JPH05503149A - 2光子レーザ走査顕微鏡 - Google Patents
2光子レーザ走査顕微鏡Info
- Publication number
- JPH05503149A JPH05503149A JP3500520A JP50052091A JPH05503149A JP H05503149 A JPH05503149 A JP H05503149A JP 3500520 A JP3500520 A JP 3500520A JP 50052091 A JP50052091 A JP 50052091A JP H05503149 A JPH05503149 A JP H05503149A
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- light
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- fluorescence
- microscope
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01—MEASURING; TESTING
- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
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- G—PHYSICS
- G11—INFORMATION STORAGE
- G11B—INFORMATION STORAGE BASED ON RELATIVE MOVEMENT BETWEEN RECORD CARRIER AND TRANSDUCER
- G11B7/00—Recording or reproducing by optical means, e.g. recording using a thermal beam of optical radiation by modifying optical properties or the physical structure, reproducing using an optical beam at lower power by sensing optical properties; Record carriers therefor
- G11B7/004—Recording, reproducing or erasing methods; Read, write or erase circuits therefor
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- G—PHYSICS
- G11—INFORMATION STORAGE
- G11B—INFORMATION STORAGE BASED ON RELATIVE MOVEMENT BETWEEN RECORD CARRIER AND TRANSDUCER
- G11B7/00—Recording or reproducing by optical means, e.g. recording using a thermal beam of optical radiation by modifying optical properties or the physical structure, reproducing using an optical beam at lower power by sensing optical properties; Record carriers therefor
- G11B7/12—Heads, e.g. forming of the optical beam spot or modulation of the optical beam
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- G—PHYSICS
- G11—INFORMATION STORAGE
- G11B—INFORMATION STORAGE BASED ON RELATIVE MOVEMENT BETWEEN RECORD CARRIER AND TRANSDUCER
- G11B7/00—Recording or reproducing by optical means, e.g. recording using a thermal beam of optical radiation by modifying optical properties or the physical structure, reproducing using an optical beam at lower power by sensing optical properties; Record carriers therefor
- G11B7/12—Heads, e.g. forming of the optical beam spot or modulation of the optical beam
- G11B7/135—Means for guiding the beam from the source to the record carrier or from the record carrier to the detector
- G11B7/1362—Mirrors
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N21/6408—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence
- G01N2021/6415—Fluorescence; Phosphorescence with measurement of decay time, time resolved fluorescence with two excitations, e.g. strong pump/probe flash
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N21/631—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited using photolysis and investigating photolysed fragments
