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Light-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation

hal.structure.identifierMax Planck Institute of Molecular Cell Biology and Genetics [MPI-CBG]
hal.structure.identifierLaboratory for Bio- and Nano-Photonics, University of Freiburg
dc.contributor.authorFAHRBACH, Florian
hal.structure.identifierPhysico-Chimie-Curie [PCC]
hal.structure.identifierCompartimentation et dynamique cellulaires [CDC]
dc.contributor.authorGURCHENKOV, Vasily
hal.structure.identifierPhysico-Chimie-Curie [PCC]
dc.contributor.authorALESSANDRI, Kévin
hal.structure.identifierlp2n-04,lp2n-16
hal.structure.identifierPhysico-Chimie-Curie [PCC]
dc.contributor.authorNASSOY, Pierre
hal.structure.identifierLaboratory for Bio- and Nano-Photonics, University of Freiburg
hal.structure.identifierCentre for Biological Signalling Studies (bioss)
dc.contributor.authorROHRBACH, Alexander
dc.date.accessioned2023-05-12T10:58:46Z
dc.date.available2023-05-12T10:58:46Z
dc.date.issued2013-06-03
dc.identifier.issn1094-4087
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/181983
dc.description.abstractEnIn this study we show that it is possible to successfully combine the benefits of light-sheet microscopy, self-reconstructing Bessel beams and two-photon fluorescence excitation to improve imaging in large, scattering media such as cancer cell clusters. We achieved a nearly two-fold increase in axial image resolution and 5-10 fold increase in contrast relative to linear excitation with Bessel beams. The light-sheet penetration depth could be increased by a factor of 3-5 relative to linear excitation with Gaussian beams. These findings arise from both experiments and computer simulations. In addition, we provide a theoretical description of how these results are composed. We investigated the change of image quality along the propagation direction of the illumination beams both for clusters of spheres and tumor multicellular spheroids. The results reveal that light-sheets generated by pulsed near-infrared Bessel beams and two photon excitation provide the highest image resolution, contrast and light-sheet penetration at a minimum amount of artifacts.
dc.language.isoen
dc.publisherOptical Society of America - OSA Publishing
dc.subject.enDEEP
dc.subject.enFIELD
dc.subject.enSELECTIVE PLANE ILLUMINATION
dc.subject.enPROPAGATION
dc.subject.enDEPTH
dc.title.enLight-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation
dc.typeArticle de revue
dc.identifier.doi10.1364/OE.21.013824
dc.subject.halPhysique [physics]/Physique [physics]/Biophysique [physics.bio-ph]
dc.subject.halPhysique [physics]/Physique [physics]/Optique [physics.optics]
bordeaux.journalOptics Express
bordeaux.page13824-13839
bordeaux.volume21
bordeaux.hal.laboratoriesLaboratoire Photonique, Numérique et Nanosciences (LP2N) - UMR 5298*
bordeaux.issue11
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionCNRS
bordeaux.peerReviewedoui
hal.identifierhal-00994649
hal.version1
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-00994649v1
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Optics%20Express&rft.date=2013-06-03&rft.volume=21&rft.issue=11&rft.spage=13824-13839&rft.epage=13824-13839&rft.eissn=1094-4087&rft.issn=1094-4087&rft.au=FAHRBACH,%20Florian&GURCHENKOV,%20Vasily&ALESSANDRI,%20K%C3%A9vin&NASSOY,%20Pierre&ROHRBACH,%20Alexander&rft.genre=article


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