WO2014109323A1 - 液体充填補助具および液体充填方法 - Google Patents
液体充填補助具および液体充填方法 Download PDFInfo
- Publication number
- WO2014109323A1 WO2014109323A1 PCT/JP2014/050100 JP2014050100W WO2014109323A1 WO 2014109323 A1 WO2014109323 A1 WO 2014109323A1 JP 2014050100 W JP2014050100 W JP 2014050100W WO 2014109323 A1 WO2014109323 A1 WO 2014109323A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- liquid
- liquid filling
- reaction
- filling aid
- main body
- Prior art date
Links
- 239000007788 liquid Substances 0.000 title claims abstract description 212
- 238000000034 method Methods 0.000 title claims abstract description 25
- 238000004891 communication Methods 0.000 claims abstract description 41
- 238000007599 discharging Methods 0.000 claims abstract description 3
- 239000011521 glass Substances 0.000 claims description 56
- 238000010186 staining Methods 0.000 claims description 16
- 230000009471 action Effects 0.000 claims description 12
- 238000004043 dyeing Methods 0.000 claims description 12
- 239000007769 metal material Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 abstract description 5
- 238000001704 evaporation Methods 0.000 abstract description 5
- 230000008020 evaporation Effects 0.000 abstract description 5
- 239000012530 fluid Substances 0.000 abstract description 4
- 238000010438 heat treatment Methods 0.000 description 13
- 230000008569 process Effects 0.000 description 12
- MOVRNJGDXREIBM-UHFFFAOYSA-N aid-1 Chemical compound O=C1NC(=O)C(C)=CN1C1OC(COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C(NC(=O)C(C)=C2)=O)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C(NC(=O)C(C)=C2)=O)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C(NC(=O)C(C)=C2)=O)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)COP(O)(=O)OC2C(OC(C2)N2C3=C(C(NC(N)=N3)=O)N=C2)CO)C(O)C1 MOVRNJGDXREIBM-UHFFFAOYSA-N 0.000 description 10
- 238000004140 cleaning Methods 0.000 description 10
- 238000011532 immunohistochemical staining Methods 0.000 description 7
- 239000000427 antigen Substances 0.000 description 6
- 108091007433 antigens Proteins 0.000 description 6
- 102000036639 antigens Human genes 0.000 description 6
- 239000013076 target substance Substances 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 230000004913 activation Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 230000000630 rising effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 238000009412 basement excavation Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000000428 dust Substances 0.000 description 2
- 239000011536 extraction buffer Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 1
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000003028 elevating effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 108700020302 erbB-2 Genes Proteins 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 239000004570 mortar (masonry) Substances 0.000 description 1
- 238000012758 nuclear staining Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N1/31—Apparatus therefor
- G01N1/312—Apparatus therefor for samples mounted on planar substrates
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/52—Containers specially adapted for storing or dispensing a reagent
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L9/00—Supporting devices; Holding devices
- B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L9/00—Supporting devices; Holding devices
- B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
- B01L9/527—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips for microfluidic devices, e.g. used for lab-on-a-chip
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
- G01N35/1002—Reagent dispensers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/025—Align devices or objects to ensure defined positions relative to each other
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0621—Control of the sequence of chambers filled or emptied
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0684—Venting, avoiding backpressure, avoid gas bubbles
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/046—Function or devices integrated in the closure
- B01L2300/047—Additional chamber, reservoir
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/04—Closures and closing means
- B01L2300/046—Function or devices integrated in the closure
- B01L2300/048—Function or devices integrated in the closure enabling gas exchange, e.g. vents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
- B01L2300/0822—Slides
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0877—Flow chambers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
Definitions
- the present invention relates to a liquid filling aid and a liquid filling method, and more specifically, for example, a liquid filling aid used in a process of staining biological tissue or cells adhered or pasted on a slide glass, and the liquid filling
- the present invention relates to a liquid filling method using an auxiliary tool.
- the immunohistochemical staining method is a method for clarifying the localization (location) of a substance having antigenicity in a tissue / cell using a specific immune reaction called an antigen-antibody reaction.
- the operation of immunohistochemical staining generally comprises a plurality of steps as follows.
- the first step is a deparaffinization step in which paraffin embedding a specimen fixed using a formalin solution or the like affixed on a slide glass is dissolved in an organic solvent
- the second step is to remove the organic solvent
- third step is antigen activation step for performing treatment for enhancing immunoreactivity
- fourth step is a primary antibody to which an antibody that reacts with an antigen in a sample is applied
- the application step is a detection step for applying a reagent for detecting the reacted antibody
- the sixth step is a color development step for applying a reagent for visualizing the detection reagent
- the seventh step is: In order to make the shape of the tissue easy to see, this is a nuclear staining process in which the nucleus is stained in different colors.
- Patent Document 1 discloses a main body part that is located above a base material and defines a cavity for forming a reaction chamber, and extends from the main body part when the cover is attached to the base material. And a cover for the substrate including a protrusion defining a fluid container in fluid communication with the cavity.
- Patent Document 2 discloses means for adding a processing liquid to the surface of a slide carrying a sample, and means for determining a contour of a sample by installing a cover tile having a concave surface so as to cover the sample on the slide.
- An apparatus for treating a human or animal cell sample with a treatment liquid wherein the apparatus further seals the sealing liquid between the slide and the cover tile so as to contain a gap formed between the cover tile and the slide.
- a device including means for adding to the joint from outside the cavity.
- Patent Document 3 includes a base having an upper surface and a bottom surface, a base holding a tissue specimen on the upper surface of the base, and a slide cover installed on the upper surface of the base, having an inner surface
- An apparatus for extracting biomolecules from a tissue specimen including a slide cover in which a top surface of the slide cover and an inner surface of the slide cover form a space for holding an extraction buffer, and a temperature control device connected to the bottom surface of the base is disclosed.
- the inner surface of the slide cover protrudes toward the base at the center of the slide cover, so that the space formed between the upper surface of the base and the inner surface of the slide cover is shallow around the center.
- the slide cover In order to add the extraction buffer to the space between the upper surface of the base and the inner surface of the slide cover, the slide cover has an opening at the center (Claim 15), Cover has to contain at least one lateral holes (Claim 16), it is disclosed further around part.
- Patent Document 1 and Patent Document 2 have the following problems.
- an additional mechanism for assisting filling is required, such as sliding the cover or sucking from the side opposite to the discharge side.
- the whole space enclosed by the cover and the base part cannot be filled with the liquid by the capillary action in the first place.
- an object of the present invention is to provide a liquid filling aid and a liquid filling method that solve the above-described problems.
- 1st invention is a liquid filling auxiliary
- the liquid filling aid is characterized by comprising a vent hole (14), and the reaction section and the upper surface of the plate-like body constitute a reaction chamber (17).
- the second invention is characterized in that, in the first invention, the volume of the storage part (2) is larger than the volume of the reaction part (8).
- the storage part (2) is arranged above the reaction part (8) with the communication hole interposed therebetween.
