WO2001061324A1 - Techniques de microscopie par fluorescence et dispositifs utilisant les diodes a emission de lumiere - Google Patents

Techniques de microscopie par fluorescence et dispositifs utilisant les diodes a emission de lumiere Download PDF

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Publication number
WO2001061324A1
WO2001061324A1 PCT/US2001/005107 US0105107W WO0161324A1 WO 2001061324 A1 WO2001061324 A1 WO 2001061324A1 US 0105107 W US0105107 W US 0105107W WO 0161324 A1 WO0161324 A1 WO 0161324A1
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WO
WIPO (PCT)
Prior art keywords
recited
microscope
sample
leds
radiation
Prior art date
Application number
PCT/US2001/005107
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English (en)
Inventor
Victor Barsky
Andrei Mirzabekov
Yuri Vengerov
Yegor Yergorov
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The University Of Chicago
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The University Of Chicago filed Critical The University Of Chicago
Priority to AU2001237056A priority Critical patent/AU2001237056A1/en
Publication of WO2001061324A1 publication Critical patent/WO2001061324A1/fr

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Classifications

    • 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
    • G02B21/008—Details of detection or image processing, including general computer control
    • 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
    • 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
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • 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
    • G01N2021/6417—Spectrofluorimetric devices
    • G01N2021/6419—Excitation at two or more wavelengths

