Abstract
Manual selection of targets in experimental or diagnostic samples by transmission electron microscopy (TEM), based on single overview and detail micrographs, has been time-consuming and susceptible to bias. Substantial information and throughput gain may now be achieved by the automated acquisition of virtually all structures in a given EM section. Resulting datasets allow the convenient pan-and-zoom examination of tissue ultrastructure with preserved microanatomical orientation. The technique is, however, critically sensitive to artifacts in sample preparation. We, therefore, established a methodology to prepare large-scale digitization samples (LDS) designed to acquire entire sections free of obscuring flaws. For evaluation, we highlight the supreme performance of scanning EM in transmission mode compared with other EM technology. The use of LDS will substantially facilitate access to EM data for a broad range of applications.
Highlights
Electron microscopy (EM) continues to be a valuable tool in basic research and anatomical pathology
The traditional workflow for transmission EM (TEM) with an operator interactively screening specimens at low magnification and selecting targets of interest for snapshots at high resolution may, be timeconsuming, laborious, and sensitive to bias so that its use became diminished over the past decades despite its undisputed values (Lee & Mak, 2011; de Haro & Furness, 2012)
The reliable and easy-to-implement methodology for the preparation of ultrathin sections was to be adapted for large-scale digitization with modern scanning EM (SEM)-STEM or transmission electron microscopy (TEM) systems, serving to analyze a broad range of native or experimental tissues and cells
Summary
Electron microscopy (EM) continues to be a valuable tool in basic research and anatomical pathology. Aside from educated inspection of regions of interest (ROI) by the operator, EM traditionally serves to detect unexpected features in cells and tissues in an “open view” manner (Pavlisko & Howell, 2013). EM allows flexible recording of structures that may vary in size by three orders of magnitude, delivering excellent resolution down to the nanometer scale. The traditional workflow for transmission EM (TEM) with an operator interactively screening specimens at low magnification and selecting targets of interest for snapshots at high resolution may, be timeconsuming, laborious, and sensitive to bias so that its use became diminished over the past decades despite its undisputed values (Lee & Mak, 2011; de Haro & Furness, 2012). Consequent loss of expertise may lead to misinterpretations as recently reported for coronavirus particle structure in SARS-CoV-2 pandemic (de Haro & Furness, 2012; Dittmayer et al, 2020; Bullock et al, 2021)
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