Super-photostable organic dye for long-term live-cell single-protein imaging

Do-Hyeon Kim; Hong Minh Triet; Sun Hyeok Lee; Sina Jazani; Seongjae Jang; Syed Ali Abbas Abedi; Xiaogang Liu; Jongcheol Seo; Taekjip Ha; Young-Tae Chang; Sung Ho Ryu

doi:10.1038/s41592-024-02584-0

Super-photostable organic dye for long-term live-cell single-protein imaging

Nat Methods. 2025 Jan 15. doi: 10.1038/s41592-024-02584-0. Online ahead of print.

Authors

Do-Hyeon Kim^#^{1

2}, Hong Minh Triet^#^{3

4

5}, Sun Hyeok Lee^#⁶, Sina Jazani⁷, Seongjae Jang⁴, Syed Ali Abbas Abedi⁸, Xiaogang Liu⁸, Jongcheol Seo^{4

5

6}, Taekjip Ha^{7

9

10

11}, Young-Tae Chang^{12

13

14}, Sung Ho Ryu¹⁵

Affiliations

¹ Department of Life Sciences, Pohang University of Science and Technology, Pohang, Republic of Korea. [email protected].
² Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Maryland, Baltimore, MD, USA. [email protected].
³ Department of Life Sciences, Pohang University of Science and Technology, Pohang, Republic of Korea.
⁴ Department of Chemistry, Pohang University of Science and Technology, Pohang, Republic of Korea.
⁵ Molecular Imaging Center, Pohang University of Science and Technology, Pohang, Republic of Korea.
⁶ School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Republic of Korea.
⁷ Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Maryland, Baltimore, MD, USA.
⁸ Science, Mathematics and Technology Cluster, Singapore University of Technology and Design, Singapore, Singapore.
⁹ Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.
¹⁰ Department of Pediatrics, Harvard Medical School, Boston, MA, USA.
¹¹ Howard Hughes Medical Institute, Boston, MA, USA.
¹² Department of Chemistry, Pohang University of Science and Technology, Pohang, Republic of Korea. [email protected].
¹³ Molecular Imaging Center, Pohang University of Science and Technology, Pohang, Republic of Korea. [email protected].
¹⁴ School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Republic of Korea. [email protected].
¹⁵ Department of Life Sciences, Pohang University of Science and Technology, Pohang, Republic of Korea. [email protected].

^# Contributed equally.

PMID: 39815105
DOI: 10.1038/s41592-024-02584-0

Abstract

Organic dyes play a crucial role in live-cell imaging because of their advantageous properties, such as photostability and high brightness. Here we introduce a super-photostable and bright organic dye, Phoenix Fluor 555 (PF555), which exhibits an order-of-magnitude longer photobleaching lifetime than conventional organic dyes without the requirement of any anti-photobleaching additives. PF555 is an asymmetric cyanine structure in which, on one side, the indole in the conventional Cyanine-3 is substituted with 3-oxo-quinoline. PF555 provides a powerful tool for long-term live-cell single-molecule imaging, as demonstrated by the imaging of the dynamic single-molecule interactions of the epidermal growth factor receptor with clathrin-coated structures on the plasma membrane of a live cell under physiological conditions.