Raman spectroscopy is becoming increasingly popular as an in vitro bioimaging technique, when coupled with plasmonic substrates such as gold nanoparticles (AuNPs). Plasmonic AuNPs not only display excellent biocompatibility but can also induce the surface‐enhanced Raman scattering (SERS) effect, which can be exploited for cell labeling, as an interesting alternative to fluorescence‐based techniques. SERS bioimaging requires the use of so‐called SERS tags or SERS‐encoded AuNPs. A remaining difficulty toward the general implementation of this method is the difficulty to correlate the SERS signal (spectral intensity) with the number of SERS tags. Therefore, a general correlation method, suitable for arbitrary AuNP morphologies and Raman‐active molecules (Raman reporters or RaRs), should largely improve the quantitative character of SERS as an imaging technique. We propose a protocol, with an associated app (SERSTEM), which enables the user to determine the average SERS intensity per nanoparticle from transmission electron microscopy (TEM) and SERS data. As a proof of concept, we demonstrated the method for Au nanostars and nanorods, carrying four different RaRs, and implemented the SERSTEM app, which is publicly available from an open‐source platform.

中文翻译：

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SERSTEM：用于对相关SERS和TEM成像进行统计分析并评估SERS标签性能的应用程序

拉曼光谱法与诸如金纳米颗粒（AuNPs）的等离激元底物结合后，作为一种体外生物成像技术正变得越来越流行。等离子体金纳米颗粒不仅显示出出色的生物相容性，而且还可以诱导表面增强拉曼散射（SERS）效应，可以将其用于细胞标记，作为基于荧光技术的有趣替代方法。SERS生物成像需要使用所谓的SERS标签或SERS编码的AuNP。对该方法的一般实现的另一个困难是难以将SERS信号（频谱强度）与SERS标签的数量相关联。因此，一种适用于任意AuNP形态和拉曼活性分子（拉曼报告分子或RaRs）的通用相关方法，应该大大改善SERS作为成像技术的定量特征。我们提出了一个带有相关应用程序（SERSTEM）的协议，该协议使用户能够从透射电子显微镜（TEM）和SERS数据确定每个纳米颗粒的平均SERS强度。作为概念验证，我们演示了带有四个不同RaR的Au纳米星和纳米棒的方法，并实现了SERSTEM应用程序，该应用程序可从开源平台上公开获得。