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1. WO2011059826 - MULTIPLE-PHOTON EXCITATION LIGHT SHEET ILLUMINATION MICROSCOPE

Publication Number WO/2011/059826
Publication Date 19.05.2011
International Application No. PCT/US2010/054760
International Filing Date 29.10.2010
IPC
G02B 21/06 2006.1
GPHYSICS
02OPTICS
BOPTICAL ELEMENTS, SYSTEMS, OR APPARATUS
21Microscopes
06Means for illuminating specimen
G02B 21/00 2006.1
GPHYSICS
02OPTICS
BOPTICAL ELEMENTS, SYSTEMS, OR APPARATUS
21Microscopes
G01N 21/64 2006.1
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
CPC
G01N 2021/655
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
65Raman scattering
653Coherent methods [CARS]
655Stimulated Raman
G01N 21/6408
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6408with measurement of decay time, time resolved fluorescence
G01N 21/6428
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
G01N 21/6458
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
645Specially adapted constructive features of fluorimeters
6456Spatial resolved fluorescence measurements; Imaging
6458Fluorescence microscopy
G01N 21/6486
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6486Measuring fluorescence of biological material, e.g. DNA, RNA, cells
G02B 21/002
GPHYSICS
02OPTICS
BOPTICAL ELEMENTS, SYSTEMS, OR APPARATUS
21Microscopes
0004specially adapted for specific applications
002Scanning microscopes
Applicants
  • CALIFORNIA INSTITUTE OF TECHNOLOGY [US]/[US] (AllExceptUS)
  • TRUONG, Thai, V. [US]/[US] (UsOnly)
  • CHOI, John, M. [US]/[US] (UsOnly)
  • FRASER, Scott, E. [US]/[US] (UsOnly)
  • SUPATTO, Willy [FR]/[FR] (UsOnly)
  • KOOS, David, S. [US]/[US] (UsOnly)
Inventors
  • TRUONG, Thai, V.
  • CHOI, John, M.
  • FRASER, Scott, E.
  • SUPATTO, Willy
  • KOOS, David, S.
Agents
  • PECK, John, W.
Priority Data
61/256,00529.10.2009US
61/256,01029.10.2009US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) MULTIPLE-PHOTON EXCITATION LIGHT SHEET ILLUMINATION MICROSCOPE
(FR) MICROSCOPE À FEUILLE DE LUMIÈRE PAR ILLUMINATION ET EXCITATION MULTIPHOTON
Abstract
(EN)
An apparatus for and method of performing multi-photon light sheet microscopy (MP-LISH), combining multi-photon excited fluorescence with the orthogonal illumination of light sheet microscopy are provided. With live imaging of whole Drosophila and zebrafish embryos, the high performance of MP-LISH compared to current state-of-the-art imaging techniques in maintaining good signal and high spatial resolution deep inside biological tissues (two times deeper than one-photon light sheet microscopy), in acquisition speed (more than one order of magnitude faster than conventional two-photon laser scanning microscopy), and in low phototoxicity are demonstrated. The inherent multi-modality of this new imaging technique is also demonstrated second harmonic generation light sheet microscopy to detect collagen in mouse tail tissue. Together, these properties create the potential for a wide range of applications for MP-LISH in 4D imaging of live biological systems.
(FR)
L'invention concerne un appareil et un procédé de réalisation de la microscopie à feuille de lumière multiphotonique, ce procédé combinant la fluorescence par excitation multiphoton avec l'illumination orthogonale de la microscopie à feuille de lumière. Par l'imagerie d'embryons complets vivants de Drosophiles et de poissons zèbre, il a été démontré la haute performance du microscope à feuille de lumière multiphoton en comparaison avec des techniques d'imagerie courantes de pointe, pour ce qui est du maintien d'un bon signal et d'une haute résolution spatiale en profondeur à l'intérieur des tissus biologiques (deux fois plus profond que la microscopie à feuille de lumière monophotonique), de la vitesse d'acquisition d'image (d'un ordre de grandeur plus rapide que la microscopie à balayage laser à deux photons conventionnelle) et d'une faible toxicité. La multimodalité inhérente de cette nouvelle technique d'imagerie a également été démontrée par la microscopie à feuille de lumière par génération de seconde harmonique afin de rechercher le collagène dans le tissu d'une queue de souris. Toutes ces propriétés réunies créent le potentiel d'une large plage d'applications pour la microscopie à feuille de lumière multiphotonique dans l'imagerie 4D de systèmes biologiques vivants.
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