Fluorescence life-time imaging and steady state polarization for examining binding of fluorophores to gold nanoparticles

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17 Scopus citations

Abstract

Nanocomposites as multifunctional agents are capable of combing imaging and cell biology technologies. The conventional methods used for validation of the conjugation process of nanoparticles (NPs) to fluorescent molecules such as spectroscopy analysis and surface potential measurements, are not sufficient. In this paper we present a new and highly sensitive procedure that uses the combination of (1) fluorescence spectrum, (2) fluorescence lifetime, and (3) steady state fluorescence polarization measurements. We characterize and analyze gold NPs with Lucifer yellow (LY) surface coating as a model. We demonstrate the ability to differentiate between LY-GNP (the conjugated complex) and a mixture of coated NP and free dyes. We suggest the approach for neuroscience applications where LY is used for detecting and labeling cells, studying morphology and intracellular communications. Histograms of Fluorescence lifetime imaging (FLIM) of free LY dye (Left) in comparison to the conjugated dye to gold nanoparticles, LY-GNP (Middle) enable the differentiation between LY-GNP (the conjugated complex) and a mixture of coated NP and free dyes (Right). Nanoparticles are commonly used for imaging and cell biology technologies. One of the most popular nanoparticle-based complexes is the conjugation of nanoparticles to fluorescent molecules. The conventional methods used for validation of the conjugation process such as spectroscopy analysis and surface potential measurements, are not sufficient. In this paper we present a new approach that uses the combination of (1) fluorescence spectrum, (2) fluorescence lifetime, and (3) steady state fluorescence polarization measurements, to differentiate between samples of nanoparticles coated with Lucifer yellow and samples of the free dye.

Original languageEnglish
Pages (from-to)944-951
Number of pages8
JournalJournal of Biophotonics
Volume8
Issue number11-12
DOIs
StatePublished - Nov 2015

Bibliographical note

Publisher Copyright:
© 2015 WILEY-VCH Verlag GmbH & Co.

Keywords

  • Fluorescence life time
  • Fluorescence polarization
  • Gold nanoparticles
  • Nanoparticles

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