In recent years the development of remote microscopy, specifically in electron microscopes, has begun to emerge as a useful research tool rather than simply an educational or teaching aid. Scientists have long been able to work collaboratively at a distance; however, it is often in terms of receiving data or sending some instructions where there may be a delay in receipt of the information. When defining remote control it is important to note that electron microscopy requires instantaneous control and receipt of the feedback (in most cases via images on a screen). Without realtime control it is impossible to conduct high resolution imaging and analysis work. In terms of electron microscopy, there are several reasons for conducting experiments remotely: With sub-Ångström aberration-corrected scanning transmission electron microscopes, the environment within which the microscope itself sits is of utmost importance.

Microscopists have been identifying particulate matter since the seventeenth century. Reference sets of study slides, identification keys, and even atlases of specific groups of microscopic substances were prepared throughout the eighteenth and nineteenth centuries. During the latter half of the nineteenth century and the early twentieth century, these types of resources grew in volume, but none of them attempted to be a comprehensive source.

In 1967, Dr. Walter C. McCrone and his colleagues changed the practice of microscopy with the publication of The Particle Atlas, Edition I. This single volume first edition, a photomicrographic atlas, illustrated and described 404 substances based on analyses using the polarized light microscope.

Zoysia, a common turf grass, is characterized by the presence of functional salt glands. These glands are specialized structures through which the plants excrete excess salt. Research on the mechanism of salt secretion in Zoysia matrella (Manila grass) prompted the development of a specimen preparation technique that would preserve the secreted salt and salt gland. Conventional aqueous preparative techniques wash away the secreted salt on the leaf surface. A specimen preparation technique was modified from a simple cryo-preparative technique for examining hydrogels in the transmission electron microscope.

2D-crystallization in combination with transmission electron microscopy (TEM) is one of the few methods for the structural analysis of membrane proteins in their native state. However, the parameters for the generation of large crystalline sheets are typically difficult to identify for a given protein. Many repetitive and time consuming screening steps by TEM are therefore necessary to find the best crystallization and preparation conditions. Although several software packages offer the possibility to control an electron microscope, none is completely adapted for a fully automated and completely integrated acquisition and analysis of 2D crystals. Here we report on the development of a fully automatic screening and on-line analysis software for the fast and automatic survey of large quantities of negatively stained EM samples for 2D crystallography.

Most spectroscopies (FTIR, XRD, RAMAN, MS, etc.), were developed in order to identify materials. This is accomplished by capturing the spectral “signature” of an unknown and matching it to those of a comprehensive database of reference materials. Development of these spectroscopies has included efforts to embellish the associated database. EDS evolved as a resource to provide qual/quant, and development has been to improve the accuracy of quantation. Until recently, EDS was not considered a spectroscopy to provide identification of materials.

As you all know, recharge: the price or rate a microscopy facility charges others to use their instruments, is a big topic on the MSA listserver and everybody struggles with designing rate systems. My talk is geared to U.S. labs and a lot of our restrictions regarding rates are not the same in Canada or other countries around the world. Our goal is designing recharge systems that will encourage facility use that will also recover the cost of the operation. How much you have to recover depends on your particular operation, your administration, and institutional requirements.

Electron backscattering diffraction (EBSD) has become a technique of great importance in scanning electron microscopy. The diffraction patterns obtained by this method are of value in identifying phases and in determining the orientation of samples. The technique was initially developed about thirty years ago and has enjoyed widespread use in the last decade or so. It is therefore something of a surprise that there do not exist computer programs that simulate the patterns well. In fairness, we must make it clear that a number of programs exist and others are being implemented, which represent important steps towards the simulation of EBSD patterns. However, as we hope these notes will make clear, a satisfactory simulation will be very difficult and we are not aware of any existing or forthcoming program that does a job that can be guaranteed to give accurate values for the intensities in the experimental patterns.

Biological materials interest biologists and engineers for their complex interactions among constituents and unique mechanical properties. While the heterogeneous tissue structure and its material properties are responsible for intriguing biomechanics, they pose challenges for sectioning, particularly in regions with stark tissue boundaries. Microscopists level this playing field when sectioning samples by embedding them in paraffin or plastic; but for scanning electron microscopy, natural morphology must be preserved without sacrificing the sample’s surface contours. Here we outline a simple preparation method for visualizing in cryoSEM the calcified cartilage of sharks and rays (elasmobranch fishes), a layered biocomposite that has traditionally been considered difficult to prepare for microscopy.

The fact that the machinery is “behind the wall” or out of sight may mean that it is not being looked after as well as it should. Do you have a regular schedule to check the vacuum pumps and accessories in your microscopy lab, or do you wait till it fails? Do you keep a service record of the repairs done to your vacuum pumps and the vacuum levels measured after each service? In a department where there are instruments that last ten years or more and the operators and technicians regularly change it makes good sense to keep a record of the vacuum pumps oil changes, service and performance.

Cryo-fracture, in conjunction with critical point drying is a method used to prepare biological samples in order to expose, for viewing via scanning electron microscopy, those naturally occurring surfaces which might otherwise remain obscure. For example, the Bowman’s capsule and tubules of a kidney, tiny blood vessels on any organ, inter-cellular spaces in liver or alveoli in the lungs. Also, some surfaces, not normally exposed at all such as the membrane surface of a nuclear envelope, mitochondria or chloroplasts or the cytoplasm of a cell, can be brought to light with this method. Herein is a review of the development of cryo-fracture and how it is currently used at our facility.

A purpose of microscopy is to magnify and enhance contrast between different regions of a sample, whether those regions may be different structures, different orientations of the same structure, regions of different atomic weight, or different chemistries. In the present case, elemental mapping in the energy filtered transmission electron microscope (EFTEM) is used to enhance contrast between elements in an apparent bundle of fibers previously seen through scanning electron microscopy (SEM) and a particle that appears to be a catalytic source for the fibers.

Remote control of a scientific instrument is a topic gaining more and more attention between instrument users and operators. The project presented in this article reports results obtained from two distinct research efforts. The main outcome from the first research was the realization of an application to remote-control a Scanning Electron Microscope (SEM), while the main outcome from the second research was the implementation of a procedure to reconstruct 3D surfaces.

Poly-L-lysine of a medium molecular weight, 30,000 to 70,000 Daltons (Sigma–Aldrich), is currently used in our facility as a polycationic adhesive on 5 mm square silicon chips to secure individual anionic cells and particles for ease of processing and viewing in the scanning electron microscope. In this technique, a solution of 1 mg/ml of poly-L-lysine is dissolved in distilled water. Drops of the solution are placed onto clean 5 mm square silicon chips and allowed to sit for one to several hours before being wicked away. The prepared chips are used immediately. Suspended cells are then applied to the chips and allowed to settle and adhere, after which they are washed and fixed. Alternatively, fixed suspended cells or particles are applied to these silicon chips. The cells and particles secured to the silicon chips are easily handled through processing steps such as dehydration, critical point drying, and metal sputter coating for viewing in the scanning electron microscope. Presented herein is a brief history of the evolution of the technique of using poly-L-lysine as an adhesive for SEM.