Article abstract
Nature Photonics 2, 190 - 195 (2008)
Published online: 17 February 2008 | doi:10.1038/nphoton.2007.300
Subject Categories: Imaging and sensing | Biophotonics
Non-scanning motionless fluorescence three-dimensional holographic microscopy
Joseph Rosen1,2,3 & Gary Brooker1,2,3
Abstract
Holography is an attractive imaging technique as it offers the ability to view a complete three-dimensional volume from one image. However, holography is not widely applied to the field of three-dimensional fluorescence microscopic imaging, because fluorescence is incoherent and creating holograms requires a coherent interferometer system. Although scanning one beam of an interferometer pattern across the rear aperture of an objective to excite fluorescence in a specimen overcomes the coherence limitation, the mechanical scanning is complicated, which makes the image capturing slow, and the process is limited to low-numerical-aperture objectives. Here we present the first demonstration of a motionless microscopy system (FINCHSCOPE) based on Fresnel incoherent correlation holography, and its use in recording high-resolution three-dimensional fluorescent images of biological specimens. By using high-numerical-aperture objectives, a spatial light modulator, a CCD camera and some simple filters, FINCHSCOPE enables the acquisition of three-dimensional microscopic images without the need for scanning.
- Johns Hopkins University Microscopy Center, Montgomery County Campus 9605 Medical Center Drive Suite 240, Rockville, Maryland 20850, USA
- Johns Hopkins University Department of Chemistry, Zanvyl Krieger School of Arts and Sciences, and Advanced Technology Laboratory, Whiting School of Engineering, Baltimore, Maryland 21211, USA
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CellOptic, 9605 Medical Center Drive Suite 224, Rockville, Maryland 20850, USA
Joseph Rosen is on leave from the Department of Electrical and Computer Engineering, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Correspondence to: Gary Brooker1,2,3 e-mail: gbrooker@jhu.edu
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