Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA.
Biophys J. 2011 May 18;100(10):2522-9. doi: 10.1016/j.bpj.2011.03.045.
We demonstrate electron microscopy of fully hydrated eukaryotic cells with nanometer resolution. Living Schizosaccharomyces pombe cells were loaded in a microfluidic chamber and imaged in liquid with scanning transmission electron microscopy (STEM). The native intracellular (ultra)structures of wild-type cells and three different mutants were studied without prior labeling, fixation, or staining. The STEM images revealed various intracellular components that were identified on the basis of their shape, size, location, and mass density. The maximal achieved spatial resolution in this initial study was 32 ± 8 nm, an order of magnitude better than achievable with light microscopy on pristine cells. Light-microscopy images of the same samples were correlated with the corresponding electron-microscopy images. Achieving synergy between the capabilities of light and electron microscopy, we anticipate that liquid STEM will be broadly applied to explore the ultrastructure of live cells.
我们展示了具有纳米分辨率的全水合真核细胞的电子显微镜。将活酿酒酵母细胞加载到微流控室中，并在液体中用扫描透射电子显微镜（STEM）进行成像。对野生型细胞和三种不同突变体的天然细胞内（超）结构进行了研究，而无需进行预先标记、固定或染色。STEM 图像显示了各种细胞内成分，这些成分是根据它们的形状、大小、位置和质量密度来识别的。在这项初步研究中，实现的最大空间分辨率为 32 ± 8nm，比在原始细胞上用光学显微镜可达到的分辨率好一个数量级。对相同样品的明场显微镜图像与相应的电子显微镜图像进行了关联。通过实现光镜和电子显微镜能力的协同作用，我们预计液体 STEM 将被广泛应用于探索活细胞的超微结构。
