JP2016503892A - 発光団を有する試料構造の高空間分解イメージング方法 - Google Patents
発光団を有する試料構造の高空間分解イメージング方法 Download PDFInfo
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- 238000003384 imaging method Methods 0.000 title claims abstract description 14
- 238000004020 luminiscence type Methods 0.000 claims abstract description 107
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- 230000002123 temporal effect Effects 0.000 claims description 2
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- 125000003636 chemical group Chemical group 0.000 description 1
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G—PHYSICS
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
- G01N2021/6419—Excitation at two or more wavelengths
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
- G01N2021/6421—Measuring at two or more wavelengths
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- G—PHYSICS
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Abstract
Description
Claims (15)
- 発光団(1)を有する試料(3)の構造(2)の高空間分解イメージング方法であり、
‐励起可能な電子基底状態から励起発光状態に前記発光団(1)を励起するルミネッセンス励起光(7)を測定範囲(5)の前記試料(3)に当て、
‐前記励起発光状態から前記励起可能な電子基底状態に前記発光団(1)を戻すルミネッセンス減衰光(8)の強度分布であり、局所的最小値(9)を有する強度分布を前記測定範囲(5)の前記試料(3)に当て、
‐前記測定範囲(5)から放出された発光光(10)を記録し、記録された前記発光光(10)を前記試料(3)の前記局所的最小値(9)の位置に割り当てる方法であって、
前記測定範囲(5)の前記試料(3)に、前記ルミネッセンス励起光を当てる前に、前記励起可能な電子基底状態から保護状態に前記発光団(1)を変換する励起阻止光(4)の強度分布を当て、前記保護状態では、前記発光団(1)が前記ルミネッセンス励起光(7)と前記ルミネッセンス減衰光(8)とによって電子励起から保護されており、前記励起阻止光(4)の前記強度分布は、前記ルミネッセンス減衰光(8)の前記強度分布の前記局所的最小値と重なる局所的最小値(6)を有していることを特徴とする、方法。 - 前記ルミネッセンス励起光(7)、前記ルミネッセンス減衰光(8)及び前記励起阻止光(4)が、異なる波長を有していることを特徴とする、請求項1に記載の方法。
- 前記ルミネッセンス減衰光(8)及び前記励起阻止光(4)が、同じ波長を有していることを特徴とする、請求項1に記載の方法。
- 前記ルミネッセンス減衰光(8)及び前記励起阻止光(4)が、同時に前記試料(3)に当てられることを特徴とする、請求項3に記載の方法。
- 前記ルミネッセンス励起光(7)が、パルスで前記試料(3)に当てられること、及び前記励起阻止光(4)と前記ルミネッセンス減衰光(8)とが、互いに独立して、パルスで又は連続して前記試料(3)に当てられることを特徴とする、請求項1〜4のうちいずれか一項に記載の方法。
- 前記発光光(10)が、前記ルミネッセンス励起光(7)の各パルスの後で時間分解能によって記録されることを特徴とする、請求項5に記載の方法。
- 前記励起阻止光(4)が、立体配座変化を利用して、前記発光団(1)を前記励起可能な電子基底状態から前記保護状態へと変換することを特徴とする、請求項1〜6のうちいずれか一項に記載の方法。
- 前記発光団(1)が、前記励起阻止光(4)によってオフに切り替えられる発光団(1)であることを特徴とする、請求項1〜7のうちいずれか一項に記載の方法。
- 前記発光団(1)が、切替え可能な蛍光タンパク質であることを特徴とする、請求項1〜8のうちいずれか一項に記載の方法。
- 前記測定範囲(5)の前記試料(3)に前記励起阻止光(4)を当てる前に、前記励起可能光の強度分布を当て、前記励起可能光は、少なくとも前記ルミネッセンス減衰光(8)の前記強度分布の前記局所的最小値(9)の部分で、前記発光団(1)を前記励起可能な電子基底状態に変換することを特徴とする、請求項7〜9のうちいずれか一項に記載の方法。
