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Analysis of intracellular state based on controlled 3D nanostructures mediated surface enhanced Raman scattering.

El-Said WA, Kim TH, Kim H, Choi JW - PLoS ONE (2011)

Bottom Line: This SERS-active surface was applied as cell culture system to study living cells in situ within their culture environment without any external preparation processes.We applied this newly developed method to cell-based research to differentiate cell lines, cells at different cell cycle stages, and live/dead cells.The enhanced Raman signals achieved from each cell, which represent the changes in biochemical compositions, enabled differentiation of each state and the conditions of the cells.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Program of Integrated Biotechnology, Sogang University, Seoul, Republic of Korea.

ABSTRACT
Near-infrared surface-enhanced Raman spectroscopy (SERS) is a powerful technique for analyzing the chemical composition within a single living cell at unprecedented resolution. However, current SERS methods employing uncontrollable colloidal metal particles or non-uniformly distributed metal particles on a substrate as SERS-active sites show relatively low reliability and reproducibility. Here, we report a highly-ordered SERS-active surface that is provided by a gold nano-dots array based on thermal evaporation of gold onto an ITO surface through a nanoporous alumina mask. This new combined technique showed a broader distribution of hot spots and a higher signal-to-noise ratio than current SERS techniques due to the highly reproducible and uniform geometrical structures over a large area. This SERS-active surface was applied as cell culture system to study living cells in situ within their culture environment without any external preparation processes. We applied this newly developed method to cell-based research to differentiate cell lines, cells at different cell cycle stages, and live/dead cells. The enhanced Raman signals achieved from each cell, which represent the changes in biochemical compositions, enabled differentiation of each state and the conditions of the cells. This SERS technique employing a tightly controlled nanostructure array can potentially be applied to single cell analysis, early cancer diagnosis and cell physiology research.

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Related in: MedlinePlus

SERS mapping image and spectra of HEK 293T cells.(A) SERS map image of cells incubated with colloidal Au NPs. (B) SERS spectra from twenty different points inside the cell nucleus incubated with colloidal Au NPs. (C) Mean SERS spectrum of twenty spectra at different points inside the cell nucleus incubated with colloidal Au NPs. (D) SERS map image of cells immobilized on the randomly distributed Au NP array. (E) SERS spectra from twenty different points inside the cell nucleus immobilized on the Au NP array. (F) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au NP array. (G) SERS map image of cells immobilized on the homogenous Au nanodot array. (H) SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. (I) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. Scale bar: 10 µm.
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pone-0015836-g003: SERS mapping image and spectra of HEK 293T cells.(A) SERS map image of cells incubated with colloidal Au NPs. (B) SERS spectra from twenty different points inside the cell nucleus incubated with colloidal Au NPs. (C) Mean SERS spectrum of twenty spectra at different points inside the cell nucleus incubated with colloidal Au NPs. (D) SERS map image of cells immobilized on the randomly distributed Au NP array. (E) SERS spectra from twenty different points inside the cell nucleus immobilized on the Au NP array. (F) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au NP array. (G) SERS map image of cells immobilized on the homogenous Au nanodot array. (H) SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. (I) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. Scale bar: 10 µm.

Mentions: SERS mapping was then conducted for each cell (Figures 3A, 3D, and 3G), which enabled the SERS spectra to be obtained at each point within the map images. As shown in Figure 3A, the SERS map image of cells incubated with the colloidal Au NPs solution revealed unclear or no cell image due to the limited delivery of Au NPs into cells. However, the SERS map image of cells immobilized on a non–homogeneous Au NPs substrate produced a better image than that of the colloidal Au NP solution (Figure 3D). Conversely, cells on the Au nano–dot substrate (Figure 3G) yielded a high resolution SERS map image, which clearly differentiated the cytoplasm and nucleus of the cells.


Analysis of intracellular state based on controlled 3D nanostructures mediated surface enhanced Raman scattering.

