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3-dimensional confocal microscopy apparatus and focal plane scanning and aberration correction unit

外国特許コード F130007601
整理番号 AF02-09WO
掲載日 2013年8月9日
出願国 欧州特許庁(EPO)
出願番号 11824911
公報番号 2618197
公報番号 2618197
出願日 平成23年7月27日(2011.7.27)
公報発行日 平成25年7月24日(2013.7.24)
公報発行日 平成28年6月8日(2016.6.8)
国際出願番号 JP2011067719
国際公開番号 WO2012035903
国際出願日 平成23年7月27日(2011.7.27)
国際公開日 平成24年3月22日(2012.3.22)
優先権データ
  • 特願2010-208971 (2010.9.17) JP
  • 2011JP067719 (2011.7.27) WO
発明の名称 (英語) 3-dimensional confocal microscopy apparatus and focal plane scanning and aberration correction unit
発明の概要(英語) Provided is a 3-dimensional confocal microscopy apparatus which is manufactured by combining a confocal microscope and an optical tweezers technique, wherein a pair of lenses for focal plane displacement where one lens is movable in the optical axis direction is arranged between a fixed objective lens and a fluorescent light imaging camera, and the 3-dimensional confocal microscopy apparatus also includes a mean which corrects the aberration of a fluorescent confocal image obtained by the fluorescent imaging camera.
Accordingly, it is possible to provide a 3-dimensional confocal microscopy apparatus which can acquire a 3-dimensional image of a specimen during a manipulation of the specimen using optical tweezers without affecting an optical trap.
特許請求の範囲(英語) [claim1]
1. A 3-dimensional confocal microscopy apparatus comprising: a light source for a first laser (1, 10) for confocal imaging excitation; a 2-dimensional scanning optical system for scanning the first laser on a specimen; an objective lens (2, 12) for focusing the first laser supplied from the light source on a fluorescent specimen which is an object to be observed; a fluorescent light imaging camera (16) for detecting a fluorescent confocal microscopy image based on a fluorescent light emitted from the fluorescent specimen; a pair of lenses (14, 15); lens drive means (17); a beam splitter or a dichroic mirror (20) and a light source (18) for a second laser; wherein
the pair of lenses (14, 15) for focal plane displacement is arranged between the objective lens and the fluorescent light imaging camera, the pair of lenses for focal plane displacement being constituted of an intermediate imaging lens (14) which forms an intermediate image of the fluorescent confocal microscopy image of the specimen and an intermediate objective lens (15) arranged such that a focal point of the intermediate objective lens overlaps with a focal point of the intermediate imaging lens on the same optical axis, and at least one lens out of these lenses being movable in an optical axis direction by the lens drive means (17),
the 3-dimensional confocal microscopy apparatus further includes a computer having an arithmetic operation means for acquiring a 3-dimensional structural image of the specimen by correcting aberration of the fluorescent confocal microscopy image caused by the movement of the lens of the pair of lenses for focal plane displacement in real time using the fluorescent confocal microscopy image acquired by the fluorescent light imaging camera and aberration correction data which is acquired in advance by performing provisional measurement of a sample specimen, and a means which displays a corrected image obtained by the arithmetic operation means, wherein
the beam splitter or a dichroic mirror (20) is provided between the pair of lenses for focal plane displacement and the objective lens, and the second laser supplied from the light source (18) for a second laser for optical tweezers passes through the objective lens via the beam splitter or the dichroic mirror and is focused on the specimen.
[claim2]
2. The 3-dimensional confocal microscopy apparatus according to claim 1, wherein the 2-dimensional scanning optical system includes a nipkow disc or a line scanning type scanning system.
[claim3]
3. The 3-dimensional confocal microscopy apparatus according to claim 1 or 2, wherein the lens drive means for the pair of lenses for focal plane displacement is one selected from a group consisting of a piezoelectric element, a magnetostrictive actuator and a voice coil.
[claim4]
4. The 3-dimensional confocal microscopy apparatus according to any one of claims 1 to 3, wherein timing for driving the lens drive means for the pair of lenses for focal plane displacement and timing for acquiring an image by a fluorescent imaging camera are adjusted so as to be synchronous with each other.
[claim5]
5. The 3-dimensional confocal microscopy apparatus according to any one of claims 1 to 4, wherein an arithmetic operation using an arithmetic operation means for acquiring a 3-dimensional structural image of a specimen is performed by a Graphics Processing Unit (GPU).
[claim6]
6. The 3-dimensional confocal microscopy apparatus according to any one of claims 1 to 5, wherein the 3-dimensional confocal microscopy apparatus further comprises a beam splitter or a dichroic mirror which is arranged between the objective lens and the pair of lenses for focal plane displacement, and an imaging lens and a bright field imaging camera for imaging a 2-dimensional image of a specimen separated by the beam splitter or the dichroic mirror.
[claim7]
7. A focal plane scanning and aberration correction unit to be incorporated into a 3-dimensional confocal microscopy apparatus comprises
lens drive means (17) and
a pair of lenses (14, 15) for focal plane displacement, the pair of lenses for focal plane displacement being constituted of an intermediate imaging lens (14) which forms an intermediate image of a fluorescent confocal microscopy image of a specimen and an intermediate objective lens (15) arranged such that a focal point of the intermediate objective lens overlaps with a focal point of the intermediate imaging lens on the same optical axis, and at least one lens out of these lenses being movable in an optical axis direction by the lens drive means, and
the focal plane scanning and aberration correction unit further comprises a computer having an arithmetic operation means for acquiring a 3-dimensional structural image of the specimen by correcting aberration of the fluorescent confocal microscopy image caused by the movement of the lens of the pair of lenses for focal plane displacement in real time using the fluorescent confocal microscopy image acquired by the fluorescent light imaging camera and aberration correction data which is acquired in advance by performing provisional measurement of a sample specimen, and a means which displays a corrected image obtained by the arithmetic operation means,
wherein the 3-dimensional confocal microscopy apparatus comprises a light source for a laser (10); an objective lens (12) for focusing the laser supplied from the light source on a fluorescent specimen; and a fluorescent light imaging camera (16) for detecting a fluorescent confocal microscopy image based on a fluorescent light emitted from the fluorescent specimen, and wherein
the focal plane scanning and aberration correction unit is to be arranged between the objective lens and the fluorescent light imaging camera.
  • 出願人(英語)
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • IKUTA KOJI
  • IKEUCHI MASASHI
国際特許分類(IPC)
指定国 Contracting States: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
参考情報 (研究プロジェクト等) CREST Novel Measuring and Analytical Technology Contributions to the Elucidation and Application of Life Phenomena AREA
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