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Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
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 mul...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6245733/ https://www.ncbi.nlm.nih.gov/pubmed/30454553 http://dx.doi.org/10.7554/eLife.40350 |
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author | Sakaguchi, Richi Leiwe, Marcus N Imai, Takeshi |
author_facet | Sakaguchi, Richi Leiwe, Marcus N Imai, Takeshi |
author_sort | Sakaguchi, Richi |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-6245733 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-62457332018-11-20 Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags Sakaguchi, Richi Leiwe, Marcus N Imai, Takeshi eLife Neuroscience 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. eLife Sciences Publications, Ltd 2018-11-20 /pmc/articles/PMC6245733/ /pubmed/30454553 http://dx.doi.org/10.7554/eLife.40350 Text en © 2018, Sakaguchi et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Neuroscience Sakaguchi, Richi Leiwe, Marcus N Imai, Takeshi Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags |
title | Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags |
title_full | Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags |
title_fullStr | Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags |
title_full_unstemmed | Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags |
title_short | Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags |
title_sort | bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6245733/ https://www.ncbi.nlm.nih.gov/pubmed/30454553 http://dx.doi.org/10.7554/eLife.40350 |
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