ABSTRACT
The response of water-soluble, mercaptocarboxylic acid-capped fluorescent semiconductor nanoparticles, or quantum dots (QDs), to extended visible-light irradiation is variable and poorly described. Here we use time-resolved spectroscopy to investigate the photoluminescence intensities and lifetimes of CdSe/ZnS and CdTe QDs as a function of blue light illumination. Conjugates of the particles to the electron donor dopamine were also investigated, and the effect of the antioxidant beta-mercaptoethanol was explored. Both types of QD showed signs of direct electron transfer to the conjugate, but enhancement was much more pronounced in CdSe/ZnS. A model of the two different types of enhancement is proposed.
Subject(s)
Cadmium Compounds/chemistry , Dopamine/chemistry , Quantum Dots , Selenium Compounds/chemistry , Sulfides/chemistry , Tellurium/chemistry , Zinc Compounds/chemistry , Luminescence , Oxygen/chemistry , Photons , Time FactorsABSTRACT
Chemical modification of the surface of CdSe/ZnS quantum dots (QDs) with small molecules or functional ligands often alters the characteristics of these particles. For instance, dopamine conjugation quenches the fluorescence of the QDs, which is a property that can be exploited for sensing applications if the conjugates are taken up into living cells. However, different sizes and/or preparations of mercaptocarboxylic acid solubilized QDs show very different properties when incubated with cells. It is unknown what physical parameters determine a QDs ability to interact with a cell surface, be endocytosed, escape from endosomes, and/or enter the nucleus. In this study, we examine the surface chemistry of QD-dopamine conjugates and present an optimized method for tracking the attachment of small biomolecules to the surface. It is found that the fluorescence intensity, surface charge, colloidal stability, and biological interactions of the QDs vary as a function of the density of dopamine on the surface. Successful targeting of QD-dopamine to dopamine receptor positive PC12 cells correlates with greater homogeneity of particle thiol layer, and a minimum number of ligands required for specific association can be estimated. These results will enable users to develop methods for screening QD conjugates for biological activity before proceeding to experiments with cell lines and animals.
Subject(s)
Cadmium Compounds/chemistry , Quantum Dots , Selenium Compounds/chemistry , Sulfides/chemistry , Zinc Compounds/chemistry , Animals , Electrophoresis, Agar Gel , Ligands , PC12 Cells , Rats , Spectrum Analysis/methods , Surface PropertiesABSTRACT
CdSe/ZnS quantum dots (QDs) conjugated to biomolecules that quench their fluorescence, particularly dopamine, have particular spectral properties that allow determination of the number of conjugates per particle, namely, photoenhancement and photobleaching. In this work, we quantify these properties on a single-particle and ensemble basis in order to evaluate their usefulness as a tool for indicating QD uptake, breakdown, and processing in living cells. This creates a general framework for the use of fluorescence quenching and intermittency to better understand nanoparticle-cell interactions.
Subject(s)
Dopamine/metabolism , Fluorescence Recovery After Photobleaching/methods , Neurons/metabolism , Quantum Dots , Spectrometry, Fluorescence/methods , Algorithms , Animals , HumansABSTRACT
Semiconductor quantum dots (QDs) have been widely used for fluorescent labelling. However, their ability to transfer electrons and holes to biomolecules leads to spectral changes and effects on living systems that have yet to be exploited. Here we report the first cell-based biosensor based on electron transfer between a small molecule (the neurotransmitter dopamine) and CdSe/ZnS QDs. QD-dopamine conjugates label living cells in a redox-sensitive pattern: under reducing conditions, fluorescence is only seen in the cell periphery and lysosomes. As the cell becomes more oxidizing, QD labelling appears in the perinuclear region, including in or on mitochondria. With the most-oxidizing cellular conditions, QD labelling throughout the cell is seen. Phototoxicity results from the creation of singlet oxygen, and can be reduced with antioxidants. This work suggests methods for the creation of phototoxic drugs and for redox-specific fluorescent labelling that are generalizable to any QD conjugated to an electron donor.
