High-Resolution Correlative Microscopy: Bridging the Gap between Single Molecule Localization Microscopy and Atomic Force Microscopy
High-Resolution Correlative Microscopy: Bridging the Gap between Single Molecule Localization Microscopy and Atomic Force Microscopy
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DOI：
10.1021/acs.nanolett.5b00572
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发表时间：
2015-08-01
期刊：
影响因子：
10.8
通讯作者：
Fantner, Georg E.
中科院分区：
文献类型：
--
作者：
Odermatt, Pascal D.;Shivanandan, Arun;Fantner, Georg E.
活体样品的纳米级表征对于现代生物学来说已经变得至关重要。原子力显微镜（AFM）创建从生物分子到水环境中活细胞的脆弱生物结构的拓扑图像。然而，将纳米级结构与特定蛋白质的生物学功能相关联可能具有挑战性。为此，我们已经建立了一个相关的单分子定位显微镜（SMLM）/AFM，允许本地化特定的，标记的蛋白质在高分辨率的AFM图像在生物相关的上下文中。使用直接随机光学重建显微镜（dSTORM）/原子力显微镜，我们直接相关和量化的密度与三维地形使用两种成像方式沿着（F-）肌动蛋白细胞骨架丝的本地化。此外，使用光激活光显微镜（PALM）/AFM，我们提供了相关的图像，细菌细胞在水性条件下。此外，我们报告的第一个相关的AFM/PALM成像活的哺乳动物细胞。这两种技术提供的互补信息为结构和功能纳米生物学开辟了新的维度。
Nanoscale characterization of living samples has become essential for modern biology. Atomic force microscopy (AFM) creates topological images of fragile biological structures from biomolecules to living cells in aqueous environments. However, correlating nanoscale structure to biological function of specific proteins can be challenging. To this end we have built and characterized a correlated single molecule localization microscope (SMLM)/AFM that allows localizing specific, labeled proteins within high-resolution AFM images in a biologically relevant context. Using direct stochastic optical reconstruction microscopy (dSTORM)/AFM, we directly correlate and quantify the density of localizations with the 3D topography using both imaging modalities along (F-)actin cytoskeletal filaments. In addition, using photo activated light microscopy (PALM)/AFM, we provide correlative images of bacterial cells in aqueous conditions. Moreover, we report the first correlated AFM/PALM imaging of live mammalian cells. The complementary information provided by the two techniques opens a new dimension for structural and functional nanoscale biology.