Abstract
Small gold nanoparticles (sAuNPs, <10nm in a core diameter) have been used for drug delivery and cancer therapy due to their high payload to carrier ratio. Information about the amount and location of sAuNPs in cells and tissues is critical to many applications. However, the current detection method (i.e., transmission electron microscopy) for such sAuNPs is limited due to the extensive sample preparation and the limited field of view. Here we use confocal laser scanning microscopy to provide endosome-entrapped sAuNP distributions and to quantify particle uptake into cells. The quantitative capabilities of the system were confirmed by inductively coupled plasma-mass spectrometry, with an observed linear relation between scattering intensity and the initial cellular uptake of sAuNPs using 4nm and 6nm core particles.The summary of the method is:•This non-invasive imaging strategy provides a tool for label-free real-time tracking and quantification of sAuNPs using a commercially available confocal laser scanning microscope.•Scattering intensity depends on particle size.•The linear relation established between scattering intensity and uptaken gold amount enables simultaneous quantitative assessment through simple image analysis.
Highlights
Rapid, and non-invasive approach for the imaging of sAuNPs within cells by using a standard confocal laser scanning microscope (CLSM)
The sizes of AuNPs were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS) (Fig. S2)
Quantitative Imaging of endosome-entrapped sAuNP Since the cellular uptake of functionalized AuNPs is dictated by NP concentration [5], we investigated the concentration effect on the reflective imaging of intracellular AuNPs
Summary
Rapid, and non-invasive approach for the imaging of sAuNPs within cells by using a standard confocal laser scanning microscope (CLSM). Procedure Gold nanoparticles (AuNPs) were synthesized and characterized according to previous reports with slight modifications [1]. - Murray’s place-exchange method [4] was used to prepare functionalized AuNPs. The sizes of AuNPs were characterized by TEM and dynamic light scattering (DLS) (Fig. S2). - After 24 h of seeding, the cells were washed once with PBS and exposed to DMEM solution containing either 4- or 6-nm AuNPs at different concentrations (2.5, 10, 20, 40, and 60 nM for 4-nm and 0.7, 2.7, 5.5, 10.9, and 16.4 nM for 6-nm). - After 3 h incubation, the cells were washed three times with PBS to remove extra nanoparticles and used for imaging as well as ICP-MS quantification. Five confocal images were analyzed for each set of experiments
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