We have developed a light-sheet illumination microscope that can perform fast 3D imaging of transparent biological samples with inexpensive visible lasers and a single galvo mirror (GM). The light-sheet is created by raster scanning a Bessel beam with a GM, with this same GM also being used to rescan the fluorescence across a chip of a camera to construct an image in real time. A slit is used to reject out-of-focus fluorescence such that the image formed in real time has minimal contribution from the sidelobes of the Bessel beam. Compared with two-photon Bessel beam excitation or other confocal line-scanning approaches, our method is of lower cost, is simpler, and does not require calibration and synchronization of multiple GMs. We demonstrated the optical sectioning and out-of-focus background rejection capabilities of this microscope by imaging fluorescently labeled actin filaments in fixed 3T3 cells.
© 2014 Optical Society of America
Original Manuscript: March 13, 2014
Manuscript Accepted: May 9, 2014
Published: June 13, 2014
Vol. 9, Iss. 8 Virtual Journal for Biomedical Optics
P. Zhang, M. E. Phipps, P. M. Goodwin, and J. H. Werner, "Confocal line scanning of a Bessel beam for fast 3D Imaging," Opt. Lett. 39, 3682-3685 (2014)