Uptake study of gold nanoparticles into cancer cells using high-order image correlation spectroscopy

2019 ◽  
Author(s):  
D. Katoozi ◽  
A. H. A. Clayton ◽  
D. J. Moss ◽  
J. W. M. Chon
Keyword(s):  
Gold Nanoparticles ◽  
Cancer Cells ◽  
High Order ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Uptake Study ◽  
Image Correlation Spectroscopy ◽  
Order Image

scholarly journals Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells

2015 ◽  
Vol 2 (6) ◽  
pp. 140454 ◽  
Author(s):  
D. J. Nieves ◽  
Y. Li ◽  
D. G. Fernig ◽  
R. Lévy
Keyword(s):  
Gold Nanoparticles ◽  
Single Molecule ◽  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Live Cells ◽  
Glycerol Water ◽  
Raster Image ◽  
Diffusion Speed ◽  
Image Correlation Spectroscopy ◽  
Representative Area

Raster image correlation spectroscopy (RICS) measures the diffusion of fluorescently labelled molecules from stacks of confocal microscopy images by analysing correlations within the image. RICS enables the observation of a greater and, thus, more representative area of a biological system as compared to other single molecule approaches. Photothermal microscopy of gold nanoparticles allows long-term imaging of the same labelled molecules without photobleaching. Here, we implement RICS analysis on a photothermal microscope. The imaging of single gold nanoparticles at pixel dwell times short enough for RICS (60 μs) with a piezo-driven photothermal heterodyne microscope is demonstrated (photothermal raster image correlation spectroscopy, PhRICS). As a proof of principle, PhRICS is used to measure the diffusion coefficient of gold nanoparticles in glycerol : water solutions. The diffusion coefficients of the nanoparticles measured by PhRICS are consistent with their size, determined by transmission electron microscopy. PhRICS was then used to probe the diffusion speed of gold nanoparticle-labelled fibroblast growth factor 2 (FGF2) bound to heparan sulfate in the pericellular matrix of live fibroblast cells. The data are consistent with previous single nanoparticle tracking studies of the diffusion of FGF2 on these cells. Importantly, the data reveal faster FGF2 movement, previously inaccessible by photothermal tracking, and suggest that inhomogeneity in the distribution of bound FGF2 is dynamic.

Exploring oligomeric state of the serotonin1A receptor utilizing photobleaching image correlation spectroscopy: implications for receptor function

2018 ◽  
Vol 207 ◽  
pp. 409-421 ◽  
Author(s):  
Hirak Chakraborty ◽  
Md. Jafurulla ◽  
Andrew H. A. Clayton ◽  
Amitabha Chattopadhyay
Keyword(s):  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Membrane Cholesterol ◽  
Receptor Function ◽  
Oligomeric State ◽  
Serotonin1a Receptor ◽  
Image Correlation Spectroscopy

Photobleaching image correlation spectroscopy (pbICS) reveals that membrane cholesterol modulates the oligomeric state of the serotonin1A receptor.

scholarly journals Arbitrary-Region Image Correlation Spectroscopy

2016 ◽  
Vol 110 (3) ◽  
pp. 176a
Author(s):  
Jelle Hendrix ◽  
Tomas Dekens ◽  
Don C. Lamb
Keyword(s):  
Correlation Spectroscopy ◽  
Image Correlation ◽  
Arbitrary Region ◽  
Image Correlation Spectroscopy

scholarly journals Protein-Coated Aryl Modified Gold Nanoparticles for Cellular Uptake Study by Osteosarcoma Cancer Cells

2020 ◽  
Author(s):  
Mehvesh Hameed ◽  
Seema Panicker ◽  
Sallam Hasan Abdallah ◽  
Amir A. Khan ◽  
Changseok Han ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Cancer Cells ◽  
Cellular Uptake ◽  
Phase Transfer ◽  
Chemical Reduction ◽  
Uptake Study ◽  
Stabilizing Agents ◽  
Coated Nanoparticles ◽  
Green Route ◽  
The Difference

