JP6263647B2 - ヘマトキシリン沈殿物の洗浄方法及びシステム - Google Patents
ヘマトキシリン沈殿物の洗浄方法及びシステム Download PDFInfo
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- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 title claims description 74
- 239000002244 precipitate Substances 0.000 title claims description 44
- 238000000034 method Methods 0.000 title claims description 29
- 238000004140 cleaning Methods 0.000 title description 12
- 238000010186 staining Methods 0.000 claims description 31
- 239000007788 liquid Substances 0.000 claims description 29
- HLUCICHZHWJHLL-UHFFFAOYSA-N hematein Chemical compound C12=CC=C(O)C(O)=C2OCC2(O)C1=C1C=C(O)C(=O)C=C1C2 HLUCICHZHWJHLL-UHFFFAOYSA-N 0.000 claims description 26
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical group OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 20
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 239000013049 sediment Substances 0.000 claims description 13
- 230000001590 oxidative effect Effects 0.000 claims description 11
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 8
- 238000009826 distribution Methods 0.000 claims description 6
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- 239000001488 sodium phosphate Substances 0.000 claims 1
- WUWHFEHKUQVYLF-UHFFFAOYSA-M sodium;2-aminoacetate Chemical compound [Na+].NCC([O-])=O WUWHFEHKUQVYLF-UHFFFAOYSA-M 0.000 claims 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 claims 1
- 229910000406 trisodium phosphate Inorganic materials 0.000 claims 1
- 235000019801 trisodium phosphate Nutrition 0.000 claims 1
- 239000000243 solution Substances 0.000 description 44
- 238000002474 experimental method Methods 0.000 description 15
- 238000012360 testing method Methods 0.000 description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 238000004043 dyeing Methods 0.000 description 6
- 238000012545 processing Methods 0.000 description 6
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 5
- 241001510071 Pyrrhocoridae Species 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- 239000004744 fabric Substances 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 4
- 238000009825 accumulation Methods 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 3
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- 210000001519 tissue Anatomy 0.000 description 3
- 229920000858 Cyclodextrin Polymers 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- NALMPLUMOWIVJC-UHFFFAOYSA-N n,n,4-trimethylbenzeneamine oxide Chemical compound CC1=CC=C([N+](C)(C)[O-])C=C1 NALMPLUMOWIVJC-UHFFFAOYSA-N 0.000 description 2
- 235000006408 oxalic acid Nutrition 0.000 description 2
- 230000000704 physical effect Effects 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000011697 sodium iodate Substances 0.000 description 2
- 229940032753 sodium iodate Drugs 0.000 description 2
- 235000015281 sodium iodate Nutrition 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- LCPVQAHEFVXVKT-UHFFFAOYSA-N 2-(2,4-difluorophenoxy)pyridin-3-amine Chemical compound NC1=CC=CN=C1OC1=CC=C(F)C=C1F LCPVQAHEFVXVKT-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 239000001116 FEMA 4028 Substances 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229920001774 Perfluoroether Polymers 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 description 1
- 235000011175 beta-cyclodextrine Nutrition 0.000 description 1
- 229960004853 betadex Drugs 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 239000007844 bleaching agent Substances 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000003517 fume Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000001744 histochemical effect Effects 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- RWPGFSMJFRPDDP-UHFFFAOYSA-L potassium metabisulfite Chemical compound [K+].[K+].[O-]S(=O)S([O-])(=O)=O RWPGFSMJFRPDDP-UHFFFAOYSA-L 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 230000005070 ripening Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 239000001509 sodium citrate Substances 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