- the fourth invention is characterized in that, in the third invention, an opening (3) of the reservoir is provided on the upper surface of the main body (30).
- the communication hole (15) may be located substantially at the center of the reaction part (8).
- the vent hole (14) penetrates from the upper surface of the reaction section (8) to the upper surface of the main body (30).
- a sixth invention is characterized in that, in the first invention, the reaction section (8) is configured to a depth at which a force due to capillary action is generated in the entire reaction section. According to a seventh aspect, in the sixth aspect, the depth is 10 to 200 ⁇ m.
- the eighth invention is characterized in that, in the first invention, the bottom surface of the storage portion (2) has an inclined surface or a groove toward the communication hole.
- at least one of the inclined surfaces facing the communication hole (15) has two or more inclinations.
- the storage portion (2) includes a first recess (24) in which the communication hole is provided substantially at the center, and is positioned above the first recess and has a volume. And a second recess that is twice or more in size.
- an inner upper surface (ceiling surface) of the reaction part (8) has an inclined surface or a groove toward the vent hole, and the vent hole (14) is provided in the reaction part.
- a twelfth invention is characterized in that, in any one of the first to eleventh inventions, the main body (30) is made of a metal material.
- a thirteenth invention is characterized in that, in any one of the first to eleventh inventions, the plate-like body is a slide glass used in a staining step.
- a fourteenth aspect of the invention is a liquid discharge device, a pallet that holds a slide glass, a drive device that allows the discharge device to move relative to the pallet in the XYZ directions, and a liquid filling aid according to the eleventh aspect of the invention.
- It is an automatic dyeing
- a stage having a heating function and an elastic body that urges the pallet upward may be provided, and the pallet may include an opening through which the stage can pass when the elastic body contracts. .
- a fifteenth aspect of the invention is a liquid filling method using the liquid filling aid according to any one of the first to eleventh inventions, the step of installing the liquid filling aid on a plate-like body, and the reaction chamber And a step of discharging a liquid having a volume larger than the volume to the reservoir.
- the liquid can be filled to every corner of the reaction part only by the force due to the capillary phenomenon. Further, since the liquid stored in the storage part flows down from the communication hole to the reaction part by its own weight, an additional mechanism for assisting the filling is not necessary.
- this liquid filling auxiliary tool is installed on the slide glass, the reaction part constitutes a reaction chamber which is a closed space, and thus drying can be prevented. Moreover, by filling the reaction part with the liquid until it reaches the vent hole, the contact area between the liquid and the air can be reduced and the drying can be delayed.
- FIG. 2 is a cross-sectional view taken along line AA in FIG. It is a bottom view of the liquid filling auxiliary tool of the first embodiment.
- (a) is a state immediately after the liquid is discharged to the storage part
- (b) is a state in which the liquid flows from the storage part through the communication hole to the reaction part
- (c) is a liquid filling the entire reaction part.
- Each state is shown.
- FIG. 1 is a top view of the liquid filling aid 1 of the present embodiment
- FIG. 2 is a cross-sectional view taken along line AA in FIG. 1
- FIG. 3 is a bottom view of the liquid filling aid 1 of the present embodiment.
- the horizontal direction in FIG. 1 or 3 may be referred to as the width direction
- the vertical direction as the depth direction
- the vertical direction in FIG. 2 as the height direction.
- the liquid filling auxiliary tool 1 of the present embodiment is configured by forming a storage part 2, a reaction part 8, a vent hole 14, and a communication hole 15 in a main body 30 which is a rectangular parallelepiped member, and will be described later. It is installed on the glass 16 and used.
- the length of the liquid filling auxiliary tool 1 in the depth direction is the same as or slightly shorter than the length of the short side of the slide glass 16.
- the length in the width direction is about 2/3 of the length of the long side of the slide glass 16. This length is appropriately changed corresponding to the usable area of the slide glass 16.
- the remaining about one third of the slide glass 16 that is not covered with the liquid filling auxiliary tool 1 is a space where characters, symbols, bar codes, and the like for identification are written.
- the shape of the main body 30 is not limited to the illustrated rectangular parallelepiped shape, and may be a polygonal column, a cylinder, or an elliptical column. Hereinafter, each configuration will be described in detail.
- the reservoir 2 is recessed so that the upper surface opens on the upper surface side of the main body 30 and has a capacity several times to several tens of times the capacity of the reaction chamber 17 (for example, a capacity of about 60 times).
- the illustrated storage unit 2 has a rectangular shape when viewed from the upper surface, but is not limited thereto, and the shape viewed from the upper surface may be a polygon, a circle, or an ellipse.
- the storage unit 2 preferably has a depth of 1 ⁇ 2 or more of the height of the main body 30 (cuboid), more preferably 2/3 or more. Further, the width and depth (area of the opening 3) are set to be slightly smaller than the area of the upper surface of the rectangular parallelepiped, leaving the wall 4 forming the storage portion 2.
- the width and depth of the reaction portion 8 are made smaller by the location where the vent hole 14 is provided.
- staining reaction is not limited to these, It can change suitably. However, it is preferably larger than the capacity of the reaction section 8. This is for the purpose of accumulating the amount required for the reaction unit 8 and the additional required amount at a time.
- a liquid for activating the antigen, a solution containing an antibody that reacts with the antigen, distilled water, an organic solvent, or the like is discharged from the discharge device 18 to the storage unit 2.
- the inner and lower surfaces of the reservoir 2 are constituted by two inclined surfaces 5 and 5 that descend from the short sides 7 and 7 toward the center. And one communication hole 15 is provided in the center of the lowest part 6 where the two inclined surfaces 5 cross.
- the inclined surface 5 is not limited to two, and may be inclined by four surfaces such as a quadrangular pyramid, and the communication hole 15 may be provided at a position corresponding to the apex thereof. Or you may comprise a part as a slope in groove shape. In short, any slope may be used as long as the liquid 19 stored in the reservoir 2 can naturally flow toward the communication hole 15, and the slope includes a bent surface and a bottom surface provided with a groove.
- a plurality of communication holes 15 may be provided.
- the reaction unit 8 is recessed so that the lower surface opens on the lower surface side of the main body 30.
- a closed space (reaction chamber 17 described later) is formed.
- the slide glass is a glass plate used for placing a minute sample mainly during observation using an optical microscope.
- a slide glass having a general size of a short side of about 2.5 cm, a long side of about 7.5 cm, and a thickness of about 1.2 mm is employed.
- the illustrated reaction unit 8 has a square shape when viewed from the bottom surface, but is not limited thereto, and the shape viewed from the bottom surface may be a polygon, a circle, or an ellipse.
- the reaction part 8 is preferably a cylindrical space.
- the depth of the reaction part 8 is basically determined by the amount of the liquid 19 necessary for the reaction, but the liquid 19 that has flowed into the space surrounded by the reaction part 8 and the slide glass 16 (reaction chamber 17)
- the depth is preferably such that a force due to capillary action is generated, for example, a depth of 10 to 200 ⁇ m, more preferably 10 to 100 ⁇ m.
