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Journal of Biomedical Optics • Open Access

Optically sectioned in vivo imaging with speckle illumination HiLo microscopy
Author(s): Daryl Lim; Timothy N. Ford; Kengyeh K. Chu; Jerome Metz

Paper Abstract

We present a simple wide-field imaging technique, called HiLo microscopy, that is capable of producing optically sectioned images in real time, comparable in quality to confocal laser scanning microscopy. The technique is based on the fusion of two raw images, one acquired with speckle illumination and another with standard uniform illumination. The fusion can be numerically adjusted, using a single parameter, to produce optically sectioned images of varying thicknesses with the same raw data. Direct comparison between our HiLo microscope and a commercial confocal laser scanning microscope is made on the basis of sectioning strength and imaging performance. Specifically, we show that HiLo and confocal 3-D imaging of a GFP-labeled mouse brain hippocampus are comparable in quality. Moreover, HiLo microscopy is capable of faster, near video rate imaging over larger fields of view than attainable with standard confocal microscopes. The goal of this paper is to advertise the simplicity, robustness, and versatility of HiLo microscopy, which we highlight with in vivo imaging of common model organisms including planaria, C. elegans, and zebrafish.

Paper Details

Date Published: 1 January 2011
PDF: 8 pages
J. Biomed. Opt. 16(1) 016014 doi: 10.1117/1.3528656
Published in: Journal of Biomedical Optics Volume 16, Issue 1
Show Author Affiliations
Daryl Lim, Boston Univ. (United States)
Timothy N. Ford, Boston Univ. (United States)
Kengyeh K. Chu, Boston Univ. (United States)
Jerome Metz, Boston Univ. (United States)

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