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J Am Chem Soc. 2010 Apr 14;132(14):5021-3. doi: 10.1021/ja100079w.

Spectroscopic rationale for efficient stimulated-emission depletion microscopy fluorophores.

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  • 1Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, B-3001 Heverlee, Belgium. junichi.hotta@chem.kuleuven.be

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.

PMID:
20307062
[PubMed - indexed for MEDLINE]
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