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Substances [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
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
-
- 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
-
- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Detergent Compositions (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Electrotherapy Devices (AREA)
- Control And Other Processes For Unpacking Of Materials (AREA)
Description
ヘマトキシリン溶液(グリコール安定化溶液中、<1% ヘマトキシリン染料、<4% 硫酸アルミニウム、<0.1% ヨウ素酸ナトリウム、<1% ハイドロキノン、及び<2% β−シクロデキストリン水和物)の6ヶ月を経た未開封瓶を入手した。溶液を瓶から排出し、瓶を脱イオン水で3回濯いだ。ヘマトキシリン溶液が接触した瓶の内側には顕著な沈殿残渣が存在していた。瓶を切り開き、残留脱イオン水を蒸発させた。沈殿残渣を含んでいた瓶の部分を入手し、スポッティングテストに使用した。
パーフルオロアルコキシポリマー(PFA)チューブの12インチ片を、ヘマトキシリン水溶液[25% エチレングリコール(v/v)、20mM ヘマトキシリン、3.3mM ヨウ素酸ナトリウム、20mM 硫酸アルミニウム18水和物、85mM ハイドロキノン及び10mM β−シクロデキストリン60水和物、pH約2.5]で満たした。連結器でチューブの両端を連結し、ループを形成した。チューブ状ループを、約14日間にわたり60℃に保たれた炉内に置いた。この期間に、加熱によりヘマテインが沈殿して、チューブ状ループの内側を被覆する。この加熱インキュベーション期間の後、チューブ状ループを取り出し、排水し、脱イオン水で濯いだ。次にループを開放したままにして乾燥させた。これにより、チューブの内壁を被覆するヘマテイン沈殿物の暗色の被覆を伴ったチューブを得た。
3%過酸化水素溶液と0.5M炭酸ナトリウム溶液の1:1混合物を使用するスポットテスト実験は、優れた洗浄結果を示した。
0.125M塩化鉄(III)溶液、1Mリン酸緩衝液(pH=2)、0.64Mリン酸溶液、50%プロピレングリコール中0.1M塩酸、及び3%過酸化水素−50g/L炭酸ナトリウム溶液を比較する、チューブ状ループ実験を行った。3%過酸化水素−50g/L炭酸ナトリウム溶液はチューブを完璧に洗浄した。他の溶液ではすべて、チューブ壁に沈殿残渣が残った。
0.2Mシュウ酸溶液、0.2M亜硫酸ナトリウム溶液、0.2M二亜硫酸カリウム溶液、及び1M過硫酸ナトリウム−10g/L炭酸ナトリウム溶液を比較するスポットテスト実験を行った。1M過硫酸ナトリウム−10g/L炭酸ナトリウム溶液及び0.2Mシュウ酸溶液は優れた洗浄結果を示した。他の溶液は洗浄能力を示さなかった。
Claims (12)
- ヘマトキシリン自動染色システムからヘマテイン沈殿物を除去する方法であって、
過酸化物と水を含む液体酸化成分とアルカリ成分を提供し、ここで、液体酸化成分とアルカリ成分は分離されており;
容器内で液体酸化成分とアルカリ成分を合わせて、約8.9から10.5の間の範囲のpHを有するヘマテイン沈殿物除去液を形成し;
沈殿物除去液の容器を、ヘマトキシリン自動染色システムのヘマトキシリン分配要素がこれらの分配要素の内部表面を通って溶液を受け取りかつ移動させることを可能にする位置に置き;
自動染色システムを作動させて、沈殿物除去液を自動染色システムのヘマトキシリン分配要素の内部表面内に移動させ;
前記内部表面と少なくとも30分間接触させることを含む方法。 - 過酸化物が過酸化水素である、請求項1に記載の方法。
- 溶液の過酸化水素の重量百分率が1%から8%未満の範囲にある、請求項2に記載の方法。
- 溶液の過酸化水素の重量百分率が約3%から7%の間の範囲にある、請求項2に記載の方法。
- 溶液の過酸化水素の重量百分率が約6%である、請求項2に記載の方法。
- 溶液のpHが約9.5から10.5の間の範囲にある、請求項1から5のいずれか一項に記載の方法。
- 溶液のpHが約10である、請求項1から6のいずれか一項に記載の方法。
- アルカリ成分が炭酸ナトリウム、グリシンナトリウム及びリン酸三ナトリウムからなる群から選択されるアルカリ性剤を含む、請求項1から7のいずれか一項に記載の方法。
- アルカリ成分が液体形態であり、水をさらに含む、請求項8に記載の方法。
- 除去液容器が、洗浄されるヘマトキシリン自動染色システムのための染色試薬容器として構成される、請求項1から9のいずれか一項に記載の方法。
- 除去液容器が、ヘマトキシリン自動染色システムにおける試薬容器の置換により、ヘマトキシリン自動染色システムのヘマトキシリン分配要素がこれらの分配要素の内部表面を通って溶液を受け取りかつ移動させることを可能にするためのキーイング機能を備える、請求項10に記載の方法。
- 自動化された分配要素の内部表面から沈殿物除去液を除去し、自動染色システムをさらに作動することによって前記沈殿物除去液を染色試薬に置き換えることをさらに含む、請求項1から11のいずれか一項に記載の方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201461986386P | 2014-04-30 | 2014-04-30 | |
US61/986,386 | 2014-04-30 | ||
PCT/EP2015/059075 WO2015165850A1 (en) | 2014-04-30 | 2015-04-27 | Hematoxylin precipitate cleaning method and system |
Publications (2)
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JP2017519191A JP2017519191A (ja) | 2017-07-13 |
JP6263647B2 true JP6263647B2 (ja) | 2018-01-17 |
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JP2016564578A Active JP6263647B2 (ja) | 2014-04-30 | 2015-04-27 | ヘマトキシリン沈殿物の洗浄方法及びシステム |
Country Status (8)
Country | Link |
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US (1) | US10295443B2 (ja) |
EP (1) | EP3137215B1 (ja) |
JP (1) | JP6263647B2 (ja) |
CN (1) | CN106461517B (ja) |
AU (1) | AU2015252201B2 (ja) |
CA (1) | CA2941801C (ja) |
ES (1) | ES2918325T3 (ja) |
WO (1) | WO2015165850A1 (ja) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
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JP6731952B2 (ja) | 2015-05-22 | 2020-07-29 | ヴェンタナ メディカル システムズ, インク. | ヘマトキシリン溶液において生成される沈殿物を除去する又は沈殿物の形成を減少させるための方法及び装置 |
JP6583011B2 (ja) * | 2016-01-20 | 2019-10-02 | コニカミノルタ株式会社 | 酸性水溶液を用いた免疫染色スライドの洗浄方法 |
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