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Amphipathic Fluorescent Dyes for Sensitive and Long-Term Monitoring of Plasma Membranes

Masataka Takahashi, Ryo Seino, Takatoshi Ezoe, Munetaka Ishiyama, Yuichiro Ueno
doi: https://doi.org/10.1101/2020.11.16.379206
Masataka Takahashi
Dojindo Laboratories, Tabaru 2025-5, Mashiki-machi, Kumamoto 861-2202, Japan
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  • For correspondence: m.takahashi@dojindo.co.jp
Ryo Seino
Dojindo Laboratories, Tabaru 2025-5, Mashiki-machi, Kumamoto 861-2202, Japan
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Takatoshi Ezoe
Dojindo Laboratories, Tabaru 2025-5, Mashiki-machi, Kumamoto 861-2202, Japan
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Munetaka Ishiyama
Dojindo Laboratories, Tabaru 2025-5, Mashiki-machi, Kumamoto 861-2202, Japan
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Yuichiro Ueno
Dojindo Laboratories, Tabaru 2025-5, Mashiki-machi, Kumamoto 861-2202, Japan
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ABSTRACT

The plasma membrane (PM) plays a critical role in many cellular processes, and PM dysfunction is a key biomarker related to the cell status and several diseases. Imaging techniques using small fluorescent probes have become increasingly important tools for visualizing living cells, particularly their PMs. Among the commercially available PM-specific probes, PKH dyes are widely used; however, the utility of these dyes is limited by their short membrane retention times and high cytotoxicity. Herein, PlasMem Bright Green and Red are implemented as new PM-specific fluorescent probes, which employ a polycyclic aromatic fluorophore to improve their retention ability and a strong acid moiety to reduce their transmembrane diffusion and cytotoxicity. We demonstrate that the long retention and low cytotoxicity of the PlasMem Bright dyes enable them to be applied for observing neuronal PMs and monitoring PM dynamics involving the endocytic pathway. Furthermore, we successfully detected mitochondria in nerve axons over long periods using PlasMem Bright dyes. Finally, the combined use of exosome staining probes and PlasMem Bright dyes allowed clear visualization of the endocytic pathway.

Figure1

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • The authors declare no competing financial interests.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 17, 2020.
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Amphipathic Fluorescent Dyes for Sensitive and Long-Term Monitoring of Plasma Membranes
Masataka Takahashi, Ryo Seino, Takatoshi Ezoe, Munetaka Ishiyama, Yuichiro Ueno
bioRxiv 2020.11.16.379206; doi: https://doi.org/10.1101/2020.11.16.379206
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Amphipathic Fluorescent Dyes for Sensitive and Long-Term Monitoring of Plasma Membranes
Masataka Takahashi, Ryo Seino, Takatoshi Ezoe, Munetaka Ishiyama, Yuichiro Ueno
bioRxiv 2020.11.16.379206; doi: https://doi.org/10.1101/2020.11.16.379206

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