Cellular imaging by targeted assembly of hot-spot SERS and photoacoustic nanoprobes using split-fluorescent protein scaffolds.
Cellular imaging by targeted assembly of hot-spot SERS and photoacoustic nanoprobes using split-fluorescent protein scaffolds.
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DOI：
10.1038/s41467-018-03046-w
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发表时间：
2018-02-09
影响因子：
16.6
通讯作者：
Pinaud F
中科院分区：
文献类型：
--
作者：
Köker T;Tang N;Tian C;Zhang W;Wang X;Martel R;Pinaud F
等离子体纳米材料在细胞内组装成光响应探针对于许多生物成像和纳米光子应用是非常感兴趣的，但对于传统的基于核酸支架的自下而上方法仍然具有挑战性。在这里，我们使用分裂荧光蛋白（FP）片段作为分子胶水和可切换的拉曼报告组装金或银等离子体纳米粒子（NP）直接在活细胞中的光子簇来解决这个难题。当靶向扩散癌细胞中的表面生物标志物时，NP自组装成表面增强拉曼散射（Sers）纳米簇，其具有通过全长FP的重建均匀接种的热点。在等离子体热点内，FP发色团的自催化活化和其拉曼指纹的近场放大使得能够对靶细胞进行选择性和灵敏的Sers成像。这种FP驱动的金属胶体组装也产生增强的光声信号，允许混合FP/NP纳米团簇用作具有单细胞灵敏度的多峰Sers和光声显微镜的造影剂。用于将等离子体纳米颗粒组装到光响应探针中的传统方法存在多个问题。在这里，作者使用分裂的荧光蛋白片段作为分子胶，形成稳定的纳米团簇，用于活细胞中的表面增强拉曼散射和光声成像。
The in cellulo assembly of plasmonic nanomaterials into photo-responsive probes is of great interest for many bioimaging and nanophotonic applications but remains challenging with traditional nucleic acid scaffolds-based bottom-up methods. Here, we address this quandary using split-fluorescent protein (FP) fragments as molecular glue and switchable Raman reporters to assemble gold or silver plasmonic nanoparticles (NPs) into photonic clusters directly in live cells. When targeted to diffusing surface biomarkers in cancer cells, the NPs self-assemble into surface-enhanced Raman-scattering (SERS) nanoclusters having hot spots homogenously seeded by the reconstruction of full-length FPs. Within plasmonic hot spots, autocatalytic activation of the FP chromophore and near-field amplification of its Raman fingerprints enable selective and sensitive SERS imaging of targeted cells. This FP-driven assembly of metal colloids also yields enhanced photoacoustic signals, allowing the hybrid FP/NP nanoclusters to serve as contrast agents for multimodal SERS and photoacoustic microscopy with single-cell sensitivity. Traditional methods for the assembly of plasmonic nanoparticles into photo-responsive probes suffer from multiple problems. Here the authors use split fluorescent protein fragments as molecular glue to form stable nanoclusters for surface enhanced Raman scattering and photoacoustic imaging in live cells.
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