AFM-STED correlative nanoscopy reveals a dark side in fluorescence microscopy imaging
AFM-STED correlative nanoscopy reveals a dark side in fluorescence microscopy imaging
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DOI：
10.1126/sciadv.aav8062
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发表时间：
2019-06-01
期刊：
影响因子：
13.6
通讯作者：
Diaspro, Alberto
中科院分区：
文献类型：
--
作者：
Cosentino, Michela;Canale, Claudio;Diaspro, Alberto
已知荧光团的存在可以影响分子过程的动力学。尽管如此，一个负担得起的技术来控制样品内的荧光团分布，以及由荧光团本身引起的不可预测的异常过程的上升，是失踪。我们将受激发射损耗（STED）显微镜与原子力显微镜相结合来研究淀粉样蛋白聚集体的形成。特别地，我们研究了胰岛素和两种同种异型的B淀粉样肽的体外聚集。我们遵循标准方法来诱导聚集并以不同的染料与蛋白质比率标记分子。只有一小部分的纤维状聚集体显示在STED图像中，表明标记的分子没有不明显地参与聚集过程。这一发现表明，标记的分子仅遵循聚集反应中存在的多种聚集途径中的选定途径。
It is known that the presence of fluorophores can influence the dynamics of molecular processes. Despite this, an affordable technique to control the fluorophore distribution within the sample, as well as the rise of unpredictable anomalous processes induced by the fluorophore itself, is missing. We coupled a stimulated emission depletion (STED) microscope with an atomic force microscope to investigate the formation of amyloid aggregates. In particular, we studied the in vitro aggregation of insulin and two alloforms of b amyloid peptides. We followed standard methods to induce the aggregation and to label the molecules at different dye-to-protein ratios. Only a fraction of the fibrillar aggregates was displayed in STED images, indicating that the labeled molecules did not participate indistinctly to the aggregation process. This finding demonstrates that labeled molecules follow only selected pathways of aggregation, among the multiple that are present in the aggregation reaction.