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Bioconjug Chem. 2018 Apr 18;29(4):1327-1334. doi: 10.1021/acs.bioconjchem.8b00069. Epub 2018 Feb 26.

Autofluorescence-Free Live-Cell Imaging Using Terbium Nanoparticles.

Author information

1
NanoBioPhotonics (nanofret.com), Institute for Integrative Biology of the Cell (I2BC) , Université Paris-Saclay, Univeristé Paris-Sud, CNRS, CEA , 91405 Orsay Cedex, France.
2
Laboratoire d'Ingénierie Moléculaire Appliquée à l'Analyse (LIMAA) , Institut Pluridisciplinaire Hubert Curien (IPHC), CNRS, Université de Strasbourg , 67087 Strasbourg Cedex, France.
3
Department of Chemistry , Hong Kong Baptist University , Hong Kong SAR , Hong Kong.

Abstract

Fluorescent nanoparticles (NPs) have become irreplaceable tools for advanced cellular and subcellular imaging. While very bright NPs require excitation with UV or visible light, which can create strong autofluorescence of biological components, NIR-excitable NPs without autofluorescence issues exhibit much lower brightness. Here, we show the application of a new type of surface-photosensitized terbium NPs (Tb-NPs) for autofluorescence-free intracellular imaging in live HeLa cells. The combination of exceptionally high brightness, high photostability, and long photoluminecence (PL) lifetimes for highly efficient suppression of the short-lived autofluorescence allowed for time-gated PL imaging of intracellular vesicles over 72 h without toxicity and at extremely low Tb-NP concentrations down to 12 pM. Detection of highly resolved long-lifetime (ms) PL decay curves from small (∼10 μm2) areas within single cells within a few seconds emphasized the unprecedented photophysical properties of Tb-NPs for live-cell imaging that extend well beyond currently available nanometric imaging agents.

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