Definitions

  • the present invention relates to a device and a method for illuminating samples in microscopy, and more particularly, the present invention relates to a device and a method for incorporating light emitting diodes in fluorescence microscopy procedures.
  • Fluorescence microscopy has many uses related to detection of target molecules.
  • Target molecules when illuminated with excitation light may either fluoresce spontaneously or they may fluoresce after a tag is attached thereto.
  • a fluorescence tag is attached onto a moiety which itself is complimentary to the topography or chemistry of the target molecule.
  • the fluorescent tag attached to the moiety indicates the existence of the duplex thus formed.
  • two light sources are used for fluorescent microscopy, namely lasers and high power lamps (such as tungsten-, mercury-, and xenon-lamps).
  • Lasers are employed mainly with scanning devices to provide a high power output in a narrow spectral region on a small surface. Lamps provide powerful illumination of large fields and also provide different wavelength capabilities.
  • Present microscope designs used in carrying out routine detection of fluorescent molecules incorporate epi-fluorescence microscopy. Epi-fluorescence microscopy most frequently employs high power tungsten or arc lamps in combination with light filters to excite the fluorescence in the sample.
  • Epi- is a reference to the so-called epi-illumination scheme wherein the same objective lens is used for both illuminating the sample and collecting the fluorescence.
  • These systems require the use of high-powered mercury or xenon lamps to ensure that after narrow-band filtering (monochro-matization), there is still enough light to provide a measurable signal at the detector.
  • these sources are inefficient in that they produce a large amount of heat and light in excess of that required to illuminate the sample.
  • the large power and size requirements of these lamps makes systems incorporating them bulky. So, portability is a problem. Costs for these systems range from $12,000 to $100,000.
  • Lasers have been utilized in fluorescence microscopy processes. Elaborate confocal microscopy methods (i.e., confocal laser scanners) and devices exist (e.g. U.S. Patent Nos. 5,631 ,734 and 5,578,832) whereby sharply focused laser beams scan a sample point by point. Lasers emit light in a narrow spectral region, and as such, obviate the need for polarization filters.
  • fluorochromes which excite at different wave lengths, different lasers must be employed, leading to higher costs. Also, lasers do not provide enough power to illuminate the whole microscope field.
  • confocal laser systems rely on complex, and therefore expensive, electronics (e.g., photomultiplier detectors and digitizing devices) to reconstruct the image as a whole. Indeed, confocal laser scanning systems cost from $500,000 to $1 million. Unlike epi-fluorescence microscopes, confocal configurations do not record the fluorescing image in parallel (i.e., all points at once).
  • An exemplary state of the art confocal configuration is depicted in FIG. 1 as numeral 10. Briefly, a stationary laser light source 12 supplies a light beam 14 for illuminating an objective 16. The beam is required to traverse a complex mirror- and piezo quartz-configuration 20 before arriving at the objective.
  • the fluorescence energy must traverse through the dichroic mirror 20 to reach a detector 22 such as a photo multiplier tube.
  • a detector 22 such as a photo multiplier tube.
  • a confocal pinhole 24 is juxtaposed intermediate to the objective and the detector.
  • a signal processing unit 26 delivers the fluorescence signal to a computer 28 for processing. This configuration requires that the sample be able to move with respect to the laser (or, conversely, that the laser be able to move with respect to the sample).
  • Still another object of the present invention is to provide a low-cost alternative to typical fluorescence microscopy configurations.
  • a feature of the device is the illumination of an entire sample and at an angle, or by some other means, which precludes exposure of the objective to incident or reflected illuminating radiation.
  • An advantage of the device is that exposure of the detector to reflected light is eliminated without the need for dichroic mirrors, confocal pinholes, or other means to shield the detector.
  • Yet another object of the present invention is to provide an efficient, compact, and portable fluorescence microscopy method.
  • a feature of the invention is illuminating the sample with modular, low-energy (and therefore low-heat) LEDs that can be battery powered.
  • Another feature of the invention is that the LED provides a swath of light that can be positioned to illuminate the entire sample at once.
  • An advantage of the method is that the simultaneous illumination of the entire sample facilitates parallel analysis of the sample located in different regions of the field of view (FOV).
  • Another advantage is the ability to substitute, or use simultaneously, different LEDs so as to either serially- or simultaneously-illuminate the same sample with different radiation.
