Simple imaging protocol for autofluorescence elimination and optical sectioning in fluorescence endomicroscopy

Our Research / Publications

janelia7_blocks-janelia7_biblio_header | block

Optica. 2019 Jul;6:972. doi: 10.1364/optica.6.000972

Simple imaging protocol for autofluorescence elimination and optical sectioning in fluorescence endomicroscopy Tebo Lab

Zhang R, Chouket R, Tebo AG, Plamont M, Kelemen Z, Gissot L, Faure J, Gautier A, Croquette V, Jullien L, Le Saux TL

Note: Research in this publication was not performed at Janelia.

janelia7_blocks-janelia7_biblio_abstract | block

Abstract

Fiber-optic epifluorescence imaging with one-photon excitation benefits from its ease of use, cheap light sources, and full-frame acquisition, which enables it for favorable temporal resolution of image acquisition. However, it suffers from a lack of robustness against autofluorescence and light scattering. Moreover, it cannot easily eliminate the out-of-focus background, which generally results in low-contrast images. In order to overcome these limitations, we have implemented fast out-of-phase imaging after optical modulation (Speed OPIOM) for dynamic contrast in fluorescence endomicroscopy. Using a simple and cheap optical-fiber bundle-based endomicroscope integrating modulatable light sources, we first showed that Speed OPIOM provides intrinsic optical sectioning, which restricts the observation of fluorescent labels at targeted positions within a sample. We also demonstrated that this imaging protocol efficiently eliminates the interference of autofluorescence arising from both the fiber bundle and the specimen in several biological samples. Finally, we could perform multiplexed observations of two spectrally similar fluorophores differing by their photoswitching dynamics. Such attractive features of Speed OPIOM in fluorescence endomicroscopy should find applications in bioprocessing, clinical diagnostics, plant observation, and surface imaging.

node:body | entity_field

Main Menu (Mobile)- Block

Main Menu - Block

Simple imaging protocol for autofluorescence elimination and optical sectioning in fluorescence endomicroscopy

Note: Research in this publication was not performed at Janelia.

Abstract

Janelia Authors