Development and application of STEM for the biological sciences

作者：Sousa Alioscka A; Leapman Richard D^*

来源：Ultramicroscopy, 2012, 123: 38-49.

DOI：10.1016/j.ultramic.2012.04.005

• Scanning transmission electron microscopy (STEM)
• Mass mapping
• Macromolecular assemblies
• Graphene
• Gold nanoparticles
• Electron energy loss spectroscopy (EELS)
• Electron tomography
• Cellular structure

The design of the scanning transmission electron microscope (STEM), as conceived originally by Crewe and coworkers, enables the highly efficient and flexible collection of different elastic and inelastic signals resulting from the interaction of a focused probe of incident electrons with a specimen. In the present paper we provide a brief review for how the STEM today can be applied towards a range of different problems in the biological sciences, emphasizing four main areas of application. (1) For three decades, the most widely used STEM technique has been the mass determination of proteins and other macromolecular assemblies. Such measurements can be performed at low electron dose by collecting the high-angle dark-field signal using an annular detector. STEM mass mapping has proven valuable for characterizing large protein assemblies such as filamentous proteins with a well-defined mass per length.(2) The annular dark-field signal can also be used to image ultra small, functionalized nanoparticles of heavy atoms for labeling specific amino-acid sequences in protein assemblies. (3) By acquiring electron energy loss spectra (EELS) at each pixel in a hyperspectral image, it is possible to map the distributions of specific bound elements like phosphorus, calcium and iron in isolated macromolecular assemblies or in compartments within sectioned cells. Near single atom sensitivity is feasible provided that the specimen can tolerate a very high incident electron dose.( 4) Electron tomography is a new application of STEM that enables three-dimensional reconstruction of micrometer-thick sections of cells. In this technique a probe of small convergence angle gives a large depth of field through out the thickness of the specimen while maintaining a probe diameter of <2 nm; and the use of an on-axis bright-field detect or reduces the effects of beam broadening and thus improves the spatial resolution compared to that attainable by STEM dark-field tomography. Published by Elsevier B.V.

• 出版日期2012-12