Scanning Transmission Electron Microscopy in a Scanning Electron Microscope for the High-Throughput Imaging of Biological Assemblies,Biomacromolecules

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Scanning Transmission Electron Microscopy in a Scanning Electron Microscope for the High-Throughput Imaging of Biological Assemblies
Biomacromolecules ( IF 5.5 ) Pub Date : 2022-07-26 , DOI: 10.1021/acs.biomac.2c00323
Kelly A Parker ₁ , Stephanie Ribet ₁ , Blaise R Kimmel ₂ , Roberto Dos Reis _{1,

3} , Milan Mrksich _{4,

5} , Vinayak P Dravid _{1,

3}

Affiliation

Electron microscopy of soft and biological materials, or “soft electron microscopy”, is essential to the characterization of macromolecules. Soft microscopy is governed by enhancing contrast while maintaining low electron doses, and sample preparation and imaging methodologies are driven by the length scale of features of interest. While cryo-electron microscopy offers the highest resolution, larger structures can be characterized efficiently and with high contrast using low-voltage electron microscopy by performing scanning transmission electron microscopy in a scanning electron microscope (STEM-in-SEM). Here, STEM-in-SEM is demonstrated for a four-lobed protein assembly where the arrangement of the proteins in the construct must be examined. STEM image simulations show the theoretical contrast enhancement at SEM-level voltages for unstained structures, and experimental images with multiple STEM modes exhibit the resolution possible for negative-stained proteins. This technique can be extended to complex protein assemblies, larger structures such as cell sections, and hybrid materials, making STEM-in-SEM a valuable high-throughput imaging method.

中文翻译：

扫描电子显微镜中的扫描透射电子显微镜用于生物组件的高通量成像

软和生物材料的电子显微镜或“软电子显微镜”对于表征大分子至关重要。软显微镜通过在保持低电子剂量的同时增强对比度来控制，样品制备和成像方法由感兴趣特征的长度尺度驱动。虽然冷冻电子显微镜提供最高分辨率，但通过在扫描电子显微镜 (STEM-in-SEM) 中执行扫描透射电子显微镜，可以使用低压电子显微镜以高对比度有效地表征较大的结构。在这里，STEM-in-SEM 被证明用于四叶蛋白质组装，其中必须检查构建体中蛋白质的排列。STEM 图像模拟显示了未染色结构在 SEM 级电压下的理论对比度增强，并且具有多种 STEM 模式的实验图像显示了负染色蛋白质可能的分辨率。该技术可以扩展到复杂的蛋白质组装、更大的结构（如细胞切片）和混合材料，使 STEM-in-SEM 成为一种有价值的高通量成像方法。

更新日期：2022-07-26

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