Whereas too much of a good thing can be bad, too little of a good tiling can be good. In this case, the accidental dilution of X-rhodamine tubulin injected into living cells resulted in a heterogeneous labeling of micro tubules, rather than visualization of continuous structures. This heterogeneous labeling rendered a speckled image, hence the term fluorescent speckle microscopy (FSM) was introduced. It turns out that FSM yields particularly useful information on dynamic events within living cells, for example the assembly and disassembly of microtubules.

There are two recent reviews of FSM by Clare Waterman-Storer and Gaudenz Danuser, one emphasizing the biologic applications of the technique, the other emphasizing the quantitative aspects. For their purposes, they defined a “speckle” as a diffraction-limited region of the image that is significantly brighter than its immediate environment. It can be calculated from the point-spread function (which is determined by the numerical aperture of the lens) that optimally a diffraction-limited region of about 250 nm can be imaged.

Coal is remnants of plant matter that were compressed and held at a high temperature for eons. However, if plants are charred first, they become hard and brittle, and may resist compression. Under appropriate circumstances, anatomic detail can be elegantly retained. Ian Glasspool, Dianne Edwards, and Lindsey Axe have performed detailed studies on plant fossils recovered in England, near the Welsh border, that can be considered to be evidence for the earliest wildfire yet described on our planet. They examined a series of mesofossils, about one millimeter in size, which required microscopic study.

It is well known that chemical and structural changes occur to organic tissues during charring. This can mean that charcoal is resistant to decay and compression and it is also more reflective than non-charred material. Glasspool et al. used incident light and scanning electron microscopy to reveal 3-dimensional cellular detail of the specimens. Additional specimens were embedded and polished for reflectance microscopy. The results indicated that the specimens were charcoal.

Intracellular communication is imperative for multicellular organisms. Such devices as synapses and gap junctions have been recognized for decades. Now Amin Rustom, Raiser Saffrich, Ivanka Markovic, Paul Walther, and Hans-Hermann Gerdes have described a new model of cell-to-cell communication.

While looking at PC12 (rat pheochromocytoma) cells in the presence of fluorescently labeled wheat germ agglutinin, Rustom et al. observed relatively long connections extending between cells. These structures were 50 to 200 nm in diameter and up to several cell diameters in length and were named tunneling nanotubes (TNTs). TNTs were subsequently found connecting cultured cells from other lines. They were consistently positioned along the smallest distance between the cells, did not contact the substrate, and occasionally were branched. TNTs immunostained positive for actin, but did not contain microtubules. Scanning and transmission electron microscopy definitively established that a TNT represented a seamless continuity of the plasma membrane from one cell to another.

They're right at your fingertips, but how often do you consider the microscopic anatomy of your fingernails? How is their form fitted to function? Primates, including humans, use fingernails to maintain the anatomic integrity at the distal end of our phalanges, which often serves as the leading edge of our physical sensory apparatus. The nails serve as a stiff backing to the soft pads of our fingers, helping to improve the grip and sensitivity by allowing the pad to “push back” on the object we are touching. Nails also serve specific functions in maneuvering within cracks, scratching, and fighting.

The gross structure of nails helps to resist upward bending forces that are encountered in these functions. The convex curvature contributes, just like a curved piece of paper has more stiffness than a flat sheet. The composition of nails is also suitable. As with the outer layer of skin, and the structure of hair, nails are also composed of keratin.

Would it be useful if you could examine a wet, perhaps even a living, specimen in the scanning electron microscope? Of course we think that this would be impossible, given the vacuum the specimen would be subjected to in the microscope. However, in the realm of materials research, the Environmental Scanning Electron Microscope (ESEM) is being developed. Apparently this instrument has not been fully appreciated and utilized by biologists.

As reviewed by Athene Donald, a pioneer in developing the ESEM, this instrument has been commercially available for a decade. It is still evolving as a useful tool in looking at materials such as cement, natural fibers, and aqueous dispersions. Applications of the ESEM. to biological Studies are extremely limited. However, studies on biological samples, even living cells, are possible (although not without problems and limitations).

The use of fluorescent probes is becoming more and more common in cell biology. It would be useful if we were able to correlate a fluorescent structure with an electron microscopic image. The ability to definitively identify a fluorescent organelle would be very valuable. Recently, Ying Ren, Michael Kruhlak, and David Bazett-Jones devised a clever technique to correlate a structure visualized in the light microscope, even a fluorescing cell, with transmission electron microscopy (TEM).

Two keys to the technique of Ren et al are the use of grids (as used in the TEM) with widely spaced grid bars and the use of Quetol as the embedding resin. The grids allow for cells to be identified between the grid bars, and in turn the bars are used to keep the cell of interest in register throughout the processing for TEM. Quetol resin was used for embedding because of its low auto fluorescence and sectioning properties. The resin also becomes soft and can be cut and easily peeled from glass coverslips when heated to 70°C.

Once in a while we have the privilege of participating in an activity that promises to radically improve our technical capability to explore materials at the microscopic level. The recent practical demonstration of aberration correction in the SEM, the TEM and the STEM signify nothing less than a revolution in the way that we design and operate electron optics for microscopy.

