Plasmonic Nanoprobes for in Vivo Multimodal Sensing and Bioimaging of MicroRNA within Plants
Plasmonic Nanoprobes for in Vivo Multimodal Sensing and Bioimaging of MicroRNA within Plants
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DOI：
10.1021/acsami.8b19977
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发表时间：
2019-02-27
影响因子：
9.5
通讯作者：
Vo-Dinh, Tuan
中科院分区：
文献类型：
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作者：
Crawford, Bridget M.;Strobbia, Pietro;Vo-Dinh, Tuan
从植物生物学到农业生物技术和生物燃料开发，监测整个植物中的基因表达是至关重要的；然而，目前还没有一种方法可以在不需要提取样本的情况下对植物系统中的基因组靶标进行活体监测。在这里，我们报道了一种基于等离子体纳米探针的独特的多模式方法，通过集成三种不同和互补的技术：表面增强拉曼散射(SERS)、X射线荧光(XRF)和等离子体增强双光子发光(TPL)，能够对整个植物叶片中的microRNA生物目标进行活体成像和生物传感。由于金的含量和较高的双光子发光截面，开发的方法使用等离子体纳米星，不仅提供了很大的拉曼信号增强，而且允许通过XRF和等离子体增强的TPL进行定位和定量。我们的方法使用反向分子哨兵纳米探针对拟南芥叶片中的microRNA进行SERS生物成像，以提供检测到的microRNA目标的动态SERS图，同时还使用XRF和TPL来定量纳米探针的浓度。观察到纳米探针在渗透到叶片组织时占据了细胞间的空间。这份报告为使用等离子体纳米探针对整个植物的核酸生物靶标进行体内功能成像奠定了基础，这一工具将以前所未有的空间和时间分辨率研究这些生物靶标，分别达到200微米和30分钟，从而为生物工程研究带来革命性的变化。
Monitoring gene expression within whole plants is critical for many applications ranging from plant biology to agricultural biotechnology and biofuel development; however, no method currently exists for in vivo monitoring of genomic targets in plant systems without requiring sample extraction. Herein, we report a unique multimodal method based on plasmonic nanoprobes capable of in vivo imaging and biosensing of microRNA biotargets within whole plant leaves by integrating three different and complementary techniques: surface-enhanced Raman scattering (SERS), X-ray fluorescence (XRF), and plasmonics-enhanced two-photon luminescence (TPL). The method developed uses plasmonic nanostars, which not only provide large Raman signal enhancement but also allow for localization and quantification by XRF and plasmonics-enhanced TPL, owing to gold content and high two-photon luminescence cross sections. Our method uses inverse molecular sentinel nanoprobes for SERS bioimaging of microRNA within Arabidopsis thaliana leaves to provide a dynamic SERS map of detected microRNA targets while also quantifying nanoprobe concentrations using XRF and TPL. The nanoprobes were observed to occupy the intercellular spaces upon infiltration into the leaf tissues. This report lays the foundation for the use of plasmonic nanoprobes for in vivo functional imaging of nucleic acid biotargets in whole plants, a tool that will revolutionize bioengineering research by allowing the study of these biotargets with previously unmet spatial and temporal resolution, 200 mu m and 30 min, respectively.