TY - JOUR
T1 - Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
AU - Sakaguchi, Richi
AU - Leiwe, Marcus Nathanael
AU - Imai, Takeshi
N1 - Funding Information:
This work was supported by the program for Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) from Japan Agency for Medical Research and Development (AMED) (JP18dm0207055h to TI), a grant from the programs Grants-in-Aid for Scientific Research on Innovative Areas ’Dynamic regulation of Brain Function by Scrap & Build System’ (JP16H06456 to TI) from MEXT, JSPS KAKENHI (JP17H06261, JP16K14568, JP15H05572, and JP15K14336 to TI; JP17K14946 to MNL), Brain Science Foundation (to TI), and RIKEN CDB intramural grant (to TI). RS was a Junior Research Associate at RIKEN. Imaging experiments were supported by the RIKEN Kobe Light Microscopy Facility. Animal experiments were supported by LARGE. We also appreciate technical assistance from The Research Support Center, Research Center for Human Disease Modeling, Kyushu University.
Publisher Copyright:
© Sakaguchi et al.
PY - 2018/11/1
Y1 - 2018/11/1
N2 - 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.
AB - 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.
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U2 - 10.7554/eLife.40350
DO - 10.7554/eLife.40350
M3 - Article
C2 - 30454553
AN - SCOPUS:85056723503
SN - 2050-084X
VL - 7
JO - eLife
JF - eLife
M1 - e40350
ER -