus the sample liquid that has passed through the filter is not prevented from being introduced into the wells of the microplate.

[0073] (Section 2) The filter plate according to paragraph 2 is the filter plate according to paragraph 1, The filter plate is used for a microplate having a plurality of wells, and the first through-holes are provided in a base material at positions corresponding to each well of the microplate; The first filter is a sheet-like filter disposed on one surface of the base so as to cover all of the first through-holes.

[0074] The filter plate according to paragraph 1 can be used in a microplate having multiple wells, as described in paragraph 2. In that case, as described in paragraph 2, the first filter can be easily attached by using a sheet-like filter arranged on one surface of the base so as to cover all of the first through-holes.

[0075] (Section 3) The filter plate according to paragraph 3 is a filter plate according to paragraph 1 or 2, The second through hole has a smaller diameter than the first through hole.

[0076] In the filter plate according to the third aspect, the filter can be supported by the support body inside the first through hole, regardless of the position at which the second through hole is provided in the support body.

[0077] (Section 4) The filter plate according to paragraph 4 is a filter plate according to any one of paragraphs 1 to 3, The support is a second filter formed by bonding an adhesive sheet to a sheet-like material and forming the second through holes therein.

[0078] In the filter plate according to the fourth aspect, the first filter and the second filter are adhered to each other by the adhesive sheet of the support, so that the sample liquid can be prevented from flowing between the two filters and causing leakage.

[0079] (Section 5) The filter plate according to paragraph 5 is a filter plate according to any one of paragraphs 1 to 4, The second through hole has a diameter of not less than 0.1 μm and not more than 1 mm.

[0080] If the second through hole is too small, the sample liquid that has passed through the first filter may not easily pass through the second filter. If the second through hole is too large, the contact area between the first filter and the second filter is reduced, and the area that can support the first filter is narrowed. In consideration of these, it is preferable that the diameter of the second through hole is set to the range specified in item 6. Note that this is on the assumption that the through hole has a circular cross section, and in the case of a through hole other than a circular cross section, the through hole may be of a size and shape that has the same area as the area of ​​a circle with the above diameter.

[0081] (Section 6) The filter plate according to paragraph 6 is the filter plate according to any one of paragraphs 1 to 5, further comprising: a flexible, plate-like packing material that is disposed on the side of the support from which the sample liquid flows out and has third through-holes formed at positions corresponding to each of the plurality of wells; Equipped with.

[0082] In the filter plate according to the sixth aspect, the sample liquid flowing in from the first through-hole of the substrate can be introduced into the well corresponding to the first through-hole without leakage. [Explanation of symbols]

[0083] 10, 10A, 30, 40… Filter plate 11...Base material 111...Plate-shaped member 112...First through hole 113…C side 114…Enclosure wall 115...Microplate insertion space 116…Corner section 12, 42…First filter sheet 12A…First filter 13, 43…Second filter sheet 131, 431...PET sheet 132, 432...adhesive sheet 13A…Second filter 14…Packing material 141...Plate-shaped member 142…Third through hole 15…First double-sided adhesive film 16…Second double-sided adhesive film 20, 50... Filter and packing seal 31...Base material 3111…First base material 3112…Second base material 3114…Enclosure wall 312…First through hole 3121: Through hole of first substrate 3122: Through hole of second substrate 315...Microplate insertion space 32...Filter sheet 33…Reinforcing sheet material 34…Packing material 342…Third through hole 37...Cylinder 433... through hole of second filter sheet (second through hole) 60…Pipette tip 70...Centrifuge 701...Holder 71...Rotation axis 80...Sample liquid 90, 90A...Microplate 91, 91A…Well

Claims

1. A filter plate to be attached to a sample container having a sample receiving portion with an open top, a) a substrate having a first through-hole for passing a sample liquid at a position corresponding to the sample storage portion; b) a first filter provided in the first through hole; c) a second filter arranged adjacent to the first filter on the side from which the sample liquid flows out, the second filter being formed by laminating an adhesive sheet to a sheet-like material and having second through-holes larger than the pore size of the first filter, the first filter being adhered to the adhesive surface of the adhesive sheet; A filter plate comprising:

2. the filter plate is used in a microplate having a plurality of wells, and the first through-holes are provided in the base material at positions corresponding to the respective wells of the microplate; The first filter is a sheet-like filter disposed on one surface of the substrate so as to cover all of the first through holes.

2. The filter plate according to claim 1, characterized in that:

3. The diameter of the second through hole is smaller than the diameter of the first through hole.

2. The filter plate according to claim 1, characterized in that:

4. The diameter of the second through hole is 0.1 μm or more and 1 mm or less.

2. The filter plate according to claim 1, characterized in that:

5. a flexible, plate-shaped packing material disposed on the side of the second filter from which the sample liquid flows out, the packing material having third through-holes formed at positions corresponding to the plurality of wells; 2. The filter plate of claim 1, comprising:

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