JP2014021109A - 高圧冷凍材料からの電子顕微鏡用試料の作製 - Google Patents
高圧冷凍材料からの電子顕微鏡用試料の作製 Download PDFInfo
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- JP2014021109A JP2014021109A JP2013143193A JP2013143193A JP2014021109A JP 2014021109 A JP2014021109 A JP 2014021109A JP 2013143193 A JP2013143193 A JP 2013143193A JP 2013143193 A JP2013143193 A JP 2013143193A JP 2014021109 A JP2014021109 A JP 2014021109A
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- 239000000463 material Substances 0.000 title claims abstract description 24
- 239000000523 sample Substances 0.000 claims abstract description 71
- 238000000034 method Methods 0.000 claims abstract description 33
- 238000001816 cooling Methods 0.000 claims abstract description 10
- 238000005520 cutting process Methods 0.000 claims abstract description 10
- 238000010792 warming Methods 0.000 claims abstract description 10
- 238000010884 ion-beam technique Methods 0.000 claims abstract description 9
- 230000009477 glass transition Effects 0.000 claims abstract description 5
- 239000007788 liquid Substances 0.000 claims description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 claims description 8
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims description 7
- 229910052802 copper Inorganic materials 0.000 claims description 7
- 239000010949 copper Substances 0.000 claims description 7
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 4
- 229910052751 metal Inorganic materials 0.000 claims description 4
- 239000002184 metal Substances 0.000 claims description 4
- 239000001294 propane Substances 0.000 claims description 4
- OTMSDBZUPAUEDD-UHFFFAOYSA-N Ethane Chemical compound CC OTMSDBZUPAUEDD-UHFFFAOYSA-N 0.000 claims description 3
- 229910052782 aluminium Inorganic materials 0.000 claims description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 2
- 239000012520 frozen sample Substances 0.000 claims description 2
- 238000007654 immersion Methods 0.000 claims description 2
- 238000007689 inspection Methods 0.000 claims description 2
- 229910052742 iron Inorganic materials 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims 1
- 238000003801 milling Methods 0.000 abstract description 6
- 238000007710 freezing Methods 0.000 description 7
- 230000008014 freezing Effects 0.000 description 7
- 239000012620 biological material Substances 0.000 description 5
- 230000005540 biological transmission Effects 0.000 description 5
- 238000001125 extrusion Methods 0.000 description 5
- 238000005057 refrigeration Methods 0.000 description 3
- RTAQQCXQSZGOHL-UHFFFAOYSA-N Titanium Chemical compound [Ti] RTAQQCXQSZGOHL-UHFFFAOYSA-N 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 229930195733 hydrocarbon Natural products 0.000 description 2
- 150000002430 hydrocarbons Chemical class 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 238000000604 cryogenic transmission electron microscopy Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010894 electron beam technology Methods 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- CKHJYUSOUQDYEN-UHFFFAOYSA-N gallium(3+) Chemical compound [Ga+3] CKHJYUSOUQDYEN-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229910000489 osmium tetroxide Inorganic materials 0.000 description 1
- 239000012285 osmium tetroxide Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 125000005289 uranyl group Chemical group 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
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B23—MACHINE TOOLS; METAL-WORKING NOT OTHERWISE PROVIDED FOR
- B23K—SOLDERING OR UNSOLDERING; WELDING; CLADDING OR PLATING BY SOLDERING OR WELDING; CUTTING BY APPLYING HEAT LOCALLY, e.g. FLAME CUTTING; WORKING BY LASER BEAM
- B23K15/00—Electron-beam welding or cutting
- B23K15/08—Removing material, e.g. by cutting, by hole drilling
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
- G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
- G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Mechanical Engineering (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
【解決手段】ガラス転移温度Tg未満の温度T1でガラス化した試料材料を高圧キャピラリに供する段階102、前記キャピラリを切断する段階104、前記キャピラリを、温度T1以上温度Tg以下の温度T2にまで温める段階、前記キャピラリを温度T2未満の温度T3にまで冷却して、前記材料を前記キャピラリから押し出す段階、及び、温度Tg未満の温度で前記の押し出された試料材料から試料を解放する段階を有する。このようにして押し出された材料から、試料が、たとえばイオンビームミリングによって薄片となるように切断され得る。
【選択図】図1
Description
− ガラス転移温度Tg未満の温度T1でガラス化した試料材料を高圧キャピラリに供する段階、及び、
− 前記キャピラリを切断する段階、
を有する。
電子の密な材料による)電子顕微鏡の着色/標識付け段階が導入されても良い。
204 押し出された中心部
206 押し出された分の長さ
Claims (9)
- 高圧冷凍試料材料を有するキャピラリから試料を作製する方法であって:
