Abstract
INTRODUCTIONVisualizing the precise morphology of closely juxtaposed cells and their interactions can be highly informative, particularly when studying the complex organization of neuronal and glial networks in the nervous system. To this end, one can use optical approaches to image-distinct markers that are differentially distributed among the cells of interest, such as fluorescent proteins of various colors (XFPs). The Brainbow strategies use Cre/lox recombination to stochastically express two to four XFPs in a cellular population from a single promoter. Integration of multiple Brainbow transgene copies results in combinatorial expression of these XFPs, creating a wide range of hues. In the nervous system, the multicolor labeling thus generated can be used to distinguish adjacent neuronal or glial cells and to verify the identity of neuronal processes while tracing circuitry. This article describes the generation of Brainbow transgenes and mice as well as their use to image and digitally reconstruct nerve cells and their interactions in fixed samples. This method also holds potential for studies in other tissues and model organisms as well as live imaging in vivo.
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