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Rhodamine 6G conjugated to gold nanoparticles as labels for both SERS and fluorescence
studies on live endothelial cells.

Jaworska A, Wojcik T, Malek K, Kwolek U, Kepczynski M, Ansary AA, Chlopicki S, Baranska M - Mikrochim Acta (2014)

Bottom Line: The results highlight the significance of using multiple techniques to cover a spectrum of issues in the application of SERS nanosensors for probing an intracellular environment under comparable and standardized conditions.FigureCellular uptake of bare rhodamine 6G and rhodamine 6G adsorbed onto AuNPs were studied on endothelial cells using fluorescence and surface-enhanced Raman spectroscopy.The internalization of R6G-AuNPs occurs via endocytosis and diffusion resulting in uneven distribution in the cytoplasm.

View Article: PubMed Central - PubMed

Affiliation: Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland ; Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Krakow, Poland.

ABSTRACT

Fluorescence and surface-enhanced Raman scattering (SERS) spectroscopy were employed to investigate the cellular uptake of rhodamine 6G (R6G) alone and of R6G loaded with gold nanoparticles (AuNPs) by endothelial cells. R6G plays the role of a Raman reporter in SERS but also displays strong fluorescence. The presence of bare R6G molecules and R6G-AuNPs in the cytoplasm of the cells is detected via the 2D fluorescence of the dye after a 0.5 h of the incubation with R6G and R6G-AuNPs, and then the concentration of the dye increases within 4 h of exposure. The examination of the cellular uptake of the R6G and R6G-AuNPs species at different temperatures suggests that the internalization of the R6G-AuNPs into endothelial cells occurs mainly via endocytosis. 3D fluorescence imaging of R6G inside cells reveals inhomogeneous distribution of the dye in the cytoplasm. The SERS signal of the Raman reporter inside the cell disappears after 2 h of incubation with R6G-AuNPs and then amino acid residues, purines and pyrimidines become SERS-active via their interactions with the gold. The results highlight the significance of using multiple techniques to cover a spectrum of issues in the application of SERS nanosensors for probing an intracellular environment under comparable and standardized conditions. FigureCellular uptake of bare rhodamine 6G and rhodamine 6G adsorbed onto AuNPs were studied on endothelial cells using fluorescence and surface-enhanced Raman spectroscopy. The internalization of R6G-AuNPs occurs via endocytosis and diffusion resulting in uneven distribution in the cytoplasm.

No MeSH data available.


Related in: MedlinePlus

The rhodamine 6G molecules detected in cytoplasm of EA.hy 926 endothelial cells by 2D fluorescence microscopy. Nuclei stained with Hoechst 33342 are blue, R6G is yellow
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Fig2: The rhodamine 6G molecules detected in cytoplasm of EA.hy 926 endothelial cells by 2D fluorescence microscopy. Nuclei stained with Hoechst 33342 are blue, R6G is yellow

Mentions: The 2D intracellular fluorescence measurements of cells with the use of a high-content screening automated microscope enable the quantitative analysis of a dye for a large cell population [28, 29], however, this technique does not determine an exact location of the dye molecules in the intracellular compartment. Figure 2 illustrates fluorescence image of the EA.hy 926 cells with the Hoechst-stained nuclei and yellow staining of R6G distributed within cytoplasm. It is rather impossible to asses from this image whether the R6G molecules are present inside the cells, or possibly they are simply attached to the membrane surface of the endothelial cells.Fig. 2


Rhodamine 6G conjugated to gold nanoparticles as labels for both SERS and fluorescence
studies on live endothelial cells.

Jaworska A, Wojcik T, Malek K, Kwolek U, Kepczynski M, Ansary AA, Chlopicki S, Baranska M - Mikrochim Acta (2014)

The rhodamine 6G molecules detected in cytoplasm of EA.hy 926 endothelial cells by 2D fluorescence microscopy. Nuclei stained with Hoechst 33342 are blue, R6G is yellow
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4281367&req=5

Fig2: The rhodamine 6G molecules detected in cytoplasm of EA.hy 926 endothelial cells by 2D fluorescence microscopy. Nuclei stained with Hoechst 33342 are blue, R6G is yellow
Mentions: The 2D intracellular fluorescence measurements of cells with the use of a high-content screening automated microscope enable the quantitative analysis of a dye for a large cell population [28, 29], however, this technique does not determine an exact location of the dye molecules in the intracellular compartment. Figure 2 illustrates fluorescence image of the EA.hy 926 cells with the Hoechst-stained nuclei and yellow staining of R6G distributed within cytoplasm. It is rather impossible to asses from this image whether the R6G molecules are present inside the cells, or possibly they are simply attached to the membrane surface of the endothelial cells.Fig. 2

Bottom Line: The results highlight the significance of using multiple techniques to cover a spectrum of issues in the application of SERS nanosensors for probing an intracellular environment under comparable and standardized conditions.FigureCellular uptake of bare rhodamine 6G and rhodamine 6G adsorbed onto AuNPs were studied on endothelial cells using fluorescence and surface-enhanced Raman spectroscopy.The internalization of R6G-AuNPs occurs via endocytosis and diffusion resulting in uneven distribution in the cytoplasm.

View Article: PubMed Central - PubMed

Affiliation: Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland ; Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Krakow, Poland.

ABSTRACT

Fluorescence and surface-enhanced Raman scattering (SERS) spectroscopy were employed to investigate the cellular uptake of rhodamine 6G (R6G) alone and of R6G loaded with gold nanoparticles (AuNPs) by endothelial cells. R6G plays the role of a Raman reporter in SERS but also displays strong fluorescence. The presence of bare R6G molecules and R6G-AuNPs in the cytoplasm of the cells is detected via the 2D fluorescence of the dye after a 0.5 h of the incubation with R6G and R6G-AuNPs, and then the concentration of the dye increases within 4 h of exposure. The examination of the cellular uptake of the R6G and R6G-AuNPs species at different temperatures suggests that the internalization of the R6G-AuNPs into endothelial cells occurs mainly via endocytosis. 3D fluorescence imaging of R6G inside cells reveals inhomogeneous distribution of the dye in the cytoplasm. The SERS signal of the Raman reporter inside the cell disappears after 2 h of incubation with R6G-AuNPs and then amino acid residues, purines and pyrimidines become SERS-active via their interactions with the gold. The results highlight the significance of using multiple techniques to cover a spectrum of issues in the application of SERS nanosensors for probing an intracellular environment under comparable and standardized conditions. FigureCellular uptake of bare rhodamine 6G and rhodamine 6G adsorbed onto AuNPs were studied on endothelial cells using fluorescence and surface-enhanced Raman spectroscopy. The internalization of R6G-AuNPs occurs via endocytosis and diffusion resulting in uneven distribution in the cytoplasm.

No MeSH data available.


Related in: MedlinePlus