Imaging the uptake of gold nanoshells in live cells using plasmon resonance enhanced four wave mixing microscopy.
Imaging the uptake of gold nanoshells in live cells using plasmon resonance enhanced four wave mixing microscopy.
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DOI：
10.1364/oe.19.017563
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发表时间：
2011-08-29
期刊：
影响因子：
3.8
通讯作者：
Moger J
中科院分区：
文献类型：
--
作者：
Garrett N;Whiteman M;Moger J
金纳米壳（GNS）是一种新型金属纳米颗粒，具有引人注目的光学特性，使其非常适合生物光子学应用。我们提出了一项新颖的研究，使用等离子体增强四波混合显微镜与相干反斯托克斯拉曼散射（CARS）显微镜相结合，以可视化活细胞内 75 nm 半径 GNS 的分布。在激光耐受性研究中，我们发现含有纳米壳的细胞可以暴露在 < 2.5 mJ 的能量下，没有检测到光热诱导的坏死，而细胞死亡与超过该阈值的功率成线性比例。检测到的大部分 GNS 信号来自等离子体增强四波混合 (FWM)，我们在外延方向上检测到，入射激光调谐到拉曼光谱的静默区域。通过将 Epi 检测到的信号与 CH2 共振处的前向检测到的 CARS 相结合，可以可视化细胞 GNS 分布。使用两种供体分子 NaHS 和 GYY4137 研究 H2S 对纳米壳吸收的影响，证明了该技术对现实世界纳米颗粒剂量问题的适用性。随着 GYY4137 浓度从 10 μM 增加到 1 mM，纳米壳像素百分比作为细胞体积的函数 (PPCV) 从 2.15% 增加到 3.77%。当NaHS浓度在相同范围内增加时，纳米壳PPCV从12.67%下降到11.47%。本研究发现影响吸收的最重要因素是 H2S 释放速率，NaHS 的快速释放导致吸收量显着增加。
Gold nanoshells (GNS) are novel metal nanoparticles exhibiting attractive optical properties which make them highly suitable for biophotonics applications. We present a novel investigation using plasmon-enhanced four wave mixing microscopy combined with coherent anti-Stokes Raman scattering (CARS) microscopy to visualize the distribution of 75 nm radius GNS within live cells. During a laser tolerance study we found that cells containing nanoshells could be exposed to < 2.5 mJ each with no photo-thermally induced necrosis detected, while cell death was linearly proportional to the power over this threshold. The majority of the GNS signal detected was from plasmon-enhanced four wave mixing (FWM) that we detected in the epi-direction with the incident lasers tuned to the silent region of the Raman spectrum. The cellular GNS distribution was visualized by combining the epi-detected signal with forwards-detected CARS at the CH2 resonance. The applicability of this technique to real-world nanoparticle dosing problems was demonstrated in a study of the effect of H2S on nanoshell uptake using two donor molecules, NaHS and GYY4137. As GYY4137 concentration was increased from 10 μM to 1 mM, the nanoshell pixel percentage as a function of cell volume (PPCV) increased from 2.15% to 3.77%. As NaHS concentration was increased over the same range, the nanoshell PPCV decreased from 12.67% to 11.47%. The most important factor affecting uptake in this study was found to be the rate of H2S release, with rapid-release from NaHS resulting in significantly greater uptake.