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Optics & Photonics (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microscoopes, Condenser (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Lasers (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
Description
Claims (18)
- 1.以下のものを含むレーザ走査顕微鏡画像化されるべき目的物を受ける対象物 面;上記目的物は該目的物の蛍光特性を生成するため短波長スペクトラル領域の 光による励起に応答する蛍光手段を含んでいる、上記目標面に隣接して位置する レンズ手段;長波長スペクトラル領域のサブピコ秒の単色コヒーレント光パルス 源; 検出手段; 上記コヒーレント光を上記レンズ手段を含む光路に沿って上記目的物に入射させ るべく指向させるミラー手段;上記長波長の光パルスは上記目的物内で蛍光を発 生させるに十分な瞬間的パワーを与え、該蛍光は上記光路上を伝播して上記検出 手段に至る出力光を与える。
- 2.請求項1の顕微鏡において、上記光源は高い繰返率で高い瞬間パワーのパル スを発生し、上記目的物は上記短波長の入射光によって供給されたエネルギーに 競り合って、少なくとも2つの長波長光入射パルスからエネルギーを吸収するも の。
- 3.請求項2の顕微鏡において、上記光源からの長波長光は上記レンズ手段によ り上記目的物内にサブミクロンの直径にまで結焦され、上記目的物内に蛍光を発 生させるのに十分な高強度を生成するもの。
- 4.請求項2の顕微鏡において、上記レンズ手段は上記目標面の対向面に収束し 発散する光を発生すべく、上記長波長光を円錐形状に結焦し、それによって上記 長波長光は上記目標面上の焦点に結焦されるもの。
- 5.請求項2の顕微鏡において、上記目的物は紫外線波長スペクトラル領域に単 一の光子吸収ピークを有する蛍光発光団であり、赤色波長スペクトラル領域にお いて2個の光子を吸収することができるもの。
- 6.請求項5の顕微鏡において、上記レンズ手段は、上記目的物内の焦点に上記 長波長光を結焦し、該焦点の周囲の限定された楕円体内に蛍光を励起する光強度 を発生するもの。
- 7.以下のものを含むレーザ走査顕微鏡、所定の波長の光による単一光子励起に 対応する吸収ピークを有する目的物を受ける目標面; 上記目標面に隣接して位置するレンズ手段;長波長のサブピコ秒の光パルスのレ ーザ光源;該光は上記所定の波長の約2倍の波長を有する、上記光パルスを上記 目的物に入射させるべく、上記レンズ手段を含む光路に沿って上記光パルスを指 向させるミラー手段;上記レンズ手段は上記目的物内の焦点に上記光パルスを結 焦し、上記光パルスの強度は上記単一光子吸収ピークに対応する単一光子励起エ ネルギーレベルに等価な2光子励起エネルギーレベルを上記焦点領域に生成する 。
- 8.請求項7の顕微鏡において、上記目的物は上記吸収ピークを有する蛍光発光 団を含むもの。
- 9.請求項8の顕微鏡において、上記蛍光発光団は蛍光を発生する所定の強度を 与える入射光子に応答するもの。
- 10.請求項9の顕微鏡において、上記入射光パルスは光エネルギー有する光子 を上記目的物に与えるとともに、上記入射光の2光子の結合されたエネルギーが 蛍光を発生させるために必要とされるもの。
- 11.請求項10の顕微鏡において、上記レンズ手段は目的物内において異なる 深さの焦点を選択しうるようになっているもの。
- 12.請求項11の顕微鏡において、目的物に関し、上記焦点を移動させるため の走査手段をさらに含むもの。
- 13.請求項12の顕微鏡において、上記目的物の蛍光を検出するための光路内 の光に感応する検出手段をさらに含むもの。
- 14.請求項13の顕微鏡において、上記検出手段は、上記蛍光に感応する光感 知アレイであるもの。
- 15.請求項7の顕微鏡において、上記目的物は上記光パルスによって上記焦点 に生成される2光子励起エネルギーレベルに反応する生物細胞であるもの。
- 16.請求項15の顕微鏡において、上記目的物は生物学的に活性な化学物質の 局所的な放出を発生するために上記焦点における上記光パルスに反応するもの。
- 17.請求項7の顕微鏡において、上記目的物は光子により活性化されうる試薬 であるもの。
- 18.請求項7の顕微鏡において、上記目的物は単一光源からの2光子エネルギ ーに応答する光メモリであるもの。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US436,045 | 1989-11-14 | ||
US07/436,045 US5034613A (en) | 1989-11-14 | 1989-11-14 | Two-photon laser microscopy |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH05503149A true JPH05503149A (ja) | 1993-05-27 |
JP2848952B2 JP2848952B2 (ja) | 1999-01-20 |
Family
ID=23730872
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP3500520A Expired - Fee Related JP2848952B2 (ja) | 1989-11-14 | 1990-11-13 | 2光子レーザ走査顕微鏡 |
Country Status (9)
Country | Link |
---|---|
US (1) | US5034613A (ja) |
EP (2) | EP0500717B2 (ja) |
JP (1) | JP2848952B2 (ja) |
AT (2) | ATE253727T1 (ja) |
DE (2) | DE69034117T2 (ja) |
DK (1) | DK0807814T3 (ja) |
