Remarkable discoveries such as condensed chromatin in duckbill dinosaur cartilage and newly presented discoveries of dinosaur vascular veins, venule valves, and nerve fibers (this report) warrant study of global dinosaur remains for preserved soft tissue (dST). Dinosaur osteocytes feature dendritic projections (filipodia) of lengths up to 18 µm, while osteocytes collected from a Triceratops horn have lengths of 25 µm or longer. While preservation methods for dST involve the degradation of sugars into glycation end products and the employment of highly oxidative hydroxyl radicals to “fix” tissues, we note that lengthy and narrow osteocyte filipodia show no signs of hydroxyl radical infiltration into the lacuna-canalicular network. Moreover, our ultraviolet fluorescence (UVFL) study of Triceratops horn, rib, vertebra, and frill thin sections shows extensive clotting in most vessel canals, probably as a result of asphyxia while drowning. Further UVFL autofluorescence study of dinosaur bone sections is vital for characterization of dinosaur blood clots.

Based on technologies capable of data collection between the millimeter and nanometer scales, correlative imaging has been transforming how researchers obtain molecular and spatial information from specimens. Attempts to combine multidimensional data are often met with the challenge of overcoming suboptimal sample conditions such as reduced fluorescence signal, poor specimen preservation, anisotropic specimen deformation, and low specimen contrast. These issues motivated the development and use of enhanced sample preparation procedures, as well as specialized imaging software to overcome such challenges. In this work we present three simple methods to correlate optical and scanning electron microscopy images.

Technological advancements in the areas of sample illumination, image acquisition, and image processing have significantly improved the speed and sensitivity of fluorescence microscopy. In particular, light emitting diodes (LEDs) coupled to transistor-transistor logic (TTL) circuits have reduced photo-bleaching and photo-toxicity by limiting sample illumination to the camera exposure time. Unfortunately, many microscopes still rely on bulb-based light sources that cannot be configured with TTL. Moreover, even when TTL is enabled in conventional microscope software, hardware and software delays can still contribute to photo-toxicity and lead to additional delays between subsequent images, introducing errors in time lapse image recordings. The goal of the present article is to highlight the significance of these issues.

Lithium ion batteries (LIBs) are key components of modern electronics and are regularly utilized as their primary power source. Understanding the electrical and mechanical properties of electrode materials plays a major role in the performance improvement of LIBs. In this article, we provide research using PinPoint™ scanning spreading resistance microscopy (SSRM) to effectively measure both electrical and mechanical properties of LIB electrode surfaces at a much higher quality in a high-vacuum environment than in ambient conditions. The data collected in this experiment demonstrate that this technique is an effective means for measuring the quantitative and qualitative topographical, electrical and mechanical data of advanced materials with improved image quality and data accuracy.

Today, researchers have a number of powerful image processing methods at hand that have the potential to make comparably simple and inexpensive widefield microscope systems more powerful and versatile. These techniques can enable new insights into biological samples and allow new discoveries, so long as their limitations and pitfalls are known.

Monte Carlo simulations are commonly used in elemental quantification using energy-dispersive spectroscopy (EDS). Here, the Monte Carlo program MC X-ray was incorporated into the image processing software Dragonfly by Object Research Systems (ORS) as a simulation library. The simulation program has been transformed into a complete microscope simulator where the tools of Dragonfly allow complex voxel-based geometries to be constructed, and the electron beam and detectors can be freely placed inside the 3D space. Computation times of simulations have been improved drastically through new data structures and parallelization. Simulations of backscattered electron imaging and EDS mapping are presented here to demonstrate the capabilities of this new library.

A method was developed to expedite the imaging of diatoms by optimizing their orientation. This quick technique puts grooves into a SEM stub so that as the diatoms are laid down they either land on the flat surface or in a variety of positions in the grooves, thereby reducing time required to capture taxonomically critical features.

Electron diffraction is an essential tool for materials scientists to have in their characterization toolboxes. When using the transmission electron microscope (TEM) to perform diffraction experiments, setting up the microscope for both calibration standards and unknown materials in a consistent method will ensure that dependable results are obtained. Care must also be exercised to protect digital cameras from intense transmitted and diffracted beams to avoid damage. A procedure is presented that will allow a microscopist to reproducibly configure the microscope and sample to acquire selected area electron diffraction patterns so that these issues are addressed.

