Research Papers

Design and quantitative resolution measurements of an optical virtual sectioning three-dimensional imaging technique for biomedical specimens, featuring two-micrometer slicing resolution

[+] Author Affiliations
Jan A. N. Buytaert, Joris J. J. Dirckx

University of Antwerp, Laboratory of Biomedical Physics, Department of Physics, Groenenborgerlaan 171, B-2020 Wilrijk-Antwerpen, Belgium

J. Biomed. Opt. 12(1), 014039 (February 23, 2007). doi:10.1117/1.2671712
History: Received March 31, 2006; Revised August 25, 2006; Accepted August 25, 2006; Published February 23, 2007
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Several well-established techniques are available to obtain 3-D image information of biomedical specimens, each with their specific advantages and limitations. Orthogonal plane fluorescence optical sectioning (OPFOS), or selective plane illumination microscopy (SPIM), are additional techniques which, after adequate specimen preparation, produce high quality, autoaligned sectional images in nearly real time, of bone as well as soft tissue. Up until now, slicing resolutions down to 14μm have been obtained. We present a high resolution (HR) OPFOS method, which delivers images that approach the quality of histological sections. With our HROPFOS technique, we achieve in-plane resolutions of 1μm and a slicing resolution of 2μm. A region of interest within an intact and much larger object can be imaged without problems, and as the optical technique is nondestructive, the object can be measured in any slicing direction. We present quantitative measurements of resolution. A 3-D model reconstructed from our HROPFOS data is compared to SEM results, and the technique is demonstrated with section images and 3-D reconstructions of middle ear specimens.

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© 2007 Society of Photo-Optical Instrumentation Engineers

Citation

Jan A. N. Buytaert and Joris J. J. Dirckx
"Design and quantitative resolution measurements of an optical virtual sectioning three-dimensional imaging technique for biomedical specimens, featuring two-micrometer slicing resolution", J. Biomed. Opt. 12(1), 014039 (February 23, 2007). ; http://dx.doi.org/10.1117/1.2671712


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