Different aspects of electron-beam damage are summarized, together with some quantitative evaluation. TEM and STEM are compared in terms of information-to-damage ratio. Electron-beam fabrication is briefly considered in terms of resolution and writing speed.

Recent years have seen increased crossover of microscopy techniques developed for transmission electron microscopes (TEMs) being utilized as well in scanning electron microscopes (SEMs). The order of magnitude lower beam energies available in SEMs correspond with significantly different beam-sample interactions that must be accounted for when examining freestanding films. This is especially important for composition analysis via energy dispersive X-ray spectroscopy (EDS) since lower-energy transmission experiments mean X-ray volumes become comparable to sample dimensions. To better understand this scenario, we report results of an SEM-EDS study of terraced freestanding films comprising binary compounds with well-defined atomic ratios. Guided by Monte Carlo simulations, EDS line scan data were collected at different beam energies across segments of varying thicknesses. The results of quantification analysis using different models are compared for multiple combinations of beam energy, sample thickness, and material density, from which some general guidelines are provided for SEM-EDS analysis of freestanding films.

In response to the requirements imposed by the COVID-19 pandemic in 2020, we developed a remote learning undergraduate workshop for 44 students at the University of Newcastle by embedding scanning electron microscope (SEM) images of Maratus (Peacock) spiders into the MyScope Explore environment. The workshop session had two main components: 1) to use the online MyScope Explore tool to virtually image scales with structural color and pigmented color on Maratus spiders; 2) to join a live SEM session via Zoom to image an actual Maratus spider. In previous years, the undergraduate university students attending this annual workshop would enter the Microscopy Facility at the University of Newcastle to image specimens with SEM; however, in 2020 the Microscopy Facility was closed to student visitors, and this virtual activity was developed in order to proceed with the educational event. The program was highly successful and constitutes a platform that can be used in the future by universities for teaching microscopy remotely.

The education of children, particularly in scientific inquiry, is a powerful force that has shaped humanity's history and holds the key to our future. Given their power to provoke wonder and awe in child and adult alike, microscopes have proven to be transformative as tools for education. Exploratory learning using microscopes has helped to overcome critical barriers in science, technology, engineering, and math (STEM) education and to broaden participation in these vital fields. Since its inception, the Microscopy Society of America (MSA) has recognized this and made education and outreach to children a priority. Here, we provide a brief review of MSA's efforts in this direction, focusing on current efforts aimed at improving STEM education through the greater adoption and utilization of microscopes.

Stimulated emission depletion (STED) microscopy is a powerful tool for the study of sub-micron samples, especially in biology. In this article, we show a simple, straightforward method for modeling a STED microscope using Zemax geometrical optics ray tracing principles, while still achieving a realistic spot size. This method is based on scattering models, and while it employs fast ray-tracing simulations, its output behavior is consistent with the real, diffractive behavior of a STED de-excitation spot. The method can be used with Zemax Black Boxes of real objectives for analysis of their performance in a STED setup and open perspective of modeling, tolerancing performance, and then building advanced microscope systems from scratch. In an example setup, we show an analysis method for both XY and Z excitation and de-excitation intensity distributions. This method can simplify the design of custom STED setups by proper modeling of the resolution performance.

The CytoViva Enhanced Darkfield (EDF) Illuminator enables direct optical observation of nano-scale entities in a wide range of transparent and translucent sample environments, without requiring the use of labels or other markers. The improved performance is a result of a uniquely designed light path, which focuses maximum photon density on the sample, thus enabling superior signal-to-noise imaging performance. This high-performance capability not only provides users with previously unobtainable images, but it also allows established techniques such as hyperspectral and Raman microscopy to be used in new and exciting ways.