ガラス転移温度Tg未満の温度T1でガラス化した試料材料を高圧キャピラリに供する段階;
前記キャピラリを切断する段階;
前記キャピラリを温度T1以上温度Tg以下の温度T2にまで温める段階;
前記キャピラリを温度T2未満の温度T3にまで冷却して、前記材料を前記キャピラリから押し出す段階;及び、
温度Tg未満の温度で前記の押し出された試料材料から試料を解放する段階;
を有することを特徴とする方法。 - 前記キャピラリを温度T2よりも低い温度T3に冷却する段階は、前記キャピラリを温度T2よりも低い温度の液体−具体的には液体エタン、液体プロパン、又は液体窒素−に浸漬させる段階を有する、請求項1に記載の方法。
- 前記キャピラリが金属キャピラリである、請求項1又は2に記載の方法。
- 前記金属が、鉄、銅、及び/又はアルミニウムを含む、請求項3に記載の方法。
- 前記温度T2と温度T3との差異は20Kよりも大きく、50Kよりも大きいことが好ましい、請求項1乃至4のうちいずれか一項に記載の方法。
- 前記試料を解放する段階は、集束イオンビームにより切断する段階を有する、請求項1乃至5のうちいずれか一項に記載の方法。
- 前記の解放された試料は薄片である、請求項1乃至6のうちいずれか一項に記載の方法。
- 前記電子顕微鏡内での検査段階をさらに有する、請求項7に記載の方法。
- 前記キャピラリを温度T1以上温度Tg以下の温度T2にまで温める段階と、該温める段階に続く前記キャピラリを温度T2未満の温度T3にまで冷却して、前記材料を前記キャピラリから押し出す段階の後、さらに前記キャピラリを温度T1以上温度Tg以下の温度T2にまで温める段階と、該温める段階に続いてさらに前記キャピラリを温度T2未満の温度T3にまで冷却して、前記材料を前記キャピラリから押し出す段階が実行される、請求項1乃至8のうちいずれか一項に記載の方法。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP12176254.6A EP2685233A1 (en) | 2012-07-13 | 2012-07-13 | Forming an electron microscope sample from high-pressure frozen material |
EP12176254.6 | 2012-07-13 |
Publications (2)
Publication Number | Publication Date |
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JP2014021109A true JP2014021109A (ja) | 2014-02-03 |
JP6294017B2 JP6294017B2 (ja) | 2018-03-14 |
Family
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2013143193A Active JP6294017B2 (ja) | 2012-07-13 | 2013-07-09 | 高圧冷凍材料からの電子顕微鏡用試料の作製 |
Country Status (4)
Country | Link |
---|---|
US (1) | US8674323B2 (ja) |
EP (2) | EP2685233A1 (ja) |
JP (1) | JP6294017B2 (ja) |
CN (1) | CN103543045B (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2017053694A (ja) * | 2015-09-09 | 2017-03-16 | 日本電子株式会社 | 加工方法 |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2813835B1 (en) | 2013-06-14 | 2016-09-07 | Fei Company | Method of welding a frozen aqueous sample to a microprobe |
RU2663732C1 (ru) * | 2017-05-04 | 2018-08-09 | Игорь Михайлович Рулев | Способ автоматического отбора и упаковки микробиологических объектов |
CN110006934A (zh) | 2017-12-28 | 2019-07-12 | Fei 公司 | 通过等离子体聚焦离子束处理生物低温样品的方法、装置和系统 |
CN110823931B (zh) * | 2019-10-16 | 2020-10-27 | 中国科学院生物物理研究所 | 一种冷冻电镜样品制备方法 |
DE202023100215U1 (de) | 2023-01-17 | 2023-02-06 | Cbg Composites Gmbh | Faserbetonprodukt auf Basis von Basaltfasern mit plastifizierender Wirkung |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10221228A (ja) * | 1997-01-13 | 1998-08-21 | Leica Ag | 含水試料の試料ホルダおよびその使用法 |
JP2009014719A (ja) * | 2007-06-29 | 2009-01-22 | Fei Co | マニピュレータへのサンプル取付け方法 |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5582944A (en) * | 1978-12-19 | 1980-06-23 | Toukiyouto | Preparation of replica thin-film for specimen of electron microscope |
US6881363B2 (en) * | 2001-12-07 | 2005-04-19 | Symyx Technologies, Inc. | High throughput preparation and analysis of materials |
EP1863066A1 (en) | 2006-05-29 | 2007-12-05 | FEI Company | Sample carrier and sample holder |
EP2009420A1 (en) * | 2007-06-29 | 2008-12-31 | FEI Company | Method for attaching a sample to a manipulator |
EP2009421A1 (en) * | 2007-06-29 | 2008-12-31 | FEI Company | Method for separating a lamella for TEM inspection from a core sample |
CN101648085B (zh) * | 2008-08-14 | 2012-03-21 | 浙江工业大学 | 毛细管反应器的制作设备及制作方法 |
EP2402475A1 (en) * | 2010-06-30 | 2012-01-04 | Fei Company | Beam-induced deposition at cryogenic temperatures |
US8884248B2 (en) * | 2012-02-13 | 2014-11-11 | Fei Company | Forming a vitrified sample for electron microscopy |
-
2012
- 2012-07-13 EP EP12176254.6A patent/EP2685233A1/en not_active Withdrawn
-
2013
- 2013-07-03 US US13/934,925 patent/US8674323B2/en active Active
- 2013-07-09 JP JP2013143193A patent/JP6294017B2/ja active Active
- 2013-07-12 CN CN201310301331.4A patent/CN103543045B/zh active Active
- 2013-07-12 EP EP13176233.8A patent/EP2685234B1/en active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH10221228A (ja) * | 1997-01-13 | 1998-08-21 | Leica Ag | 含水試料の試料ホルダおよびその使用法 |
JP2009014719A (ja) * | 2007-06-29 | 2009-01-22 | Fei Co | マニピュレータへのサンプル取付け方法 |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2017053694A (ja) * | 2015-09-09 | 2017-03-16 | 日本電子株式会社 | 加工方法 |
Also Published As
Publication number | Publication date |
---|---|
US20140014834A1 (en) | 2014-01-16 |
EP2685233A1 (en) | 2014-01-15 |
EP2685234B1 (en) | 2014-12-17 |
JP6294017B2 (ja) | 2018-03-14 |
CN103543045B (zh) | 2017-02-08 |
CN103543045A (zh) | 2014-01-29 |
US8674323B2 (en) | 2014-03-18 |
EP2685234A1 (en) | 2014-01-15 |
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