title: Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.
abstract: The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume.
authors.affiliation: Department of Electrical and Computer Engineering, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
authors.forename: Qian
authors.initials: Q
authors.lastname: Ma
authors.affiliation: Department of Electrical and Computer Engineering, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
authors.forename: Bahar
authors.initials: B
authors.lastname: Khademhosseinieh
authors.affiliation: Department of Physics, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Huang
authors.affiliation: Department of Electrical and Computer Engineering, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
authors.forename: Haoliang
authors.initials: H
authors.lastname: Qian
authors.affiliation: Division of Biological Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
authors.forename: Malina A
authors.initials: MA
authors.lastname: Bakowski
authors.affiliation: Division of Biological Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
authors.forename: Emily R
authors.initials: ER
authors.lastname: Troemel
authors.affiliation: Department of Electrical and Computer Engineering, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
authors.forename: Zhaowei
authors.initials: Z
authors.lastname: Liu