Institute of Applied Physics, TU Wien, Getreidemarkt 9, A-1060 Vienna, Austria.
Department of Microbiology, University of Innsbruck, Technikerstraße 25, A-6020 Innsbruck, Austria.
Molecules. 2020 Jul 13;25(14):3199. doi: 10.3390/molecules25143199.
Single-molecule localization microscopy has boosted our understanding of biological samples by offering access to subdiffraction resolution using fluorescence microscopy methods. While in standard mammalian cells this approach has found wide-spread use, its application to filamentous fungi has been scarce. This is mainly due to experimental challenges that lead to high amounts of background signal because of ample autofluorescence. Here, we report the optimization of labeling, imaging and data analysis protocols to yield the first single-molecule localization microscopy images of the filamentous fungus . As an example, we show the spatial distribution of the Sur7 tetraspanin-family protein Sfp2 required for hyphal growth and cell wall stability in this mycoparasitic fungus.
单分子定位显微镜通过荧光显微镜方法提供了亚衍射分辨率的方法，从而提高了我们对生物样本的理解。虽然这种方法在标准哺乳动物细胞中得到了广泛应用，但在丝状真菌中的应用却很少。这主要是由于实验挑战导致大量背景信号，因为丝状真菌有丰富的自发荧光。在这里，我们报告了标记、成像和数据分析方案的优化，以获得丝状真菌的第一个单分子定位显微镜图像。例如，我们展示了丝状真菌中菌丝生长和细胞壁稳定性所必需的 Sur7 四跨膜蛋白家族蛋白 Sfp2 的空间分布。
