JP2008046127A - 試料の薄片から画像を取得する方法 - Google Patents
試料の薄片から画像を取得する方法 Download PDFInfo
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Images
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/225—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material using electron or ion
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- 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
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J2237/00—Discharge tubes exposing object to beam, e.g. for analysis treatment, etching, imaging
- H01J2237/30—Electron or ion beam tubes for processing objects
- H01J2237/317—Processing objects on a microscale
- H01J2237/3174—Etching microareas
- H01J2237/31745—Etching microareas for preparing specimen to be viewed in microscopes or analyzed in microanalysers
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Abstract
【解決手段】 試料(1)の表面層の画像を取得するステップと、前記試料の表面層を除去し、それにより次の薄片を表面にもたらすステップとが繰り返される、試料の薄片から画像を得る方法であって、少なくとも、前記試料の表面層の除去のうちの一つの除去の後に、前記試料は染色剤にさらされるステップを有することを特徴とする。
本方法は、とりわけ、走査電子顕微鏡カラム(20)と集束イオンビームカラム(10)とを備えた、粒子光学装置での使用に最適である。
試料は、その場所でOsO4のようなガスを前記試料に取り入れて、染色することができる。本方法は、第1染色剤に試料をさらして試料の第1画像を作り、第2の染色剤に追加的染色して、試料の画像を作ることにより、差異的な染色を形成することが可能となる。
【選択図】 図3
Description
・ 試料の表面層の画像を取得するステップ、及び
・ 試料の前記表面層を除去し、それにより次の薄片を表面にもたらすステップ、
が繰り返されることを含む、方法に関する。
John E. Scott and Constance R. "Dermatan sulphate-rich proteoglycan associates with rattendon collagen at the d band in the gap region" Orford, Biochem. J. 1981年、 p.213-216の ’materials and methods’の章 Y. Nagata 他 "Observation on backscattered electron image (BEI) of a scanning electron microscope (SEM) in semi-thin sections prepared for light microscopy" Tokai J. Exp. Clin. Med.、 1983年5月、p.167-174
2 試料ステージ
3 真空チャンバー
4 ガス注入システム
5 染色ユニット
10 イオン光学カラム
11 イオンの収束ビーム
20 電子光学カラム
21 電子の収束ビーム
30 二次電子検出器
31 X線検出器
32 反射電子検出器(BSD)
40 中央制御処理装置(CCPU)
50 観察スクリーン
Claims (12)
- 試料の薄片から画像を取得する方法であって、
前記試料の表面層の画像を取得するステップ、及び
前記試料の前記表面層を除去し、それにより次の薄片を表面にもたらすステップ、
が繰り返され、
表面層の除去のうちの少なくとも一つの除去の後に、前記試料が染色剤にさらされるステップ、
を有することを特徴とする方法。 - 前記画像は粒子光学装置で取得される、請求項1記載の方法。
- 前記表面層は、粒子ビームを用いて除去される、請求項1又は2記載の方法。
- 前記染色は、前記試料をガス又は蒸気にさらすことにより行われる、請求項1乃至3のいずれか一項記載の方法。
- 前記染色は、真空チャンバー内で又は少なくとも低圧のチャンバー内で、行われる、請求項4記載の方法。
- 前記試料は、粒子光学装置の一部分である真空チャンバー内で染色される、請求項5記載の方法。
- 前記試料は、二つ以上の染色剤にさらされる、請求項1乃至6のいずれか一項記載の方法。
- 前記試料の画像が、前記試料が一つのみの染色剤で染色されたときに取得され、また前記試料が追加の染色剤で染色されたときにも取得され、それにより差異的染色を可能にする、請求項7記載の方法。
- 前記試料の画像が、前記試料が一つの染色剤で染色されたときに、取得され、その後、前記表面層が除去されて、前記試料は他の染色剤にさらされ、そして、次に他の画像が取得され、それにより、一度に一つの染色剤で染色された前記試料の画像群を得る、請求項7記載の方法。
- 前記試料の前記表面層の画像が、染色された状態だけでなく染色されない状態でも、取得され、そして、そのようにして取得された画像は、染色された画像中の染色されない試料の地形学的及びトポロジー的な情報を除去するか、少なくとも、大きく減じるように、組み合わされる、請求項1乃至9のいずれか一項記載の方法。
- 前記染色剤は、重金属と有機基を有する高分子を含む、請求項1乃至10のいずれか一項記載の方法。
- ガス注入システムであって、粒子光学装置の真空チャンバー内に染色ガス又は蒸気を取り入れるために備えられた前記ガス注入システム。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP06118983A EP1890136A1 (en) | 2006-08-16 | 2006-08-16 | Method for obtaining images from slices of a specimen |
EP06118983.3 | 2006-08-16 |
Publications (2)
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JP2008046127A true JP2008046127A (ja) | 2008-02-28 |
JP5090102B2 JP5090102B2 (ja) | 2012-12-05 |
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JP2007211771A Active JP5090102B2 (ja) | 2006-08-16 | 2007-08-15 | 試料の薄片から画像を取得する方法 |
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Country | Link |
---|---|
US (2) | US7977631B2 (ja) |
EP (1) | EP1890136A1 (ja) |
JP (1) | JP5090102B2 (ja) |
CN (1) | CN101126726B (ja) |
DE (1) | DE602007011856D1 (ja) |
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JP2014160068A (ja) * | 2013-02-19 | 2014-09-04 | Fei Co | 蛍光マーカの原位置再活性化 |
JP2018194469A (ja) * | 2017-05-18 | 2018-12-06 | 住友ゴム工業株式会社 | 試料の観察方法 |
KR20190060101A (ko) * | 2017-11-24 | 2019-06-03 | 주식회사 엘지화학 | Ssbr 내 sbr 상과 br 상의 tem 분석방법 |
KR102401912B1 (ko) | 2017-11-24 | 2022-05-24 | 주식회사 엘지화학 | Ssbr 내 sbr 상과 br 상의 tem 분석방법 |
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US8431896B2 (en) | 2013-04-30 |
EP1890136A1 (en) | 2008-02-20 |
CN101126726B (zh) | 2012-11-14 |
US7977631B2 (en) | 2011-07-12 |
CN101126726A (zh) | 2008-02-20 |
JP5090102B2 (ja) | 2012-12-05 |
US20110226819A1 (en) | 2011-09-22 |
DE602007011856D1 (de) | 2011-02-24 |
US20080088831A1 (en) | 2008-04-17 |
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