- the width and depth of the reaction part 8 (area of the opening 9) are set to be slightly smaller than the area of the lower surface of the rectangular parallelepiped while securing the thickness of the wall 10 surrounding the four sides of the reaction part 8.
- the capacity of the reaction unit 8 is different depending on the amount of the liquid 19 necessary for the staining reaction, similarly to the storage unit 2, and is not limited thereto, and can be changed as appropriate.
- the inner upper surface of the reaction unit 8 is composed of two inclined surfaces 11 and 11 that rise from the center toward the short sides 13 and 13.
- the magnitude of the inclination is preferably set to such a magnitude that the liquid 19 can reach the end portions 13 and 13 on the short side from the center by force due to capillary action. 1/100, more preferably substantially 1/200.
- the inclined surface 11 is not limited to two, like the inclined surface 5 of the storage unit 2, but may be four surfaces, or a part of the groove shape may be inclined, for example, if air or bubbles can move naturally.
- Such an inclined surface may be used, and the inclined surface includes a bent surface and a bottom surface provided with a groove.
- reaction unit 8 When the reaction unit 8 is placed and fixed on the slide glass 16, it constitutes a closed space (reaction chamber 17), so that the liquid 19 is difficult to dry. On the other hand, even if the amount of the liquid 19 supplied to the storage unit 2 is larger than the capacity of the reaction unit 8, the liquid 19 is difficult to leak because it is closed (at least during the time required for the reaction) it can.).
- a seal member, packing, or the like may be provided below the rectangular wall 10.
- a vent hole 14 penetrating from the inner upper surface of the reaction portion 8 to the upper surface of the main body 30 is formed. In the example of FIGS. 1 to 3, a total of four locations are provided, two at the end of each short side end portion 13. Note that the vent hole may communicate with an opening provided on the side surface of the main body 30.
- the number of the vent holes 14 is not limited to a mode in which two locations are provided at each short side end, and may be one location or three or more locations.
- the storage unit 2 and the reaction unit 8 are fluidly communicated with each other through a communication hole 15 provided in the center. More specifically, the communication hole 15 is formed so as to penetrate from the center of the bottommost part 6 on the inner and lower surfaces of the storage part 2 to the center of the inner upper surface of the reaction part 8. Since the communication hole 15 is located at the center of the reaction unit 8, the distance from the communication hole 15 to the end of the reaction unit 8 is shortened, and the liquid 19 is filled to every corner of the reaction unit 8 only by the force of capillary action. it can. Further, the liquid 19 stored in the storage unit 2 is transferred to the reaction unit 8 through the communication hole 15 by a certain amount due to its own weight and atmospheric pressure and the inclination of the inner and lower surfaces of the storage unit. Can be poured and no additional mechanism is required. Note that the speed (flow rate) at which the liquid 19 flows from the storage unit 2 into the reaction unit 8 can be controlled by changing the diameter and length of the communication hole 15.
- the liquid filling aid 1 of this embodiment may be formed of a metal material having a high thermal conductivity such as stainless steel. It is because the liquid 19 stored in the storage part 2 at the time of a heating can also be heated by doing so. Conversely, when it is desired to suppress the evaporation of the liquid 19 or when heating is not performed, the liquid 19 may be formed of a resin material having a low thermal conductivity. By doing so, even a small amount of the liquid 19 can be reacted. And if it is a transparent resin material, what can see the inside of the reaction part 8 can also be formed.
- FIG. 4 is a cross-sectional view illustrating the state of the liquid when the liquid filling aid of the present embodiment is installed on the slide glass and the liquid is discharged.
- (A) is a state immediately after the liquid is discharged to the storage part
- (b) is a state in which the liquid flows from the storage part through the communication hole to the reaction part, and (c) has been filled with the liquid in the entire reaction part.
- Each state is shown.
- the black arrow represents the liquid flow
- the white arrow represents the air flow.
- the liquid filling auxiliary tool 1 of the present embodiment is placed on the slide glass 16, and the space surrounded by the reaction unit 8 and the slide glass 16 forms a reaction chamber 17 in which a target substance reacts. It is preferable that the placed liquid filling auxiliary tool 1 is pressed downward by a holding tool such as an arm. In order to fill the reaction chamber 17 with the liquid 19 and cause the target substance to react, the following is performed.
- the liquid 19 is discharged from the discharge device 18 to the storage unit 2 (FIG. 4A). At this time, not only the capacity of the reaction chamber 17 but also the necessary amount is discharged to the storage unit 2 at once.
- the amount of the liquid 19 discharged to the storage unit 2 is preferably several times to several tens of times the capacity of the reaction chamber 17.
- the opening 3 of the storage unit 2 is formed to have a width that occupies half or more of the area of the upper surface of the main body 30, the liquid 19 can be discharged and supplied without precisely positioning the discharge device 18.
- the liquid 19 supplied to the storage unit 2 is moved to the upper part of the communication hole 15 at the center along the inclined surface 5 at the bottom of the storage unit 2 by the action of the weight and atmospheric pressure and the inclined surface 5 at the bottom of the storage unit 2. And flows into the communication hole 15 as it is (reference numeral 20). Then, the liquid 19 that has flowed into the reaction chamber 17 through the communication hole 15 is caused by the force of capillary action generated by the space sandwiched between the upper surface 11 of the reaction chamber 17 and the slide glass 16, and the short side on the left and right sides. It flows toward the end portions 13 and 13 (reference numeral 21).
- the liquid 19 stored in the storage unit 2 is larger than the capacity of the reaction chamber 17, the liquid 19 continues to flow into the reaction chamber 17 and reaches the short side end portions 13, 13 of the reaction chamber 17. It goes up in 14 each.
- the liquid 19 rising in the four vent holes 14 stops when it balances with the liquid 19 in the reservoir 2 (FIG. 4C). This is because the reservoir 2 is provided in fluid communication with the communication hole 15 directly above the reaction chamber 17, and the vent hole 14 is parallel to the reservoir 2 and is the same as the upper end of the reservoir 2 from the reaction chamber 17. This is because it penetrates to the height.
- the liquid 19 rising in the vent holes 14 and the liquid 19 remaining in the reservoir 2 are replenished when the liquid 19 in the reaction chamber 17 is reduced instead of additional discharge. Moreover, by raising the inside of the vent hole 14, the contact area between the liquid 19 and the outside air can be reduced, and unnecessary drying can be prevented.
- the gap between the reaction chamber 17 and the slide glass 16 is a sufficient gap from the center to the end portions 13 and 13 on the short side to cause force due to capillary action, and the upper surface of the reaction chamber 17. 11 and 11 are inclined, and a plurality of vent holes 14 are provided at the highest position 12 of the inclination. Therefore, the entire reaction chamber 17 can be filled with the liquid 19 without mixing bubbles.
