Optical projection tomography as a tool for 3D microscopy and gene expression studies

Current techniques for three-dimensional (3D) optical microscopy (deconvolution, confocal. microscopy, and optical coherence tomography) generate 3D data by optically sectioning the specimen. This places severe constraints on the maximum thickness of a specimen that can be imaged. We have developed a microscopy technique that uses optical projection tomography (OPT) to produce high-resolution 3D images of both fluorescent and nonfluorescent biological specimens with a thickness of up to 15 millimeters. OPT microscopy allows the rapid mapping of the tissue distribution of RNA and protein expression in intact embryos or organ systems and can therefore be instrumental in studies of developmental biology or gene function.