Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags

Richi Sakaguchi, Marcus N. Leiwe, Takeshi Imai

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish
Article numbere40350
JournaleLife
Volume7
DOIs
Publication statusPublished - Nov 1 2018

Fingerprint

Labeling
Fluorescence
Neurons
Networks (circuits)
Labels
Tetracycline
Microscopy
Proteins
Plasmids
Fluorophores
Viruses
Light
Optical microscopy
Tissue

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags. / Sakaguchi, Richi; Leiwe, Marcus N.; Imai, Takeshi.

In: eLife, Vol. 7, e40350, 01.11.2018.

Research output: Contribution to journalArticle

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