- the storage unit 2 having a larger capacity than the reaction chamber 17 is provided immediately above the reaction chamber 17 and the reaction chamber 17 and the storage unit 2 are connected by the communication hole 15, a necessary amount of the liquid 19 is stored. This eliminates the need for additional discharge of the liquid 19.
- the liquid filling auxiliary tool 1 of the second embodiment is mainly different from the first embodiment in that it includes a deep digging portion 24. Below, it demonstrates centering on the point which is different from 1st Embodiment, and omits description about a common point.
- 8 is a top view of the liquid filling aid 1 of the present embodiment
- FIG. 9A is a cross-sectional view taken along line BB in FIG. 8
- FIG. 9B is a cross-sectional view taken along line CC in FIG. Sectional drawing shown by is shown.
- the liquid filling auxiliary tool 1 of the present embodiment includes a reservoir 30, a reaction unit 8, a vent hole 14, and a communication hole 15 in a main body 30 that is a rectangular parallelepiped member, and is installed on a slide glass 16.
- the points used are the same as in the first embodiment.
- the liquid filling auxiliary tool 1 of the present embodiment is different from the first embodiment in that the deep digging portion 24 is formed at the center of the storage portion 2.
- the deep digging portion 24 is a groove provided in the center of the storage portion 2 and has an elongated oval shape extending in the short side direction of the storage portion 2 when viewed from above.
- a mortar-shaped inclined surface 25 is formed in the center direction, and a communication hole 15 is provided at the center of the inclined surface 25. Since the inclined surface 25 is steeper than the inclined surface 5, the residual liquid on the inclined surface 25 is minimal.
- the shapes of the deep excavation part 24 and the inclined surface 25 are symmetrical with respect to the center lines in the vertical direction and the horizontal direction.
- the shape of the deep digging portion 24 is not limited to the illustrated elliptical shape, and the shape of the inclined surface 25 is not limited to the mortar shape.
- the volume of the deep digging portion 24 is such that when the deep digging portion 24 is called a first concave portion and the storage portion 2 excluding the deep digging portion 24 is called a second concave portion, the volume of the second concave portion is equal to the volume of the first concave portion. It is configured to be 2 times or more, preferably 10 times or more, and more preferably 20 times or more.
- the volume of the 2nd crevice (storage part 2 excluding deep digging part 24) of a 2nd embodiment is 100 times or more of the 1st crevice (deep digging part 24). From another viewpoint, the volume of the deep section 24 is substantially equal to or less than the volume of the reaction chamber 17, preferably 1 ⁇ 2 or less of the volume of the reaction chamber 17, more preferably the volume of the reaction chamber 17. 1/3 or less.
- the technical significance of the formation of the deep excavation portion 24 is that a minute amount (for example, several hundred ⁇ L) of an expensive liquid used in a certain step of the immunohistochemical staining method is reliably transferred from the communication hole 15 to the reaction portion 8. It is to be able to supply.
- a minute amount of liquid for example, an anti-human HER2 / neu gene product polyclonal antibody used as a primary antibody in the fourth step described in paragraph 0002 of the present specification is disclosed.
- an amount of liquid ranging from substantially the same amount as the volume of the deep digging portion 24 to several times the amount is discharged from the discharge device to the approximate center of the storage portion 2. Supplied to the reservoir 2.
- the liquid filling aid 1 is used. It must be possible.
- the liquid filling auxiliary tool 1 of the present embodiment includes the second concave portion (the storage portion 2 excluding the deep digging portion 24) having a larger volume than the first concave portion (the deep digging portion 24).
- the liquid filling auxiliary tool 1 of the present embodiment includes the second concave portion (the storage portion 2 excluding the deep digging portion 24) having a larger volume than the first concave portion (the deep digging portion 24).
- an antigen activation solution (10 mM sodium citrate buffer pH 6.0) used in the third step described in paragraph 0002 of the present specification is disclosed.
- the step of using a small amount of liquid and the step of using a relatively large amount of liquid in the immunohistochemical staining method are performed by the same liquid filling aid 1. It becomes possible to carry out.
- FIG. 5 is a block diagram illustrating the automatic staining apparatus 51 of the embodiment.
- the automatic dyeing device 51 of this embodiment includes a discharge device 18, a drive device 52, an elevating device 53, an arm (holding tool) 54, a stage 55, a heater 56, and a control device 61.
- the discharge device 18 is a device that discharges a liquid 19 for reacting a biological tissue or a cell adhered or pasted on the slide glass 16.
- the discharge device 18 is in close contact with the inner surface of a storage container having a nozzle at the tip.
- a plunger type discharge device that moves and moves a moving plunger by a desired amount is used.
- the plunger type discharge device is used, but it is of course possible to use other types of discharge devices.
- an air type that applies pressure-adjusted air to a liquid in a storage container having a nozzle at the tip for a desired time, or a tube that operates by crushing a plurality of rollers or a keyboard-like member against a flexible tube
- a tubing type or the like for transferring the liquid inside can be used.
- the supply of the liquid 19 to the discharge device 18 is normally performed by a storage container provided in the discharge device 18, but when using a plunger type, the liquid 19 is prepared in another container, and before the discharge, Suction may be performed.
- the discharge device 18 is installed in the drive device 52 and is movable in the XYZ directions (reference numeral 63).
- the driving device 52 for example, a device driven by a combination of a ball screw and a servo motor or a stepping motor, a device driven by a linear motor, or the like can be used.
- the driving device 52 may be realized by a plurality of driving devices that allow the ejection device 18 and the liquid filling auxiliary tool 1 to move relative to each other in the XYZ directions. This may be realized by providing a first drive device for moving in the X direction and a second drive device for moving in the X direction, Y direction, and Z direction on the ejection device 18 side.
- staining apparatus 51 of a present Example is equipped with the raising / lowering apparatus 53 in order to mount the liquid filling auxiliary tool 1 of embodiment on the slide glass 16, and to fix it.
- the liquid filling auxiliary tool 1 is gripped by an arm 54 attached to the lifting device 53, and moves up and down between a rising position separated from the stage 55 and a lowering position where the stage 55 and the slide glass 16 are in contact with each other (see FIG. 62).
- the lifting device 53 for example, the same device as the driving device 52, an air cylinder, or the like can be used.
- the arm 54 may be provided with a rotation shaft for inclining the grasped liquid filling aid 1 to discharge the liquid 19.
- the lower side of the slide glass 16 is provided with a frame-shaped pallet 57 that holds the slide glass 16 and a stage 55 that supports the slide glass 16 that is pressed in a central opening of the pallet 57.
- a groove-like portion that is substantially the same as or slightly larger than the size of the slide glass 16 is recessed.
- a protrusion having a height not exceeding the thickness of the slide glass 16 may be provided on the upper surface of the pallet 57 so as to surround the slide glass 16.
- the stage 55 includes a heater 56 and can heat the slide glass 16 in contact with the stage 55.
