The visualization of cellular ultrastructure over a wide range of volumes is becoming possible by increasingly powerful techniques grouped under the rubric “volume electron microscopy” or volume EM (vEM). Focused ion beam scanning electron microscopy (FIB-SEM) occupies a “Goldilocks zone” in vEM: iterative and automated cycles of milling and imaging allow the interrogation of microns-thick specimens in 3-D at resolutions of tens of nanometers or less. This bestows on FIB-SEM the unique ability to aid the accurate and precise study of architectures of virus-cell interactions. Here we give the virologist or cell biologist a primer on FIB-SEM imaging in the context of vEM and discuss practical aspects of a room temperature FIB-SEM experiment. In an in vitro study of SARS-CoV-2 infection, we show that accurate quantitation of viral densities and surface curvatures enabled by FIB-SEM imaging reveals SARS-CoV-2 viruses preferentially located at areas of plasma membrane that have positive mean curvatures.
【저자키워드】 SARS-CoV-2, Segmentation, FIB-SEM, volumeEM, vEM, virological synapse, 【초록키워드】 viruses, SARS-COV-2 infection, SARS-CoV-2 virus, electron microscopy, Viral, Microscopy, Visualization, automated, temperature, experiment, ultrastructure, interactions, cellular, room temperature, scanning electron microscopy, beam, Volume, ion beam, specimen, SARS-CoV-2 viruses, in vitro study, primer, plasma membrane, quantitation, positive, Cell, less, unique, reveal, iterative, increasingly, 【제목키워드】 Other, architecture, electron, Virologist,