  • the invention provides for a device for fluorescence microscopy comprising a means for illuminating a sample undergoing microscopic examination by juxtaposing a appropriately filtered light from one or more light emitting diode to the sample so as to irradiate the sample, a means for filtering and detecting radiation fluorescing from the sample and visually monitoring or permanently imaging the radiation from the sample, and a means for preventing said detecting means from detecting non-fluorescing radiation from the sample or fluorescent radiation that does not originate from the sample.
  • the invention also provides for a method for illuminating a microscopic construct, the method comprising providing an illumination detector capable of simultaneously monitoring all points of the construct; and simultaneously directing illuminating radiation to all regions of the construct and in a manner that prevents exposure of the detector to the illuminating radiation.
  • FIG. 1 is a schematic depiction of a typical state-of-the-art confocal fluorescence microscope configuration
  • FIG. 2 is a schematic depiction of an exemplary fluorescence microscope to provide dark-field epi-illumination using LEDs in combination with a ring mirror, in accordance with features of the present invention
  • FIG. 3 is a schematic depiction of another fluorescence microscope but without a ring mirror, in accordance with features of the present invention
  • FIG. 4a is a schematic depiction of a means for providing sample illumination using a fiber optic cable, in accordance with features of the present invention
  • FIG. 4b is a schematic depiction of a means for providing uniform sample illumination using fiber optic cables, in accordance with features of the present invention
  • FIG. 8 is a cross sectional view of an LED embedded into an objective, in accordance with features of the present invention.
  • FIG. 9 is a schematic view of a sample illumination configuration utilizing two parabolic mirrors, in accordance with features of the present invention.
  • the LEDs may be located inside the microscope, between the objective lens and the image detector, one possible configuration being a ring of LEDs coaxial with the microscope, the LEDs themselves being surrounded by an annular mirror that focuses the LED light onto the sample. Or the LED's light may be transmitted to the sample by means of optical fibers terminating in the region between the objective lens and the detector.
  • the LEDs small size, high power output, low heat output, high quantum yield, and their interchangeability allows one to design devices that cannot be built with other light sources.
  • the use of LEDs results in fluorescence microscopes of smaller dimensions, simpler construction, better signal to noise ratio and higher sample illumination, 3mW/mm as compared to 1mW/mm 2 with other light sources.
  • the sheath instead of a frusto-conical configuration, the sheath defines a parabolic section. As with the frusto-conical section described supra, the parabolic section defines inwardly facing surfaces converging toward the sample.
  • the sheath is configured to direct light to the sample so as to prevent any light reflecting off of the sample from being read by the detector 68.
  • the reflective surface of the sheath is configured so that light incident upon the reflective surface is directed at an angle toward the sample so as to prevent reflection of un-utilized excitation light into the field of view of the detector. Thus, only light emanating from the sample as a result of fluorescence of the sample is detected.
  • Utilizing the sheath also eliminates the state-of-the art requirement of illuminating the sample through an objective 63. As such, no optics fluorescence occurs and therefore a further decrease in background radiation is realized. Increased sensitivity and contrast results from this invented arrangement. This embodiment has become practical now that high output LEDs have become available.
  • the fluorescence emanating from the illuminated sample 64 is observed via an eye piece 68 or recorded via a detector, such as photographic film, CCD camera, etc..
  • the light beam 56 impinges on the sample 64 for a time sufficient to induce the excitation of fluorochromes located in the sample
  • one or a plurality of LEDs 52 are optionally situated inside the sheath, as depicted in FIG. 3.
  • the reflective surface is configured to direct light to the sample 64 at an angle ⁇ so as to prevent any of the directed light, or any reflecting portion of the directed light to be read by a fluorescence-recording or -detecting means 68.
  • an impinging light beam or reflected light beam which does not travel in parallel with the focal path of the microscope indicates that the LED and reflective surfaces are suitably oriented, relative to each other.
  • a typical configuration therefore is side illumination of the sample, i.e, where an angularly and medially-directed illumination light is applied to the sample from a point lateral from the longitudinal axis of the microscope.
  • Such a configuration obviates the need for dichroic mirrors, confocal pinholes or other filtering devices.
  • the light radiation from the LED may first be directed to an optofiber system comprising an axial optical fiber 70 as depicted in FIG. 4a or a plurality of optical fibers 71 as depicted in FIG. 4b.