- a liquid receiving container 58 and a tank 59 that is connected to the liquid receiving container 58 and stores various liquids received by the liquid receiving container 58 are provided below the stage 55.
- the liquid receiving container 58 is used when recovering the cleaning liquid after the cleaning in the cleaning process described later, the liquid 19 after the reaction is completed, and the like.
- the pallet 57 and the stage 55 have the following relationship.
- the pallet 57 is supported from the lower surface by a spring 64, and the surface (upper surface) for holding the slide glass 16 before the liquid filling auxiliary tool 1 is placed is the surface for supporting the slide glass 16 of the stage 55 ( It is above the upper surface (FIG. 6A).
- the spring 64 supporting the pallet 57 is contracted and the lower surface of the slide glass 16 is positioned within the opening of the pallet 57.
- the glass slide 16 is brought into contact with the upper surface of the stage 55 and the liquid filling auxiliary tool 1 and the stage 55 are clamped and fixed (FIG. 6B).
- the slide glass 16 and the liquid filling auxiliary tool 1 are fixed without a gap by the downward pressing force of the lifting / lowering device 53 and the upward biasing force of the spring 64, and even if no seal member / packing is provided, leakage occurs. It is possible to form the reaction chamber 17 with a small amount. Further, since the slide glass 16 is firmly fixed to the stage 55, when the heater 56 is used, heat can be efficiently transmitted.
- the automatic staining apparatus 51 of the present embodiment includes an air blowing device 60 in order to remove dust, cleaning liquid, and the like attached on the slide glass 16.
- the air blow device 60 includes a nozzle that is connected to a compressed air source and blows out the compressed air, and uses a device that switches supply and stop of the compressed air by an electromagnetic valve or the like. Other configurations may be used as long as the compressed air can be selectively blown.
- Each device described above is controlled by the control device 61.
- the control device 61 is mainly composed of an input / output device, a storage device, and a processing device. For example, a personal computer or a programmable controller can be used.
- FIG. 7 shows a flowchart for explaining an example of the operation of the automatic staining apparatus of the embodiment.
- the operation of the automatic staining apparatus shown below is a representative one of a plurality of steps constituting the above-described immunohistochemical staining operation (corresponding to the second and third steps in the above-described steps).
- movement in is illustrated.
- the contents may be slightly different, such as the presence or absence of stage heating, the presence or absence of raising and lowering the liquid filling aid, and the reaction waiting time.
- the slide glass 16 to which the target substance is bonded is placed on the pallet 57 (STEP 101). Then, air blow is performed to remove dust and cleaning liquid adhering to the slide glass 16 (STEP 102).
- the liquid filling auxiliary tool 1 is lowered by the lifting device 53, and the slide glass 16 is fixed so as to be sandwiched between the liquid filling auxiliary tool 1 and the stage 55 (STEP 103).
- heating is performed after fixing the slide glass 16, heating of the stage 55 is started (STEP 104). When not heating, it progresses to the next step, without performing STEP104. And the liquid 19 is discharged toward the storage part 2 of the liquid filling auxiliary tool 1 upper part (STEP 105).
- the liquid 19 discharged to the reservoir 2 at the upper part of the liquid filling aid 1 flows into the reaction part 8 through the communication hole 15 and is surrounded by the slide glass 16 and the reaction part 8. Filled. Then, it reacts with the target substance that has been bonded to the slide glass 16 in advance. It waits until predetermined time passes so that this reaction may fully be performed (STEP106). This time varies depending on the target substance and the type of liquid. When the predetermined time has elapsed, if the heating has been performed, the heating of the stage 55 is terminated (STEP 107). If the heating has not been performed, the process proceeds to the next step without executing STEP 107.
- the liquid filling auxiliary tool 1 is not raised, and the cleaning liquid is discharged as it is, and the reaction chamber 17 is filled in the same manner as in the case of the liquid 19 (STEP 108).
