Special Section on Pioneers in Biomedical Optics: A.J. Welch

Effect of optical tissue clearing on spatial resolution and sensitivity of bioluminescence imaging

[+] Author Affiliations
E. Duco Jansen, Patrick M. Pickett, Mark A. Mackanos, John Virostko

Vanderbilt University, Department of Biomedical Engineering, VU Station B #351631, Nashville, Tennessee 37235

J. Biomed. Opt. 11(4), 041119 (August 25, 2006). doi:10.1117/1.2337651
History: Received October 06, 2005; Revised February 07, 2006; Accepted February 13, 2006; Published August 25, 2006; Online August 25, 2006
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In vivo bioluminescence imaging (BLI) is a powerful method of in vivo molecular imaging based on the use of optically active luciferase reporter genes. Although this method provides superior sensitivity relative to other in vivo imaging methods, spatial resolution is poor due to light scattering. The objective of this study was to use hyperosmotic agents to reduce the scattering coefficient and hence improve spatial resolution of the BLI method. A diffusing fiber tip was used to simulate an isotropic point source of bioluminescence emission (550 to 650nm). Mouse skin was treated in vitro and in vivo with glycerol (50%, 30min) and measurements of optical properties, and imaging photon counts were made before, during, and after application of glycerol to the skin sample. Glycerol application to mouse skin had little effect on the absorption coefficient but reduced the reduced scattering coefficient by more than one order of magnitude. This effect was reversible. Consequently, the spot size (i.e., spatial resolution) of the bioluminescence point source imaged through the skin decreased by a factor of 2 (550-nm light) to 3 (650-nm light) after 30min. Simultaneously, an almost twofold decrease in the amount of light detected by the BLI system was observed, despite the fact that total transmission increased 1.7 times. We have shown here that multiply scattered light is responsible for both observations. We have shown that applying a hyperosmotic clearing agent to the skin of small rodents has the potential to improve spatial resolution of BLI owing to a reduction in the reduced scattering coefficient in the skin by one order of magnitude. However, reducing the scattering coefficient reduces the amount of light reaching the camera due to a reduction in the amount of multiply scattered light that reaches the camera aperture and thus reducing the sensitivity of the method.

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© 2006 Society of Photo-Optical Instrumentation Engineers

Citation

E. Duco Jansen ; Patrick M. Pickett ; Mark A. Mackanos and John Virostko
"Effect of optical tissue clearing on spatial resolution and sensitivity of bioluminescence imaging", J. Biomed. Opt. 11(4), 041119 (August 25, 2006). ; http://dx.doi.org/10.1117/1.2337651


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