Methodology for sample preparation and size measurement of commercial ZnO nanoparticles.

This study discusses the strategies on sample preparation to acquire images with sufficient quality for size characterization by scanning electron microscope (SEM) using two commercial ZnO nanoparticles of different surface properties as a demonstration. The central idea is that micrometer sized aggregates of ZnO in powdered forms need to firstly be broken down to nanosized particles through an appropriate process to generate nanoparticle dispersion before being deposited on a flat surface for SEM observation. Analytical tools such as contact angle, dynamic light scattering and zeta potential have been utilized to optimize the procedure for sample preparation and to check the quality of the results. Meanwhile, measurements of zeta potential values on flat surfaces also provide critical information and save lots of time and efforts in selection of suitable substrate for particles of different properties to be attracted and kept on the surface without further aggregation. This simple, low-cost methodology can be generally applied on size characterization of commercial ZnO nanoparticles with limited information from vendors.

A new method using Scanning Electron Microscopy (SEM) for preparation of anisopterous odonates.

Anisopterous odonate male's secondary genitalia is a complex of several structures, among them the vesica spermalis is the most informative with important specific characters. The observation of those characters, mostly of membranous nature, is difficult in the Scanning Electron Microscope due to dehydration and metallization processes. In this contribution, we discuss a new and low cost procedure for the observation of these characters in the SEM, compatible with the most common agents used for preserving specimens.

3D printing scanning electron microscopy sample holders: A quick and cost effective alternative for custom holder fabrication.

A simple and cost effective alternative for fabricating custom Scanning Electron Microscope (SEM) sample holders using 3D printers and conductive polylactic acid filament is presented. The flexibility of the 3D printing process allowed for the fabrication of sample holders with specific features that enable the high-resolution imaging of nanoelectrodes and nanopipettes. The precise value of the inner semi cone angle of the nanopipettes taper was extracted from the acquired images and used for calculating their radius using electrochemical methods. Because of the low electrical resistivity presented by the 3D printed holder, the imaging of non-conductive nanomaterials, such as alumina powder, was found to be possible. The fabrication time for each sample holder was under 30 minutes and the average cost was less than $0.50 per piece. Despite being quick and economical to fabricate, the sample holders were found to be sufficiently resistant, allowing for multiple uses of the same holder.

A high performance, cost-effective, open-source microscope for scanning two-photon microscopy that is modular and readily adaptable.

Two-photon laser scanning microscopy has revolutionized the ability to delineate cellular and physiological function in acutely isolated tissue and in vivo. However, there exist barriers for many laboratories to acquire two-photon microscopes. Additionally, if owned, typical systems are difficult to modify to rapidly evolving methodologies. A potential solution to these problems is to enable scientists to build their own high-performance and adaptable system by overcoming a resource insufficiency. Here we present a detailed hardware resource and protocol for building an upright, highly modular and adaptable two-photon laser scanning fluorescence microscope that can be used for in vitro or in vivo applications. The microscope is comprised of high-end componentry on a skeleton of off-the-shelf compatible opto-mechanical parts. The dedicated design enabled imaging depths close to 1 mm into mouse brain tissue and a signal-to-noise ratio that exceeded all commercial two-photon systems tested. In addition to a detailed parts list, instructions for assembly, testing and troubleshooting, our plan includes complete three dimensional computer models that greatly reduce the knowledge base required for the non-expert user. This open-source resource lowers barriers in order to equip more laboratories with high-performance two-photon imaging and to help progress our understanding of the cellular and physiological function of living systems.

Automated transmission-mode scanning electron microscopy (tSEM) for large volume analysis at nanoscale resolution.

Transmission-mode scanning electron microscopy (tSEM) on a field emission SEM platform was developed for efficient and cost-effective imaging of circuit-scale volumes from brain at nanoscale resolution. Image area was maximized while optimizing the resolution and dynamic range necessary for discriminating key subcellular structures, such as small axonal, dendritic and glial processes, synapses, smooth endoplasmic reticulum, vesicles, microtubules, polyribosomes, and endosomes which are critical for neuronal function. Individual image fields from the tSEM system were up to 4,295 µm(2) (65.54 µm per side) at 2 nm pixel size, contrasting with image fields from a modern transmission electron microscope (TEM) system, which were only 66.59 µm(2) (8.160 µm per side) at the same pixel size. The tSEM produced outstanding images and had reduced distortion and drift relative to TEM. Automated stage and scan control in tSEM easily provided unattended serial section imaging and montaging. Lens and scan properties on both TEM and SEM platforms revealed no significant nonlinear distortions within a central field of ∼100 µm(2) and produced near-perfect image registration across serial sections using the computational elastic alignment tool in Fiji/TrakEM2 software, and reliable geometric measurements from RECONSTRUCT™ or Fiji/TrakEM2 software. Axial resolution limits the analysis of small structures contained within a section (∼45 nm). Since this new tSEM is non-destructive, objects within a section can be explored at finer axial resolution in TEM tomography with current methods. Future development of tSEM tomography promises thinner axial resolution producing nearly isotropic voxels and should provide within-section analyses of structures without changing platforms. Brain was the test system given our interest in synaptic connectivity and plasticity; however, the new tSEM system is readily applicable to other biological systems.

SEM in service pathology: a review of its potential role.

New medical technology evolves through distinct phases. Initial technical proving is followed by a phase of detailed data recording, often with no detailed attempt to fill any specific service requirements. Next an attempt is made to establish correlations with other techniques and results, with a view to defining any unique attributes of the technology in particular fields. Detailed service-related applications are then worked out, but the final adoption of new instrumentation into clinical practice depends not only on the success of its technical performance, but also on its cost-effectiveness. This paper reviews studies of clinically-derived material from the viewpoint of a U.K. diagnostic pathologist. Detailed comparisons are drawn between the evolution of SEM on the one hand and of LM and CTEM on the other. An attempt is made to identify areas of practice in which SEM might make the greatest impact in the foreseeable future. It is proposed that paraffin-embedded tissue in pathology back files may provide a valuable source of investigative material for diagnostic SEM.