  • Suitable optical fibers are available from a myriad of commercial suppliers, including Optical Cable Corporation of Roanoke, Virginia, USA, and Litkarino, of Moscow, Russia.
  • FIG. 5 depicts the utilization of LEDs in a dark-field configuration.
  • this configuration provides impinging radiation 57 at a non-perpendicular angle to the sample.
  • the sample is illuminated in a dark field wherein the transmitted light beams impinge upon the sample at a point lateral from the midline 65 of the field of view. That the impinging radiation contacts the sample 64 at an angle not perpendicular to the plane 72 upon which the sample rests is achieved via a cardioid, parabolic, or dark-centered condenser 73 juxtaposed intermediate the LED 52 and the sample 64.
  • the LED and the condenser are located distal from the detector relative to the sample 64 and below the plane 72 of the sample.
  • the arrangement depicted in FIG. 5 is particularly suited for viewing transparent samples mounted on transparent supports.
  • This arrangement has the advantage that the exciting radiation does not impinge on the objective 63 and thus does not contribute to the background. Best results are obtained when the excitation beam is formed with the help of a parabolic condenser positioned between the LED 52 and the sample 64.
  • the inventors have utilized the invented fluorescence microscopy configuration to illuminate tagged moieties residing in nearly microscopic acrylamide substrates (0.1 x 0.1 x 0.02 mm). In such scenarios, wherein as little as 5 x 10 5 molecules of Texas red may be detected with average excitation intensities of between 0.5 and 10 mW/mm 2 .
  • a myriad of types of light emitting diodes lasers are utilized in the invented device.
  • a suitable laser must emit a radiation suitable to induce fluorescence from typical chromophores used in tagging processes.
  • Typical chromophores include, but are not limited to, Texas Red, Cy dyes, Naphthofluorescein, Fluorescein, BODIPY dyes and others.
  • Table 1 shows the various types of fluorochromes that can be utilized with various LEDs currently available.
  • FIG. 6 A perspective view of the LED-illuminated objective, is depicted in FIG. 6. Also depicted in FIG. 6 is a power-source 75 removably attached to a region of the objective's housing 76. The power source is provided for energizing the LEDs. For the sake of convenience, the LEDs typically are removably, and circumferentially arranged along the periphery of the objective's housing 76.
  • the LEDs can be permanently affixed to the objective housing with adhesive, mechanical fasteners, or integrally molded to the material forming said housing.
  • This scheme allows design and use of specialized objectives which can be incorporated into an ordinary microscope and transforming it into a fluorescence microscope.
  • the fluorescent lighting may be used instead of or in conjunction with ordinary microscope lights.
  • the intensity of illumination inside the field of view of the 20 x 0.5 objective was approximately 3 mW/sq. mm. This intensity is used to excite FITC, which is an activated derivative of fluorescein.
  • FITC an activated derivative of fluorescein.
  • exposure time for tubercle bacilli immersed with fluorescein, and washed was approximately 2-3 seconds. This exposure time is comparable with those provided by commercially-available fluoresc- ence microscopes.
  • FIG. 9 depicts an illumination configuration utilizing two parabolic mirrors.
  • the sample, 64 is directly illuminated by a light source 52.
  • the light source directs energizing radiation away from the detector 68. This radiation from the light source first passes through a lens 54.
  • the first parabolic mirror 80 is concentrically arranged about the longitudinal axis ⁇ of the microscope with the surface of the mirror deviating medially, or inwardly toward the axis.
  • the parabolic configuration of the first reflective surface depends downwardly toward a plane 86 supporting the sample.
  • the first and second reflective surfaces are optimized in albedo and configuration to maximize capture of fluorescing radiation emanating from the sample.
  • An advantage of the dual reflectance configuration of FIG. 9 is the complete isolation of the detector 68 from non-fluorescing radiation emanating from the light source 52.
  • a backstop 88 of the light source, a backstop 90 of the first reflective surface 80 and a backstop 92 of the second reflective surface 82 are configured to be opaque to the energizing radiation utilized in the device depicted in FIG. 9. This scheme is made possible by the present availability of small-size LEDs together with large size mirror objectives.
  • the light source 52 need not be incorporated into the dual mirror configuration.
  • one or a plurality of optical fibers 70 can illuminate the sample 64 by directing light to the sample from an LED 52 or plurality of LEDs not embodied by the device.
  • significant increase in the illumination intensity may be achieved by having the fluorescence radiation be brought to the sample by optical cables and by placing the end of the cables near the sample.