ES (2) | ES2123501T5 (ja) |
HK (1) | HK1010411A1 (ja) |
WO (1) | WO1991007651A1 (ja) |
Cited By (12)
Publication number | Priority date | Publication date | Assignee | Title |
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JPH0915171A (ja) * | 1995-03-16 | 1997-01-17 | Bio Rad Lab Inc | 蛍光結像システム及び蛍光結像方法 |
JPH10206742A (ja) * | 1996-11-21 | 1998-08-07 | Olympus Optical Co Ltd | レーザ走査顕微鏡 |
JP2000193889A (ja) * | 1998-12-28 | 2000-07-14 | Univ Osaka | 多光子顕微鏡 |
JP2002113786A (ja) * | 2000-06-05 | 2002-04-16 | Leica Microsystems Heidelberg Gmbh | 三次元物体の生成方法 |
US6943944B2 (en) | 2000-09-29 | 2005-09-13 | Olympus Optical Co., Ltd. | Laser microscope and laser pulse width control method |
JP2007086047A (ja) * | 2005-08-22 | 2007-04-05 | National Institute Of Advanced Industrial & Technology | 分光化学分析及び3次元形状観察法並びにその装置 |
JP2007510176A (ja) * | 2003-10-30 | 2007-04-19 | カール ツァイス イエナ ゲゼルシャフト ミット ベシュレンクテル ハフツング | ノン・デスキャンド検出光路および/または観察光路を備えるレーザ走査型顕微鏡 |
JP2007538289A (ja) * | 2004-05-19 | 2007-12-27 | ゲーリー、ブルッカー | 共焦点励起平面を有した広視野多光子顕微鏡のための方法およびシステム |
WO2009016806A1 (ja) * | 2007-07-27 | 2009-02-05 | Nikon Corporation | マルチフォトンレーザ走査顕微鏡装置 |
JP2010175464A (ja) * | 2009-01-30 | 2010-08-12 | Toshiba Corp | 顕微鏡および観測方法 |
WO2011162187A1 (ja) * | 2010-06-23 | 2011-12-29 | 浜松ホトニクス株式会社 | 画像生成装置 |
JP2015148442A (ja) * | 2014-02-04 | 2015-08-20 | 株式会社Ihi | 物質特定方法および物質特定システム |
Families Citing this family (343)
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JP4621893B2 (ja) * | 2005-08-22 | 2011-01-26 | 独立行政法人産業技術総合研究所 | 物体の調査方法及び調査装置 |
WO2009016806A1 (ja) * | 2007-07-27 | 2009-02-05 | Nikon Corporation | マルチフォトンレーザ走査顕微鏡装置 |
JP2010175464A (ja) * | 2009-01-30 | 2010-08-12 | Toshiba Corp | 顕微鏡および観測方法 |
WO2011162187A1 (ja) * | 2010-06-23 | 2011-12-29 | 浜松ホトニクス株式会社 | 画像生成装置 |
JP2012008261A (ja) * | 2010-06-23 | 2012-01-12 | Hamamatsu Photonics Kk | 画像生成装置 |
JP2015148442A (ja) * | 2014-02-04 | 2015-08-20 | 株式会社Ihi | 物質特定方法および物質特定システム |
Also Published As
Publication number | Publication date |
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ES2123501T5 (es) | 2004-07-16 |
EP0807814A1 (en) | 1997-11-19 |
DE69032621T2 (de) | 1999-03-25 |
US5034613A (en) | 1991-07-23 |
WO1991007651A1 (en) | 1991-05-30 |
DE69032621D1 (de) | 1998-10-08 |
ES2210414T3 (es) | 2004-07-01 |
EP0500717A1 (en) | 1992-09-02 |
EP0500717B1 (en) | 1998-09-02 |
DE69034117T2 (de) | 2004-08-19 |
EP0500717B2 (en) | 2003-11-19 |
ATE170636T1 (de) | 1998-09-15 |
ATE253727T1 (de) | 2003-11-15 |
DE69034117D1 (de) | 2003-12-11 |
DK0807814T3 (da) | 2004-03-15 |
ES2123501T3 (es) | 1999-01-16 |
JP2848952B2 (ja) | 1999-01-20 |
DE69032621T3 (de) | 2004-08-19 |
EP0500717A4 (en) | 1992-12-09 |
HK1010411A1 (en) | 1999-06-17 |
EP0807814B1 (en) | 2003-11-05 |
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