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Detail of fig3D_p21_tTA025

Richi Sakaguchi, Marcus N Leiwe, Takeshi Imai (2018) Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags., eLife, Volume 7

Published in 2018 Nov 20 (Electronic publication in Nov. 20, 2018, midnight )

• doi: 10.7554/eLife.40350
• pmid: 30454553
  
• url: https://elifesciences.org/articles/40350
  

(Abstract) The stochastic multicolor labeling method 'Brainbow' is a powerful strategy to label multiple neurons differentially with fluorescent proteins; however, the fluorescence levels provided by the original attempts to use this strategy were inadequate. In the present study, we developed a stochastic multicolor labeling method with enhanced expression levels that uses a tetracycline-operator system (Tetbow). We optimized Tetbow for either plasmid or virus vector-mediated multicolor labeling. When combined with tissue clearing, Tetbow was powerful enough to visualize the three-dimensional architecture of individual neurons. Using Tetbow, we were able to visualize the axonal projection patterns of individual mitral/tufted cells along several millimeters in the mouse olfactory system. We also developed a Tetbow system with chemical tags, in which genetically encoded chemical tags were labeled with synthetic fluorophores. This was useful in expanding the repertoire of the fluorescence labels and the applications of the Tetbow system. Together, these new tools facilitate light-microscopy-based neuronal tracing at both a large scale and a high resolution.

• Animals
• Luminescent Proteins/genetics
• Luminescent Proteins/metabolism
• Mice
• Neuroanatomical Tract-Tracing Techniques/methods[major]
• Neuronal Tract-Tracers/metabolism[major]
• Olfactory Bulb/anatomy & histology
• Recombinant Proteins/genetics
• Recombinant Proteins/metabolism
• Staining and Labeling/methods[major]