Protocol

Fluorescence Molecular Tomography of Brain Tumors in Mice

Adapted from Imaging in Neuroscience (ed. Helmchen and Konnerth). CSHL Press, Cold Spring Harbor, NY, USA, 2011.

Abstract

Fluorescence molecular tomography of tissues is a method that three-dimensionally resolves fluorescence biodistribution in vivo, with applications in small-animal research and pre-clinical diagnostics. There are many alternative imaging geometries in optical tomographic experimental systems, but in general, all imaging setups consist of four subsystems: illumination, animal mount, imaging, and automation and data acquisition (i.e., electronics and computer). Here we refer to charge-coupled device (CCD)-based systems that work in trans-illumination (i.e., illumination and detection occur on opposite sides of the subject), while a mouse or other small animal is rotated through 360° to allow photon acquisition from multiple projections. We present a procedure to tomographically reconstruct the biodistribution of fluorescence in small animals. The imaging system and equipment are described, the step-by-step image acquisition and preliminary image-processing methods are presented, and the tomographic reconstruction procedure is outlined. Finally, the method is showcased by imaging the fluorescence activity of a brain tumor of a glioblastoma mouse model.

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