Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells
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 compar...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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The Royal Society
2015-01-01
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| Series: | Royal Society Open Science |
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| Online Access: | https://royalsocietypublishing.org/doi/pdf/10.1098/rsos.140454 |
| _version_ | 1818164018803638272 |
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| author | D. J. Nieves Y. Li D. G. Fernig R. Lévy |
| author_facet | D. J. Nieves Y. Li D. G. Fernig R. Lévy |
| author_sort | D. J. Nieves |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-11T16:58:47Z |
| format | Article |
| id | doaj.art-3431f73e1dd047b280ac8f0e6b760125 |
| institution | Directory Open Access Journal |
| issn | 2054-5703 |
| language | English |
| last_indexed | 2024-12-11T16:58:47Z |
| publishDate | 2015-01-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Royal Society Open Science |
| spelling | doaj.art-3431f73e1dd047b280ac8f0e6b7601252022-12-22T00:57:54ZengThe Royal SocietyRoyal Society Open Science2054-57032015-01-012610.1098/rsos.140454140454Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cellsD. J. NievesY. LiD. G. FernigR. LévyRaster 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.https://royalsocietypublishing.org/doi/pdf/10.1098/rsos.140454fluctuation spectroscopydiffusionphiricsgold nanoparticlesfgf2 |
| spellingShingle | D. J. Nieves Y. Li D. G. Fernig R. Lévy Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells Royal Society Open Science fluctuation spectroscopy diffusion phi rics gold nanoparticles fgf2 |
| title | Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells |
| title_full | Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells |
| title_fullStr | Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells |
| title_full_unstemmed | Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells |
| title_short | Photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells |
| title_sort | photothermal raster image correlation spectroscopy of gold nanoparticles in solution and on live cells |
| topic | fluctuation spectroscopy diffusion phi rics gold nanoparticles fgf2 |
| url | https://royalsocietypublishing.org/doi/pdf/10.1098/rsos.140454 |
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