Nature 450, 56-62 (1 November 2007) | doi:10.1038/nature06293; Received 17 July 2007; Accepted 25 September 2007

Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system

Jean Livet1, Tamily A. Weissman1, Hyuno Kang1, Ryan W. Draft1, Ju Lu1, Robyn A. Bennis1, Joshua R. Sanes1 & Jeff W. Lichtman1

  1. Department of Molecular and Cellular Biology and Center for Brain Science, Harvard University, Cambridge, Massachusetts 02138, USA

Correspondence to: Jeff W. Lichtman1 Correspondence and requests for materials should be addressed to J.W.L. (Email:


Detailed analysis of neuronal network architecture requires the development of new methods. Here we present strategies to visualize synaptic circuits by genetically labelling neurons with multiple, distinct colours. In Brainbow transgenes, Cre/lox recombination is used to create a stochastic choice of expression between three or more fluorescent proteins (XFPs). Integration of tandem Brainbow copies in transgenic mice yielded combinatorial XFP expression, and thus many colours, thereby providing a way to distinguish adjacent neurons and visualize other cellular interactions. As a demonstration, we reconstructed hundreds of neighbouring axons and multiple synaptic contacts in one small volume of a cerebellar lobe exhibiting approximately 90 colours. The expression in some lines also allowed us to map glial territories and follow glial cells and neurons over time in vivo. The ability of the Brainbow system to label uniquely many individual cells within a population may facilitate the analysis of neuronal circuitry on a large scale.


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