- the liquid filling aid 1 is raised and the cleaning liquid is discharged (STEP 109).
- the discharged cleaning liquid is collected in the liquid receiving container 58 and stored in the tank 59.
- air blow is performed to remove the cleaning liquid remaining on the slide glass 16 (STEP 110).
- the above is a typical one-step operation.
- step (STEP 101) of placing the slide glass 16 on the pallet 57 or the air blow step (STEP 102 or STEP 110) may be omitted.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Dispersion Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
Description
免疫組織化学染色の操作は、一般的に、次のような複数の工程から構成される。第1の工程は、スライドガラス上に貼付したホルマリン溶液などを用いて固定した検体を包埋しているパラフィンを有機溶媒にて溶解する脱パラフィン工程、第2の工程は、有機溶媒を取り除き、水になじませるための親水化工程、第3の工程は、免疫反応性を高めるための処理を行う抗原賦活化工程、第4の工程は、検体中の抗原と反応する抗体を適用する一次抗体適用工程、第5の工程は、反応した抗体を検出するための試薬を適用する検出工程、第6の工程は、検出試薬を可視化するための試薬を適用する発色工程、第7の工程は、組織の形状を見やすくするために、核を異なる色に染色する核染色工程である。
(1)カバー短辺側からもう一方の短辺側まで比較的長い距離(スライドガラス長手方向のほぼ全長)にわたって充填を行うにあたり、毛細管現象による力のみでは、充填が不十分になる虞がある。そのため、カバーを滑動したり、吐出側とは反対側から吸引をしたりするなど、充填を補助する付加的な機構が必要である。なお、上記特許文献3の形状では、そもそも毛管作用によりカバーと基部とで囲まれた空間全体を液体で満たすことはできない。
(2)カバー短辺側端部は広く開口しているので、そこから液体の蒸発による乾燥が起こりやすく、乾燥が起きると、乾燥部分に抗原抗体反応によらない非特異染色が起きる虞がある。そのため、充分反応が進行するまで、追加の液体充填が必要になるという課題がある。
(3)温めて反応させる方法の場合には、液体の蒸発が促進されて乾燥してしまうので、反応不足や染色むらが起きる虞がある。そのため、充分反応が進行するまでは、追加の液体充填が必要になるという課題がある。
しかし、温度の低い液体を追加すること(換言すると、すでに温まっている液体とこれから温める液体とを混ぜること)は、反応相の温度を低下させることになり、十分な熱量を組織に与えることができない。かかる場合、染色むらや染色の不足が起きる可能性があり、反応にとって好ましくないため、液体の温度管理(温調)が必要になる。
(5)カバーの端部などの狭い範囲に吐出を行わなければならないので、吐出口の精密な位置決めが必要である。
第2の発明は、第1の発明において、前記貯留部(2)の容積が反応部(8)の容積より大きいことを特徴とする。
第3の発明は、第1の発明において、前記貯留部(2)が、連通孔を挟んで反応部(8)の上方に配置されることを特徴とする。
第4の発明は、第3の発明において、前記本体(30)の上面に貯留部の開口(3)が設けられていることを特徴とする。ここで、前記連通孔(15)が、反応部(8)の略中心に位置することを特徴としてもよい。
第5の発明は、第1の発明において、前記通気孔(14)が、反応部(8)の上面から本体(30)の上面まで貫通していることを特徴とする。
第6の発明は、第1の発明において、前記反応部(8)が、反応部の全域において毛細管現象による力が発生する深さに構成されることを特徴とする。
第7の発明は、第6の発明において、前記深さが、10~200μmであることを特徴とする。
第8の発明は、第1の発明において、前記貯留部(2)の底面が連通孔に向かう傾斜面または溝を有することを特徴とする。
第9の発明は、第1の発明において、前記連通孔(15)に向かう傾斜面の少なくとも1つが、2つ以上の傾斜を有することを特徴とする。
第10の発明は、第1の発明において、前記貯留部(2)が、前記連通孔が略中心に設けられた第一凹部(24)と、第一凹部よりも上方に位置しかつ容積が2倍以上大きい第二凹部と、を備えて構成されることを特徴とする。
第11の発明は、第1の発明において、前記反応部(8)の内上面(天井面)が通気孔に向かう傾斜面または溝を有し、前記通気孔(14)が、反応部の内上面の最高位置に設けられていることを特徴とする。
第12の発明は、第1ないし11のいずれかの発明において、前記本体(30)が、金属材料からなることを特徴とする。
第13の発明は、第1ないし11のいずれかの発明において、前記板状体が、染色工程で用いられるスライドガラスであることを特徴とする。
第15の発明は、第1ないし11のいずれかの発明に係る液体充填補助具を用いた液体充填方法であって、液体充填補助具を板状体の上に設置するステップと、反応室の容積以上の量の液体を貯留部へ吐出するステップとを含むことを特徴とする液体充填方法である。
(1)反応部と貯留部を連通する連通孔を備えるので、毛細管現象による力のみでも反応部の隅々まで液体を充填できる。また、貯留部に溜めた液体は連通孔から反応部へと自重で流下するから、充填を補助する付加的な機構は必要ない。
(2)スライドガラスに本液体充填補助具を設置したとき、反応部は閉じた空間である反応室を構成するので、乾燥を防止できる。また、通気孔に達するまで反応部内に液体を充填することで、液体と空気の接触面積を減らし、乾燥を遅らせることができる。
(3)液体を溜める貯留部があるので、液体の蒸発の問題にも対応することができる。また、液体を溜める貯留部が反応部に隣接して設けられているので、反応のための加熱とともに、貯留部に溜めた液体も加熱することができる。
(4)反応部と連通する通気孔を設けているので、気泡が混入したとしても、通気孔より排出される。
(5)貯留部上面が広く開口する構成においては、吐出口の精密な位置決めが不要である。
《第1実施形態》
<構成>
図1に本実施形態の液体充填補助具1の上面図、図2に図1内にA-A線で示した断面図、図3に本実施形態の液体充填補助具1の下面図を示す。以下では、図1或いは図3の左右方向を幅方向、上下方向を奥行き方向、図2の上下方向を高さ方向と呼ぶことがある。
本実施形態の液体充填補助具1は、直方体状の部材である本体30に、貯留部2と、反応部8と、通気孔14と、連通孔15とを形成して構成され、後述するスライドガラス16に設置して使用される。ここで、液体充填補助具1の奥行き方向の長さは、スライドガラス16の短辺の長さと同じかやや短くする。また、幅方向の長さは、スライドガラス16の長辺の長さの約2/3とする。この長さは、スライドガラス16の使用可能領域に対応して適宜変更するものである。なお、液体充填補助具1に覆われないスライドガラス16の残り約1/3の部分には、識別のための文字や記号、バーコードなどが記されるスペースとなる。本体30の形状は、例示の直方体状に限定されず、多角形柱、円柱または楕円柱であってもよい。以下に、各構成について詳説する。
反応部8の傾斜面11の最高位置12には、反応部8内上面から本体30の上面まで貫通する通気孔14を穿設する。図1~3の例では、各短辺側端部13の端の二箇所ずつ計四箇所設けている。なお、通気孔は、本体30の側面に設けられた開口に連通するようにしてもよい。
図4に本実施形態の液体充填補助具をスライドガラスに設置し、液体を吐出したときの液体の状態を説明する断面図を示す。(a)は貯留部に液体を吐出した直後の状態、(b)は貯留部から連通孔を通って反応部へと液体が流れ込む状態、(c)は反応部全体に液体を充填し終えた状態をそれぞれ示す。図中、黒塗り矢印は液体の流れを、白塗り矢印は空気の流れを表す。
本実施形態の液体充填補助具1をスライドガラス16の上に載置し、反応部8とスライドガラス16とで囲まれた空間が、対象となる物質を反応させる反応室17を形成する。載置した液体充填補助具1は、アーム等の保持具で下方に押さえつけることが好ましい。この反応室17に液体19を満たし、対象となる物質を反応させるには、次のようにする。
貯留部2に供給された液体19は、自重および大気圧並びに貯留部2の底部の傾斜面5の作用により、貯留部2の底部の傾斜面5に沿って中央にある連通孔15の上部へと導かれ、そのまま連通孔15へと流れ込む(符号20)。そして、連通孔15を通って反応室17へと流れ込んだ液体19は、反応室17の上面11とスライドガラス16とで挟まれた空間により生起される毛細管現象による力により、左右の短辺側端部13、13へと向かって流れていく(符号21)。そのとき、反応室17内にあった空気は、液体19よりも軽いため、反応室17上面11の傾斜に沿って進み、傾斜の最高位置12にある通気孔14を通って外部へと排出される(符号22)(図4(b))。
第2実施形態の液体充填補助具1は、深掘部24を備えている点で主として第1実施形態と相違する。以下では、第1実施形態と相違する点を中心に説明し、共通する点については説明を割愛する。
図8に本実施形態の液体充填補助具1の上面図、図9(a)に図8内にB-B線で示した断面図、図9(b)に図8内にC-C線で示した断面図を示す。
本実施形態の液体充填補助具1は、直方体状の部材である本体30に、貯留部2と、反応部8と、通気孔14と、連通孔15とを備え、スライドガラス16に設置して使用される点は第1実施形態と同様である。しかし、本実施形態の液体充填補助具1は、貯留部2の中央に深掘部24が形成されている点で第1実施形態と相違する。
