Institute of Chemical Technologies and Analytics, TU Wien, Vienna 1060, Austria.
Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Vienna 1060, Austria.
Anal Chem. 2020 Dec 15;92(24):15719-15725. doi: 10.1021/acs.analchem.0c02228. Epub 2020 Dec 1.
Determination of the intracellular location of proteins is one of the fundamental tasks of microbiology. Conventionally, label-based microscopy and super-resolution techniques are employed. In this work, we demonstrate a new technique that can determine intracellular protein distribution at nanometer spatial resolution. This method combines nanoscale spatial resolution chemical imaging using the photothermal-induced resonance (PTIR) technique with multivariate modeling to reveal the intracellular distribution of cell components. Here, we demonstrate its viability by imaging the distribution of major cellulases and xylanases in using the colocation of a fluorescent label (enhanced yellow fluorescence protein, EYFP) with the target enzymes to calibrate the chemometric model. The obtained partial least squares model successfully shows the distribution of these proteins inside the cell and opens the door for further studies on protein secretion mechanisms using PTIR.
确定蛋白质的细胞内位置是微生物学的基本任务之一。传统上，采用基于标记的显微镜和超分辨率技术。在这项工作中，我们展示了一种可以以纳米空间分辨率确定细胞内蛋白质分布的新技术。该方法结合了使用光热诱导共振（PTIR）技术的纳米级空间分辨率化学成像以及多元建模，以揭示细胞成分的细胞内分布。在这里，我们通过用荧光标记（增强型黄色荧光蛋白，EYFP）与目标酶共定位来对细胞成分进行成像，从而证明了该方法的可行性，以校准化学计量模型。获得的偏最小二乘模型成功地显示了这些蛋白质在细胞内的分布，并为使用 PTIR 研究蛋白质分泌机制开辟了道路。
