Open AccessScanned light sheet microscopy with confocal slit detection
Author(s) -
Eugen Baumgart,
Ulrich Kubitscheck
Publication year - 2012
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.20.021805
Subject(s) - optical sectioning , light sheet fluorescence microscopy , optics , confocal , materials science , shutter , confocal microscopy , microscopy , rolling shutter , microscope , light scattering , optical microscope , scanning confocal electron microscopy , scattering , physics , scanning electron microscope
In light sheet fluorescence microscopy optical sectioning is achieved by illuminating the sample orthogonally to the detection pathway with a thin, focused sheet of light. However, light scattering within the sample often deteriorates the optical sectioning effect. Here, we demonstrate that contrast and degree of confocality can greatly be increased by combining scanned light sheet fluorescence excitation and confocal slit detection. A high frame rate was achieved by using the "rolling shutter" of a scientific CMOS camera as a slit detector. Synchronizing the "rolling shutter" with the scanned illumination beam results in confocal line detection. Acquiring image data with selective plane illumination minimizes photo-damage while simultaneously enhancing contrast, optical sectioning and signal-to-noise ratio. Thus the imaging principle presented here merges the benefits of scanned light sheet microscopy and line-scanning confocal imaging.