The Online Electron Microscopy Platform makes electron microscopy education accessible by combining simulation-based practice with a range of educational content. Students learn how to operate electron microscopes by using virtual microscopes that precisely simulate the functions of real microscopes. Built as a web application, the platform can be used by an unlimited number of people and at any time. This platform reduces the time needed for training users to operate a real microscope. By preparing students in STEM disciplines to use electron microscopes, the system helps them acquire the skills they need to succeed in the modern workforce.

In the 50 years since the first mating of semiconductor-based energy-dispersive X-ray spectrometry (EDS) with the scanning electron microscope (SEM), this hybrid instrument has become an indispensable microanalytical tool. In the last two decades a new detector, the silicon drift detector (SDD), has overtaken earlier Si(Li) technology and has made EDS in the SEM and TEM faster and better. This article tells the story of the SDD development and describes improvements in count rate capability, energy resolution, and detector geometry that bring to SEM microanalysis exceptional precision and stability. Quality maps of element distributions can now be obtained in minutes instead of hours.

Electron diffraction is an essential tool for materials scientists to have in their characterization toolboxes. When using the transmission electron microscope (TEM) to perform diffraction experiments, setting up the microscope for both calibration standards and unknown materials in a consistent method will ensure that dependable results are obtained. Care must also be exercised to protect digital cameras from intense transmitted and diffracted beams to avoid damage. In Part 2 of this series a procedure is presented for the proper recording of SAED patterns.

Dynamic imaging—acquiring images and simultaneously recording relevant sensor parameters—is a powerful way to extend the information and insight available over and above optical microscopy. It improves understanding and correlates how properties of materials alter with, for example, changing temperature, pressure, humidity, tensile forces, shear stress, or aging. The basics of an imaging setup are the microscope, a relevant stage, a camera of appropriate performance, and suitable software to acquire, synchronize, and analyze the images and data collected. However, many factors need to be considered and optimized to ensure reliable experiments. These include basic features of the microscope—objective resolution and magnification, for example, as well as the accuracy and reproducibility of the stage, the camera format, dynamic range, and resolution. In this tutorial, we discuss the components of a dynamic data and image capture system and present examples where researchers have used this approach to better understand their materials.

In microscopy and analytical instrumentation, a researcher or clinician may need to excite several fluorophores in a sample in order to generate a useful fluorescence map of the cell or tissue of interest. This typically would include a nuclear marker such as DAPI, a green-emitting fluorophore such as FITC or GFP, and a red-emitting fluorophore such as mCherry, TRITC, or Texas Red. These red-emitting dyes can be efficiently excited using an arc lamp with a strong peak in the 550 nm region and another at 580 nm. When technology in microscope light sources originally moved to light-emitting diodes (LEDs) in the early 2000s, fluorescence work in the green gap (540 to 590 nm) excitation range was a challenge with no solution. This article explains how technology has adapted to provide LEDs that match the excitation of fluorophores typically used in multiplex fluorescence imaging.

While conventional negative stain electron microscopy (NSEM) has been used extensively for imaging and characterizing high density lipoprotein (HDL) particles, traditional methods have not provided high-level particle surface detail. To obtain greater detail of HDL particles, we developed a modified negative staining procedure using a uranyl formate staining solution that maintains particle morphology and provides improved surface detail. The additional surface detail has allowed us to perform single-particle analysis on whole HDL particle preparations. It has also allowed us to explore interactions between HDL particle surface proteins such as Apo A with proteins that interact with Apo A such as lecithin-cholesterol acyltransferase (LCAT).

Biological imaging tools continue to increase in speed, scale, and resolution, often resulting in the collection of gigabytes or even terabytes of data in a single experiment. In comparison, the ability of research laboratories to store and manage this data is lagging greatly. This leads to limits on the collection of valuable data and slows data analysis and research progress. Here we review common ways researchers store data and outline the drawbacks and benefits of each method. We also offer a blueprint and budget estimation for a currently deployed data server used to store large datasets from zebrafish brain activity experiments using light-sheet microscopy. Data storage strategy should be carefully considered and different options compared when designing imaging experiments.