Light-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation - Archive ouverte HAL Access content directly
Journal Articles Optics Express Year : 2013

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

(1, 2) , (3, 4) , (3) , (5, 3) , (2, 6)
1
2
3
4
5
6

Abstract

In 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.
Fichier principal
Vignette du fichier
00994649.pdf (4.76 Mo) Télécharger le fichier
Origin : Publisher files allowed on an open archive

Dates and versions

hal-00994649 , version 1 (19-11-2015)

Identifiers

Cite

Florian Fahrbach, Vasily Gurchenkov, Kévin Alessandri, Pierre Nassoy, Alexander Rohrbach. Light-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation. Optics Express, 2013, 21 (11), pp.13824-13839. ⟨10.1364/OE.21.013824⟩. ⟨hal-00994649⟩
488 View
508 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More