- 前記発光団(1)を前記保護状態に変換するため、前記保護状態にある前記発光団(1)が前記ルミネッセンス励起光(7)について少なくとも1つの係数(2)の分だけ低下した吸収断面積を有するように、前記励起阻止光(4)が前記発光団(1)の前記励起可能な電子基底状態を妨げることを特徴とする、請求項1〜6のうちいずれか一項に記載の方法。
- 前記発光団(1)の前記電子基底状態が、運動パルス及び/又は振動の伝達によって妨げられていることを特徴とする、請求項11に記載の方法。
- 前記運動パルス及び/又は振動が、前記励起阻止光(4)によって励起されたモジュレータから発生することを特徴とする、請求項12に記載の方法。
- 前記妨げられている電子基底状態では、前記発光団(1)の原子の配列が障害を受けていることを特徴とする、請求項11〜13のうちいずれか一項に記載の方法。
- 前記障害を受けている電子基底状態の前記発光団(1)が、熱平衡にはないことを特徴とする、請求項11〜14のうちいずれか一項に記載の方法。
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DE102013100172.6A DE102013100172A1 (de) | 2013-01-09 | 2013-01-09 | Verfahren zum räumlich hochaufgelösten Abbilden einer einen Luminophor aufweisenden Struktur einer Probe |
DE102013100172.6 | 2013-01-09 | ||
PCT/EP2014/050272 WO2014108455A1 (de) | 2013-01-09 | 2014-01-09 | Verfahren zum räumlich hochaufgelösten abbilden einer einen luminophor aufweisenden struktur einer probe |
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US (1) | US9267888B2 (ja) |
EP (1) | EP2943776B1 (ja) |
JP (1) | JP6281775B2 (ja) |
CN (1) | CN104919303B (ja) |
DE (1) | DE102013100172A1 (ja) |
HK (1) | HK1214350A1 (ja) |
WO (1) | WO2014108455A1 (ja) |
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JP2019507921A (ja) * | 2016-03-07 | 2019-03-22 | マックス−プランク−ゲゼルシャフト ツア フェルデルング デア ヴィッセンシャフテン イー.ヴイ.Max−Planck−Gesellschaft zur Forderung der Wissenschaften e.V. | 多数の対象の対象固有の場所間の輸送方法 |
JP7027316B2 (ja) | 2016-03-07 | 2022-03-01 | マックス-プランク-ゲゼルシャフト ツア フェルデルング デア ヴィッセンシャフテン イー.ヴイ. | 多数の対象の対象固有の場所間の輸送方法 |
JP2018169387A (ja) * | 2017-03-07 | 2018-11-01 | マックス−プランク−ゲゼルシャフト ツア フェルデルング デア ヴィッセンシャフテン イー.ヴイ.Max−Planck−Gesellschaft zur Forderung der Wissenschaften e.V. | ナノスケール構造体を空間的に測定するための方法および装置 |
JP7201327B2 (ja) | 2017-03-07 | 2023-01-10 | マックス-プランク-ゲゼルシヤフト・ツーア・フェルデルング・デア・ヴィッセンシャフテン・アインゲトラーゲナー・フェライン | ナノスケール構造体の空間的測定方法及び装置 |
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US20150308955A1 (en) | 2015-10-29 |
JP6281775B2 (ja) | 2018-02-21 |
CN104919303A (zh) | 2015-09-16 |
EP2943776B1 (de) | 2019-05-22 |
HK1214350A1 (zh) | 2016-07-22 |
DE102013100172A1 (de) | 2014-07-10 |
US9267888B2 (en) | 2016-02-23 |
EP2943776A1 (de) | 2015-11-18 |
WO2014108455A1 (de) | 2014-07-17 |
CN104919303B (zh) | 2017-10-31 |
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