We synthesized protein-coated gold nanoparticles using green and chemical reduction routes for cellular uptake study. In the current work, we coated gold-aryl nanoparticles of the type AuNPs-C<sub>6</sub>H<sub>4</sub>-4-COOH with BSA, collagen, zein and lysozyme proteins. Both routes were carried out without phase-transfer catalysts or extraneous stabilizing agents. High crystallinity of the AuNPs synthesized by the green route can be seen in the transmission electron microscopy images. <a>Osteosarcoma cancer cells are malignant bone tumors with abnormal cellular functions. Studies using MG-63 cells will provide mechanistic suggestions on the details of the amplification in tumors. </a>We studied the cellular uptake of the bioconjugates by MG-63 osteosarcoma cells using laser confocal fluorescence microscopy (LCFM) and flow cytometry. In the LCFM study, BSA-AuNPs was uptaken most efficiently of all protein-coated gold nanoparticles synthesized by the green route. Zein and lysozyme coated nanoparticles, though small sizes, prepared by the green method were not efficiently uptaken by MG-63. The two nanoparticles are negatively charged and zein is also a hydrophobic coat. The difference in hydrophobicity and charge might have affected the internalization. All of those coated nanoparticles that were efficiently uptaken can potentially be used as diagnostic and therapeutic agents for osteosarcoma.

scholarly journals Visualization of class A GPCR oligomerization by image-based fluorescence fluctuation spectroscopy

2017 ◽  
Author(s):  
Ali Isbilir ◽  
Jan Möller ◽  
Andreas Bock ◽  
Ulrike Zabel ◽  
Paolo Annibale ◽  
...  
Keyword(s):  
Single Molecule ◽  
Metabotropic Glutamate Receptors ◽  
Fluorescent Protein ◽  
Methodological Approach ◽  
Correlation Spectroscopy ◽  
Membrane Receptors ◽  
Image Correlation ◽  
Intact Cells ◽  
Class A ◽  
Image Correlation Spectroscopy

AbstractG protein-coupled receptors (GPCRs) represent the largest class of cell surface receptors conveying extracellular information into intracellular signals. Many GPCRs have been shown to be able to oligomerize and it is firmly established that Class C GPCRs (e.g. metabotropic glutamate receptors) function as obligate dimers. However, the oligomerization capability of the larger Class A GPCRs (e.g. comprising the β-adrenergic receptors (β-ARs)) is still, despite decades of research, highly debated.Here we assess the oligomerization behavior of three prototypical Class A GPCRs, the β1-ARs, β2-ARs, and muscarinic M2Rs in single, intact cells. We combine two image correlation spectroscopy methods based on molecular brightness, i.e. the analysis of fluorescence fluctuations over space and over time, and thereby provide an assay able to robustly and precisely quantify the degree of oligomerization of GPCRs. In addition, we provide a comparison between two labelling strategies, namely C-terminally-attached fluorescent proteins and N-terminally-attached SNAP-tags, in order to rule out effects arising from potential fluorescent protein-driven oligomerization. The degree of GPCR oligomerization is expressed with respect to a set of previously reported as well as newly established monomeric or dimeric control constructs. Our data reveal that all three prototypical GPRCs studied display, under unstimulated conditions, a prevalently monomeric fingerprint. Only the β2-AR shows a slight degree of oligomerization.From a methodological point of view, our study suggests three key aspects. First, the combination of two image correlation spectroscopy methods allows addressing cells transiently expressing high concentrations of membrane receptors, far from the single molecule regime, at a density where the kinetic equilibrium should favor dimers and higher-order oligomers. Second, our methodological approach, allows to selectively target cell membrane regions devoid of artificial oligomerization hot-spots (such as vesicles). Third, our data suggest that the β1-AR appears to be a superior monomeric control than the widely used membrane protein CD86.Taken together, we suggest that our combined image correlation spectroscopy method is a powerful approach to assess the oligomerization behavior of GPCRs in intact cells at high expression levels.

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