We report the performance of a simple method for making quantitative bioluminescence measurements of a point-like source embedded in small animals. In this method, video reflectometry is first used to obtain an estimate of the in situ optical properties of the tissue containing the bioluminescent source. A 2-dimensional image of the bioluminescence signal emitted from the surface of the animal is then acquired with a CCD. Using the measured optical properties, and a simple diffusion theory model, an inversion algorithm is applied to retrieve the source depth and power from a region of interest of the bioluminescence images. Two major factors determine the accuracy of the reconstruction: tissue heterogeneity and curvature of the imaged surface. The use of measured optical properties to characterize in situ tissue surmounts, to a degree, the heterogeneity problem: post mortem data from rats show that the relative power can be retrieved within a factor of 2 and frequently within 20 %, and the depth within 1.0 mm for implanted depths of 4-10 mm, when the curvature effects were eliminated. For depths shallower than 4 mm, the errors in the retrieved depth are consistently larger.

© 2007 SPIE

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