Work

Spectroscopic Rationale for Efficient Stimulated-Emission Depletion Microscopy Fluorophores

Year: 2010

Type: article

Abstract: We report a rationale for identifying superior dyes for stimulated-emission depletion (STED) microscopy. We compared the dyes pPDI and pTDI, which displayed excellent photostability in single-molecule spectroscopy. Surprisingly, their photostability and performance in STED microscopy differed significantly. While single pTDI molecules could be visualized with excellent resolution (35 nm), pPDI molecules bleached rapidly under similar conditions. Femtosecond transient absorption measurements proved that the overlap between the stimulated-emission band and the excited-state absorption band is the main reason for the observed difference. Thus, assessment of the excited-state absorption band provides a rational means of dye selection and determination of the optimal wavelength for STED.

Source: Journal of the American Chemical Society

DOI: 10.1021/ja100079w

Authors: Jun-ichi Hotta, Eduard Fron, Peter Dedecker, Kris P. F. Janssen, Chen Li, Klaus Müllen, Benjamin Harke, Johanna Bückers, Stefan W. Hell, Johan Hofkens

Institutions: Max Planck Institute for Polymer Research, Max Planck Institute for Biophysical Chemistry, KU Leuven

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: European Science Foundation, Fonds Wetenschappelijk Onderzoek, Vlaamse regering

OpenAlex ID: W1967640331

FWCI: 5.4

Cites: 31

Cited by: 109

Related to: 10