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  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Optics & Photonics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
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  • Engineering & Computer Science (AREA)
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  • General Engineering & Computer Science (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

La présente invention concerne un dispositif permettant d'illuminer un échantillon (64) soumis à un examen microscopique, ce dispositif comprenant un détecteur (68) de rayonnement fluorescent (68) et une diode (52) émettrice de lumière juxtaposée à cet échantillon de façon à irradier cet échantillon et empêcher l'organe de détection de détecter un rayonnement non fluorescent émanant de cet échantillon.
PCT/US2001/005107 2000-02-17 2001-02-16 Techniques de microscopie par fluorescence et dispositifs utilisant les diodes a emission de lumiere WO2001061324A1 (fr)

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Application Number Priority Date Filing Date Title
AU2001237056A AU2001237056A1 (en) 2000-02-17 2001-02-16 Fluorescence microscopy methods and devices using light emission diodes

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Application Number Priority Date Filing Date Title
RU2000104231/28A RU2182328C2 (ru) 2000-02-17 2000-02-17 Флуоресцентный микроскоп
RU2000104231 2000-02-17

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Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003073079A2 (fr) * 2002-02-22 2003-09-04 Xenogen Corporation Ensemble d'eclairage fluorescent pour appareil d'imagerie
WO2004088387A1 (fr) * 2003-04-02 2004-10-14 Fraen Corporation S.R.L. Ensemble d’eclairage pour appareil d’analyse par luminescence, notamment un microscope a fluorescence, et appareil d’analyse par luminescence equipe d’un tel ensemble d’eclairage
DE10314125A1 (de) * 2003-03-28 2004-10-21 Carl Zeiss Jena Gmbh Anordnung zur Beleuchtung von Objekten mit Licht unterschiedlicher Wellenlänge
US6922246B2 (en) 2002-02-22 2005-07-26 Xenogen Corporation Bottom fluorescence illumination assembly for an imaging apparatus
NL1027627C2 (nl) * 2004-11-30 2006-05-31 Ccm Beheer Bv Verlichtingssysteem.
WO2006077304A1 (fr) * 2005-01-21 2006-07-27 Cypher Science Appareil de detection portable permettant de detecter sur le terrain des elements marques par fluorescence
WO2007011818A1 (fr) * 2005-07-15 2007-01-25 Auburn University Dispositif d'eclairage pour microscope et adaptateur d'eclairage a fond noir et a fond clair
US7383078B2 (en) 2000-02-25 2008-06-03 Xenogen Corporation Imaging apparatus with heating element
WO2008095880A1 (fr) * 2007-02-09 2008-08-14 Leica Microsystems (Schweiz) Ag Dispositif d'éclairage pour un microscope
EP1963827A1 (fr) * 2005-10-26 2008-09-03 Kabushiki Kaisha Toshiba Appareil d'analyse
FR2913499A1 (fr) * 2007-03-09 2008-09-12 Genewave Soc Par Actions Simpl Dispositif de lecture de fluorescence.
US7474398B2 (en) 2002-02-22 2009-01-06 Xenogen Corporation Illumination system for an imaging apparatus with low profile output device
US7474399B2 (en) 2002-02-22 2009-01-06 Xenogen Corporation Dual illumination system for an imaging apparatus and method
WO2011098811A1 (fr) * 2010-02-12 2011-08-18 University Of Northumbria At Newcastle Appareil d'émission et de réception de rayonnement
EP1549988B2 (fr) † 2002-10-08 2011-10-05 Karl Kaps GmbH & Co KG Dispositif d'eclairage pour un instrument d'agrandissement optique et instrument d'agrandissement optique
GB2483963A (en) * 2010-08-27 2012-03-28 Univ Leland Stanford Junior Epifluorescence microscope with low power input
US8901516B2 (en) 2010-09-01 2014-12-02 Spectral Instruments Imaging, LLC Excitation light source assembly
EP2896957A1 (fr) * 2014-01-21 2015-07-22 Molecular Devices, LLC Système de détection à base de filtre et à base de monochromateur
US9347894B2 (en) 2010-09-01 2016-05-24 Spectral Instruments Imaging, LLC Methods and systems for producing visible light and x-ray image data
EP2246691B1 (fr) * 2009-04-29 2016-07-13 Commissariat à l'Énergie Atomique et aux Énergies Alternatives Procédé et appareil de comptage des Thrombocytes
WO2019178090A1 (fr) * 2018-03-12 2019-09-19 The University Of North Carolina At Chapel Hill Microscopie à disque optique pour microscopes à fluorescence
US11099370B2 (en) 2016-09-09 2021-08-24 The University Of North Carolina At Chapel Hill Tilted illumination systems for fluorescence microscopes
US11262570B2 (en) 2018-03-12 2022-03-01 The University Of North Carolina At Chapel Hill Mirror image microscopy for increased collection

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1782432A1 (fr) * 2004-05-31 2007-05-09 Givargizov, Mikhail Evgenievich Structure de pointe pour dispositifs de balayage, procede permettant de produire cette structure et dispositifs comprenant cette structure
RU2348695C2 (ru) 2006-05-23 2009-03-10 Закрытое акционерное общество "Молекулярно-медицинские технологии" Дифференцирующий и специфический олигонуклеотиды для идентификации последовательностей днк инфекционных агентов в биологических материалах, способ видовой идентификации инфекционных агентов, биочип и набор для осуществления этого способа
WO2009002225A2 (fr) * 2007-06-25 2008-12-31 Closed Company 'molecular-Medicine Technologies' Dispositif multifonctions de diagnostic et procédé de test pour objets biologiques
EP2533033A4 (fr) * 2010-02-03 2013-07-10 Uchrezhdenie Rossiiskoi Akademii Nauk Inst Molekulyarnoi Biolog Im V A Engelgardta Ran Imb Ran Dispositif pour l'analyse de micropuces biologiques luminescentes

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3705755A (en) * 1970-08-24 1972-12-12 Stephen Charles Baer Microscopy apparatus
US5646411A (en) * 1996-02-01 1997-07-08 Molecular Dynamics, Inc. Fluorescence imaging system compatible with macro and micro scanning objectives
US6008892A (en) * 1997-05-23 1999-12-28 Molecular Dynamics, Inc. Optical substrate for enhanced detectability of fluorescence
US6154282A (en) * 1998-10-26 2000-11-28 Cytotelesis Inc. Semiconductor based excitation illuminator for fluorescence and phosphorescence microscopy