深掘部24の容積は、深掘部24を第一凹部と呼称し、深掘部24を除く貯留部2を第二凹部と呼称した場合、第二凹部の容積が第一凹部の容積の2倍以上、好ましくは10倍以上、さらに好ましくは20倍以上となるように構成する。第2実施形態の第二凹部(深掘部24を除く貯留部2)の容積は、第一凹部(深掘部24)の100倍以上となっている。別の観点からは、深掘部24の容積は反応室17の容積と実質的に同じかそれ以下であり、好ましくは反応室17の容積の1/2以下、より好ましくは反応室17の容積の1/3以下とする。
図5に実施例の自動染色装置51を説明するブロック図を示す。
本実施例の自動染色装置51は、吐出装置18と、駆動装置52と、昇降装置53と、アーム(保持具)54と、ステージ55と、ヒーター56と、制御装置61とを主要な構成要素とする。
吐出装置18は、スライドガラス16上に接着ないし貼付された生物組織や細胞などを反応させるための液体19を吐出する装置であり、本実施例では先端にノズルを有する貯留容器の内面に密着摺動するプランジャを所望量移動して吐出するプランジャ式吐出装置を用いる。本実施例ではプランジャ式吐出装置を用いたが、これ以外の方式の吐出装置を用いることももちろん可能である。例えば、先端にノズルを有する貯留容器内の液体に調圧されたエアを所望時間だけ印加するエア式や、複数のローラーや鍵盤状部材を可撓性チューブに対して押しつぶすように動作させてチューブ内の液体を移送するチュービング式などを用いることができる。吐出装置18への液体19の供給は、吐出装置18が備える貯留容器により行われるのが通常であるが、プランジャ式を用いる場合は、液体19を別の容器に用意しておき、吐出前に吸引を行うようにしてもよい。
また、本実施例の自動染色装置51は、実施形態の液体充填補助具1をスライドガラス16上に載置し、固定するために昇降装置53を備える。液体充填補助具1は、昇降装置53に取り付けられたアーム54により把持し、ステージ55から離間した上昇位置と、ステージ55とスライドガラス16を挟んで接触する下降位置との間で昇降動する(符号62)。昇降装置53としては、例えば、上記駆動装置52と同様のものや、エアシリンダなどを用いることができる。アーム54には、把持した液体充填補助具1を傾けて液体19を排出するための回動軸を設けてもよい。
上述した各機器は、制御装置61により制御される。制御装置61は、入出力装置、記憶装置、処理装置から主に構成される。例えば、パーソナルコンピュータやプログラマブルコントローラなどを用いることができる。
図7に実施例の自動染色装置の動作の例を説明するフローチャートを示す。
以下に示す自動染色装置の動作は、前述した免疫組織化学染色の操作を構成する複数の工程のうち、代表的な一工程(前述した工程でいえば第2、第3の工程に相当する)における動作を例示するものである。ただし、工程の種類によっては、ステージ加熱の有無や液体充填補助具昇降の有無、反応待ち時間など、内容が若干異なる場合がある。
Claims (15)
- 板状体の上に設置され、液体が充填される反応室を構成する液体充填補助具であって、
本体と、
本体に形成された液体を貯留するための貯留部と、
本体の下面に設けられた凹みである反応部と、
貯留部と反応部とを流体的に連通する連通孔と、
反応部と大気を連通する通気孔とを備え、
反応部および板状体の上面が、反応室を構成することを特徴とする液体充填補助具。 - 前記貯留部の容積が反応部の容積より大きいことを特徴とする請求項1の液体充填補助具。
- 前記貯留部が、連通孔を挟んで反応部の上方に配置されることを特徴とする請求項1の液体充填補助具。
- 前記本体の上面に貯留部の開口が設けられていることを特徴とする請求項3の液体充填補助具。
- 前記通気孔が、反応部上面から本体の上面まで貫通していることを特徴とする請求項1の液体充填補助具。
- 前記反応部が、反応部の全域において毛細管現象による力が発生する深さに構成されることを特徴とする請求項1の液体充填補助具。
- 前記深さが、10~200μmであることを特徴とする請求項6の液体充填補助具。
- 前記貯留部の底面が連通孔に向かう傾斜面または溝を有することを特徴とする請求項1の液体充填補助具。
- 前記連通孔に向かう傾斜面の少なくとも1つが、2つ以上の傾斜を有することを特徴とする請求項8の液体充填補助具。
- 前記貯留部が、前記連通孔が設けられた第一凹部と、第一凹部よりも上方に位置しかつ容積が2倍以上大きい第二凹部と、を備えて構成されることを特徴とする請求項1の液体充填補助具。
- 前記反応部の内上面(天井面)が通気孔に向かう傾斜面または溝を有し、前記通気孔が、反応部の内上面の最高位置に設けられていることを特徴とする請求項1の液体充填補助具。
- 前記本体が、金属材料からなることを特徴とする請求項1ないし11のいずれかの液体充填補助具。
- 前記板状体が、染色工程で用いられるスライドガラスであることを特徴とする請求項1ないし11のいずれかの液体充填補助具。
- 液体の吐出装置と、
スライドガラスを保持するパレットと、
吐出装置をパレットに対しXYZ方向に相対的に移動自在とする駆動装置と、
請求項13の液体充填補助具を保持する保持具と、
保持具を昇降動作させる昇降装置と、を備える自動染色装置。 - 請求項1ないし11のいずれかの液体充填補助具を用いた液体充填方法であって、
液体充填補助具を板状体の上に設置するステップと、
反応室の容積以上の量の液体を貯留部へ吐出するステップとを含むことを特徴とする液体充填方法。
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP14737779.0A EP2944940B1 (en) | 2013-01-08 | 2014-01-08 | Aid for filling liquid, and method for filling liquid |
US14/759,612 US9719896B2 (en) | 2013-01-08 | 2014-01-08 | Aid for filling liquid, and method for filling liquid |
CN201480004310.9A CN104903696B (zh) | 2013-01-08 | 2014-01-08 | 液体填充辅助用具以及液体填充方法 |
KR1020157020605A KR102198627B1 (ko) | 2013-01-08 | 2014-01-08 | 액체 충전 보조구 및 액체 충전 방법 |
JP2014556421A JP6318093B2 (ja) | 2013-01-08 | 2014-01-08 | 液体充填補助具および液体充填方法 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2013001023 | 2013-01-08 | ||
JP2013-001023 | 2013-01-08 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2014109323A1 true WO2014109323A1 (ja) | 2014-07-17 |
Family
ID=51166971
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2014/050100 WO2014109323A1 (ja) | 2013-01-08 | 2014-01-08 | 液体充填補助具および液体充填方法 |
Country Status (6)
Country | Link |
---|---|
US (1) | US9719896B2 (ja) |
EP (1) | EP2944940B1 (ja) |
JP (1) | JP6318093B2 (ja) |
KR (1) | KR102198627B1 (ja) |
CN (1) | CN104903696B (ja) |
WO (1) | WO2014109323A1 (ja) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US12005441B1 (en) | 2014-11-26 | 2024-06-11 | Medica Corporation | Automated microscopic cell analysis |
US11478789B2 (en) | 2014-11-26 | 2022-10-25 | Medica Corporation | Automated microscopic cell analysis |
US20170328924A1 (en) | 2014-11-26 | 2017-11-16 | Ronald Jones | Automated microscopic cell analysis |
US10625259B1 (en) | 2014-11-26 | 2020-04-21 | Medica Corporation | Automated microscopic cell analysis |
EP3289366B1 (en) | 2015-05-01 | 2021-12-29 | Abbott Laboratories | Apparatus for removing liquid contents of a container |
US9976946B2 (en) * | 2016-04-21 | 2018-05-22 | Instrumentation Laboratory Company | Optical flow cell and test head apparatus |
US11047845B1 (en) | 2017-11-15 | 2021-06-29 | Medica Corporation | Control material and methods for cell analyzers |
CN113834715B (zh) * | 2020-06-23 | 2024-05-14 | 上海伯顿医疗设备有限公司 | 一种多玻片装载量、气动雾化喷液的染色机 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0514891U (ja) * | 1991-03-22 | 1993-02-26 | 澄晴 野地 | スライドグラス用ホルダ |