El-Said WA, Kim TH, Kim H, Choi JW - PLoS ONE (2011)

SERS mapping image and spectra of HEK 293T cells.(A) SERS map image of cells incubated with colloidal Au NPs. (B) SERS spectra from twenty different points inside the cell nucleus incubated with colloidal Au NPs. (C) Mean SERS spectrum of twenty spectra at different points inside the cell nucleus incubated with colloidal Au NPs. (D) SERS map image of cells immobilized on the randomly distributed Au NP array. (E) SERS spectra from twenty different points inside the cell nucleus immobilized on the Au NP array. (F) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au NP array. (G) SERS map image of cells immobilized on the homogenous Au nanodot array. (H) SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. (I) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. Scale bar: 10 µm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3044723&req=5

pone-0015836-g003: SERS mapping image and spectra of HEK 293T cells.(A) SERS map image of cells incubated with colloidal Au NPs. (B) SERS spectra from twenty different points inside the cell nucleus incubated with colloidal Au NPs. (C) Mean SERS spectrum of twenty spectra at different points inside the cell nucleus incubated with colloidal Au NPs. (D) SERS map image of cells immobilized on the randomly distributed Au NP array. (E) SERS spectra from twenty different points inside the cell nucleus immobilized on the Au NP array. (F) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au NP array. (G) SERS map image of cells immobilized on the homogenous Au nanodot array. (H) SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. (I) Mean SERS spectra of twenty different points inside the cell nucleus immobilized on the Au nano-dot array. Scale bar: 10 µm.
Mentions: SERS mapping was then conducted for each cell (Figures 3A, 3D, and 3G), which enabled the SERS spectra to be obtained at each point within the map images. As shown in Figure 3A, the SERS map image of cells incubated with the colloidal Au NPs solution revealed unclear or no cell image due to the limited delivery of Au NPs into cells. However, the SERS map image of cells immobilized on a non–homogeneous Au NPs substrate produced a better image than that of the colloidal Au NP solution (Figure 3D). Conversely, cells on the Au nano–dot substrate (Figure 3G) yielded a high resolution SERS map image, which clearly differentiated the cytoplasm and nucleus of the cells.

Bottom Line: This SERS-active surface was applied as cell culture system to study living cells in situ within their culture environment without any external preparation processes.We applied this newly developed method to cell-based research to differentiate cell lines, cells at different cell cycle stages, and live/dead cells.The enhanced Raman signals achieved from each cell, which represent the changes in biochemical compositions, enabled differentiation of each state and the conditions of the cells.

View Article: PubMed Central - PubMed

Affiliation: Interdisciplinary Program of Integrated Biotechnology, Sogang University, Seoul, Republic of Korea.

ABSTRACT
Near-infrared surface-enhanced Raman spectroscopy (SERS) is a powerful technique for analyzing the chemical composition within a single living cell at unprecedented resolution. However, current SERS methods employing uncontrollable colloidal metal particles or non-uniformly distributed metal particles on a substrate as SERS-active sites show relatively low reliability and reproducibility. Here, we report a highly-ordered SERS-active surface that is provided by a gold nano-dots array based on thermal evaporation of gold onto an ITO surface through a nanoporous alumina mask. This new combined technique showed a broader distribution of hot spots and a higher signal-to-noise ratio than current SERS techniques due to the highly reproducible and uniform geometrical structures over a large area. This SERS-active surface was applied as cell culture system to study living cells in situ within their culture environment without any external preparation processes. We applied this newly developed method to cell-based research to differentiate cell lines, cells at different cell cycle stages, and live/dead cells. The enhanced Raman signals achieved from each cell, which represent the changes in biochemical compositions, enabled differentiation of each state and the conditions of the cells. This SERS technique employing a tightly controlled nanostructure array can potentially be applied to single cell analysis, early cancer diagnosis and cell physiology research.

Show MeSH
Related in: MedlinePlus