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3705755A (en) * 1970-08-24 1972-12-12 Stephen Charles Baer Microscopy apparatus
US5646411A (en) * 1996-02-01 1997-07-08 Molecular Dynamics, Inc. Fluorescence imaging system compatible with macro and micro scanning objectives
US6008892A (en) * 1997-05-23 1999-12-28 Molecular Dynamics, Inc. Optical substrate for enhanced detectability of fluorescence
US6154282A (en) * 1998-10-26 2000-11-28 Cytotelesis Inc. Semiconductor based excitation illuminator for fluorescence and phosphorescence microscopy

Cited By (48)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7383078B2 (en) 2000-02-25 2008-06-03 Xenogen Corporation Imaging apparatus with heating element
US7949383B2 (en) 2000-02-25 2011-05-24 Xenogen Corporation Imaging apparatus with selectable moveable stage
WO2003073079A2 (fr) * 2002-02-22 2003-09-04 Xenogen Corporation Ensemble d'eclairage fluorescent pour appareil d'imagerie
WO2003073079A3 (fr) * 2002-02-22 2004-01-22 Xenogen Corp Ensemble d'eclairage fluorescent pour appareil d'imagerie
US7474398B2 (en) 2002-02-22 2009-01-06 Xenogen Corporation Illumination system for an imaging apparatus with low profile output device
US7474399B2 (en) 2002-02-22 2009-01-06 Xenogen Corporation Dual illumination system for an imaging apparatus and method
US7466418B2 (en) 2002-02-22 2008-12-16 Xenogen Corporation Fluorescence illumination assembly for an imaging apparatus and method
US6894289B2 (en) 2002-02-22 2005-05-17 Xenogen Corporation Fluorescence illumination assembly for an imaging apparatus
US6922246B2 (en) 2002-02-22 2005-07-26 Xenogen Corporation Bottom fluorescence illumination assembly for an imaging apparatus
US7177024B2 (en) 2002-02-22 2007-02-13 Xenogen Corporation Bottom fluorescence illumination assembly for an imaging apparatus
EP1549988B2 (fr) † 2002-10-08 2011-10-05 Karl Kaps GmbH & Co KG Dispositif d'eclairage pour un instrument d'agrandissement optique et instrument d'agrandissement optique
DE10314125B4 (de) * 2003-03-28 2005-02-24 Carl Zeiss Jena Gmbh Anordnung zur Beleuchtung von Objekten mit Licht unterschiedlicher Wellenlänge
DE10314125A1 (de) * 2003-03-28 2004-10-21 Carl Zeiss Jena Gmbh Anordnung zur Beleuchtung von Objekten mit Licht unterschiedlicher Wellenlänge
US7416313B2 (en) 2003-03-28 2008-08-26 Carl Zeiss Microimaging Gmbh Assembly for illuminating objects with light of different wavelengths
WO2004088387A1 (fr) * 2003-04-02 2004-10-14 Fraen Corporation S.R.L. Ensemble d’eclairage pour appareil d’analyse par luminescence, notamment un microscope a fluorescence, et appareil d’analyse par luminescence equipe d’un tel ensemble d’eclairage
WO2006059900A1 (fr) * 2004-11-30 2006-06-08 C.C.M. Beheer B.V. Système d’éclairage
NL1027627C2 (nl) * 2004-11-30 2006-05-31 Ccm Beheer Bv Verlichtingssysteem.
US7795598B2 (en) 2005-01-21 2010-09-14 Cypher Science Portable detection device for detecting on the ground elements marked by fluorescence
WO2006077304A1 (fr) * 2005-01-21 2006-07-27 Cypher Science Appareil de detection portable permettant de detecter sur le terrain des elements marques par fluorescence
FR2881225A1 (fr) * 2005-01-21 2006-07-28 Cypher Science Sarl Appareil de detection portable permettant de detecter sur le terrain des elements marques par fluorescence
WO2007011818A1 (fr) * 2005-07-15 2007-01-25 Auburn University Dispositif d'eclairage pour microscope et adaptateur d'eclairage a fond noir et a fond clair