JP2000508423A (ja) | 1996-04-12 | 2000-07-04 | ヴィジョン インストゥルメンツ リミテッド | ヒト又は動物の細胞試料の処理方法及び処理装置 |
JP2002350305A (ja) * | 2001-05-28 | 2002-12-04 | Aloka Co Ltd | 液展開用部材及びそれを用いたサンプル処理装置 |
JP2005530208A (ja) | 2002-06-20 | 2005-10-06 | ビジョン・バイオシステムズ・リミテッド | 一体化された容器を備えた顕微鏡スライドカバー |
JP2007532918A (ja) | 2004-04-16 | 2007-11-15 | チュー,ウェイシン | 組織標本からの生体分子抽出装置 |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4392450A (en) * | 1981-02-12 | 1983-07-12 | Prevo Donald L | Device for spreading monolayered films |
GB8722902D0 (en) | 1987-09-30 | 1987-11-04 | Shandon Southern Prod | Tissue &c processing |
US5922604A (en) * | 1997-06-05 | 1999-07-13 | Gene Tec Corporation | Thin reaction chambers for containing and handling liquid microvolumes |
US6673620B1 (en) * | 1999-04-20 | 2004-01-06 | Cytologix Corporation | Fluid exchange in a chamber on a microscope slide |
ATE416369T1 (de) * | 1999-04-20 | 2008-12-15 | Cytologix Corp | Flüssigkeitswechsel in einer kammer auf einem mikroskopobjektträger |
US7468161B2 (en) | 2002-04-15 | 2008-12-23 | Ventana Medical Systems, Inc. | Automated high volume slide processing system |
ES2424988T3 (es) * | 2002-04-15 | 2013-10-10 | Ventana Medical Systems, Inc. | Sistema automatizado de coloración de placas portaobjetos con elevada velocidad de producción |
US20090286305A1 (en) | 2005-04-15 | 2009-11-19 | Wei-Sing Chu | Method for non-destructive macromolecule extraction from biological samples on slide |
JP5881936B2 (ja) * | 2009-04-20 | 2016-03-09 | ソニー株式会社 | 試料溶液導入キット及び試料溶液注入器 |
-
2014
- 2014-01-08 WO PCT/JP2014/050100 patent/WO2014109323A1/ja active Application Filing
- 2014-01-08 KR KR1020157020605A patent/KR102198627B1/ko active IP Right Grant
- 2014-01-08 US US14/759,612 patent/US9719896B2/en active Active
- 2014-01-08 JP JP2014556421A patent/JP6318093B2/ja active Active
- 2014-01-08 EP EP14737779.0A patent/EP2944940B1/en active Active
- 2014-01-08 CN CN201480004310.9A patent/CN104903696B/zh active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0514891U (ja) * | 1991-03-22 | 1993-02-26 | 澄晴 野地 | スライドグラス用ホルダ |
JP2000508423A (ja) | 1996-04-12 | 2000-07-04 | ヴィジョン インストゥルメンツ リミテッド | ヒト又は動物の細胞試料の処理方法及び処理装置 |
JP2002350305A (ja) * | 2001-05-28 | 2002-12-04 | Aloka Co Ltd | 液展開用部材及びそれを用いたサンプル処理装置 |
JP2005530208A (ja) | 2002-06-20 | 2005-10-06 | ビジョン・バイオシステムズ・リミテッド | 一体化された容器を備えた顕微鏡スライドカバー |
JP2007532918A (ja) | 2004-04-16 | 2007-11-15 | チュー,ウェイシン | 組織標本からの生体分子抽出装置 |
Also Published As
Publication number | Publication date |
---|---|
EP2944940B1 (en) | 2020-07-08 |
JP6318093B2 (ja) | 2018-04-25 |
US9719896B2 (en) | 2017-08-01 |
EP2944940A4 (en) | 2016-09-07 |
US20160003718A1 (en) | 2016-01-07 |
JPWO2014109323A1 (ja) | 2017-01-19 |
KR20150103192A (ko) | 2015-09-09 |
CN104903696B (zh) | 2019-02-19 |
KR102198627B1 (ko) | 2021-01-05 |
CN104903696A (zh) | 2015-09-09 |
EP2944940A1 (en) | 2015-11-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6318093B2 (ja) | 液体充填補助具および液体充填方法 | |
US7635453B2 (en) | Device and method for wetting objects | |
CN102687023B (zh) | 用于可调容纳体积的薄膜处理装置 | |
US9207249B2 (en) | Automated system for handling microfluidic devices | |
JP5646196B2 (ja) | 吐出装置および液体分注装置並びに液体分注方法 | |
AU2006200551B2 (en) | Reagent container and slide reaction and retaining tray, and method of operation | |
JP2005536727A5 (ja) | ||
US8691163B2 (en) | Sample solution introduction kit and sample solution injector | |
US20090223012A1 (en) | Liquid suction device | |
CN116298353A (zh) | 一种用于微体积液体移液操作的装置 | |
EP2075586A1 (en) | Dispenser | |
US20140256598A1 (en) | Methods and systems for controlling liquids in multiplex assays | |
JP2016067277A (ja) | 生体関連物質抽出デバイス及び生体関連物質抽出装置 | |
JP4551123B2 (ja) | マイクロ流体システム及びそれを用いる処理方法 | |
JP2010025854A (ja) | マイクロ検査チップ | |
EP4056274A1 (en) | Structures for supporting the filling of wells in microfluidic devices | |
JP2018191608A (ja) | 核酸増幅装置及び核酸増幅方法 | |
CN115770627A (zh) | 一种液滴阅读芯片及其使用方法 | |
KR100743500B1 (ko) | 액체 렌즈의 전면 패널 합착 장치 및 방법 | |
JP2016198046A (ja) | 生体関連物質抽出デバイス、生体関連物質抽出装置、及び生体関連物質抽出容器 | |
JP4665416B2 (ja) | 液晶注入方法とその装置。 | |
KR101365609B1 (ko) | 접착액 도포 장치 및 이를 이용한 접착액 도포 방법 | |
JP2004157262A (ja) | 材料注入方法及び材料注入装置 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 14737779 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2014556421 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 14759612 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2014737779 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 20157020605 Country of ref document: KR Kind code of ref document: A |