US7542203B2 (en) 2005-07-15 2009-06-02 Auburn University Microscope illumination device and adapter
EP1963827A4 (fr) * 2005-10-26 2013-01-23 Toshiba Kk Appareil d'analyse
EP1963827A1 (fr) * 2005-10-26 2008-09-03 Kabushiki Kaisha Toshiba Appareil d'analyse
WO2008095880A1 (fr) * 2007-02-09 2008-08-14 Leica Microsystems (Schweiz) Ag Dispositif d'éclairage pour un microscope
US8331020B2 (en) 2007-02-09 2012-12-11 Leica Instruments (Singapore) Pte. Ltd. Illumination device for a microscope
WO2008132325A3 (fr) * 2007-03-09 2009-01-08 Genewave Dispositif de lecture de fluorescence
WO2008132325A2 (fr) * 2007-03-09 2008-11-06 Genewave Dispositif de lecture de fluorescence
FR2913499A1 (fr) * 2007-03-09 2008-09-12 Genewave Soc Par Actions Simpl Dispositif de lecture de fluorescence.
EP2246691B1 (fr) * 2009-04-29 2016-07-13 Commissariat à l'Énergie Atomique et aux Énergies Alternatives Procédé et appareil de comptage des Thrombocytes
WO2011098811A1 (fr) * 2010-02-12 2011-08-18 University Of Northumbria At Newcastle Appareil d'émission et de réception de rayonnement
EP2359744A1 (fr) * 2010-02-12 2011-08-24 University of Northumbria at Newcastle Appareil d'émission et de réception de radiation
US9498135B2 (en) 2010-08-27 2016-11-22 The Board Of Trustees Of The Leland Stanford Junior University Microscopy imaging device with advanced imaging properties
GB2483963A (en) * 2010-08-27 2012-03-28 Univ Leland Stanford Junior Epifluorescence microscope with low power input
US10813552B2 (en) 2010-08-27 2020-10-27 The Board Of Trustees Of The Leland Stanford Junior University Microscopy imaging device with advanced imaging properties
US9195043B2 (en) 2010-08-27 2015-11-24 The Board Of Trustees Of The Leland Stanford Junior University Microscopy imaging device with advanced imaging properties
GB2483963B (en) * 2010-08-27 2015-02-04 Univ Leland Stanford Junior Microscopy imaging device with advanced imaging properties
US9629554B2 (en) 2010-08-27 2017-04-25 The Board Of Trustees Of The Leland Stanford Junior University Microscopy imaging device with advanced imaging properties
US9474448B2 (en) 2010-08-27 2016-10-25 The Board Of Trustees Of The Leland Stanford Junior University Microscopy imaging device with advanced imaging properties
US9347894B2 (en) 2010-09-01 2016-05-24 Spectral Instruments Imaging, LLC Methods and systems for producing visible light and x-ray image data
US8901516B2 (en) 2010-09-01 2014-12-02 Spectral Instruments Imaging, LLC Excitation light source assembly
EP3293510A1 (fr) * 2014-01-21 2018-03-14 Molecular Devices, LLC Système de microscopie fluorescente
EP2896957A1 (fr) * 2014-01-21 2015-07-22 Molecular Devices, LLC Système de détection à base de filtre et à base de monochromateur
US11099370B2 (en) 2016-09-09 2021-08-24 The University Of North Carolina At Chapel Hill Tilted illumination systems for fluorescence microscopes
WO2019178090A1 (fr) * 2018-03-12 2019-09-19 The University Of North Carolina At Chapel Hill Microscopie à disque optique pour microscopes à fluorescence
CN112005100A (zh) * 2018-03-12 2020-11-27 查珀尔希尔北卡罗来纳大学 用于荧光显微镜的光盘显微术
US11262570B2 (en) 2018-03-12 2022-03-01 The University Of North Carolina At Chapel Hill Mirror image microscopy for increased collection
US11314074B2 (en) 2018-03-12 2022-04-26 The University Of North Carolina At Chapel Hill Light disc microscopy for fluorescence microscopes

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