Scherzer understood from the beginning of electron microscopy that an open magnetic lens having cylindrical symmetry will have positive, non-zero aberration coefficients, and so therefore will always have some minimum aberrations. In practical instruments, it was arranged that the limiting defect would be spherical aberration in the objective lens. This allowed the imaging performance to be largely characterized by a single parameter, spherical aberration, and also allowed a certain amount of image simulation and post-experiment analysis to facilitate interpretation of imaging results.

I am a botanist, with an expertise in plant anatomy, specifically wood anatomy. In my capacity of mkroscopist for the Smithsonian Institution, I help answer research questions developed by curators, conservators, anthropologists and archaeologists. I am typically presented with a variety of materials that range from large, entire objects from museums to small, deteriorated, fragmentary samples from archaeological digs. Many of the requests center around the questions “What is it?” and “Where did it come from?” The answers to these questions help determine the traditional materials used by a culture and the distribution of these objects through trade and cultural exchange. Of the numerous requests that I receive, one or two each year tend to stand out as captivating examples of applied optical microscopy. The one that I would like to present in this article (along with my intern for the project - Alvaro Galvis) is the early Catholic statues (santos) from Chihuahua, Mexico and southern California.

The Materials Science Program of NASA's Office of Biological and Physical Research (OB PR) has attacked futidamental problems of materials science as defined by an external advisory group, known as the Discipline Working Group (DWG). These have been:

1. • Nucleation and Metastable States

2. • Prediction and Gontrol of Micrestructures

3. • Crystal Growth and Detect Generation

4. • Phase Separation and Interfacial Phenomena

5. • Thermophysical Properties and Process Modeling.

While the program's primary objectives are science-based and despite the fact that 90% of these programs concentrate on pre-cursor and theoretical research on the ground, NASA demands that there be a microgravity rationale driving the research.

Historically near-infrared (NIR) spectroscopy has enjoyed significant success as the industrial ‘cousin’ of the mid-infrared FTIR spectrometer. Most NIR spectrometers are significantly more rugged than their MIR counterparts, and are therefore suited for use as a QA/QC tool. Sample preparation is trivial or non-existent for most solid-state or liquid samples and data collection is rapid, while still achieving excellent signal to noise. All these factors combine to produce an analytical measurement technology with widespread applicability. Until relatively recently, its capabilities as a microspectroscopy and chemical imaging tool has been largely underutilized. Most NIR spectra have been collected from bulk, not microspectroscopic samples.

Mid-infrared (MIR) microscopy, based on a Fourier Transform infrared (FTIR) spectrometer, has been the infrared microspectroscopy technique most readily available up to now. It is a broadly employed chemical identification tool that is used to complement traditional light microscopy.

The small unassuming nematode, Caenorhabditis elegans is only one millimeter long and lives in the soil munching on bacteria. While many nematode (roundworm) species are parasites with medical or agricultural importance, C. elegans seems to harm no one. Yet, this animal has attained a status in medical science that compares to more complex organisms such as the mouse or fruit fly in its utility for scientific discovery. It has been the subject of thousands of studies dealing with topics as diverse as nutrition, aging, and nervous system development. About 5000 scientists are now pursuing this single species in hundreds of laboratories worldwide. In 2002, the Nobel Prize in Medicine was awarded to three of the pioneers in establishing C. elegans as a “model organism“: Sydney Brenner, John Sulston, and H. Robert Horvitz. Why study worms?

Sydney Brenner first turned his attention to C. elegans in the 1960's. Working at the Medical Research Council in England, he was looking for a small animal with inexpensive tastes that could be easily cultured in the laboratory.

Since publication of the classic text on the electron microscope laboratory by Anderson, the proliferation of microscopes with field emission guns, imaging filters and hardware spherical aberration correctors (giving higher spatial and energy resolution) has resulted in the need to construct special laboratories. As resolutions iinprovel transmission electron microscopes (TEMs) and scanning transmission electron microscopes (STEMs) become more sensitive to ambient conditions. State-of-the-art electron microscopes require state-of-the-art environments, and this means careful design and implementation of microscope sites, from the microscope room to the building that surrounds it. Laboratories have been constructed to house high-sensitive instruments with resolutions ranging down to sub-Angstrom levels; we present the various design philosophies used for some of these laboratories and our experiences with them. Four facilities are described: the National Center for Electron Microscopy OAM Laboratory at LBNL; the FEGTEM Facility at the University of Sheffield; the Center for Integrative Molecular Biosciences at TSRI; and the Advanced Microscopy Laboratory at ORNL.

In a departure from traditional articles on Photoshop, this article will provide a look at how to save movies for the purpose of distribution to Macintosh, Windows and Linux operating systems.

More than once, I have been asked to produce and help with the distribution of digital videos that rival those downloadable trailers for big Hollywood productions. I kindly inform the clients that I do not have a staff large enough to tweak each individual frame of their video, and that their expectation must go down a few notches. Upon seeing their baffled look, an explanation ensues, much like what follows:

First of all, digital video must be compressed in order to run seamlessly from a computer. Only the most robust computers can keep up with the number of frames flashing by per second (-30 frames for NTSC), and even these cannot play back for extended periods of time without the loss of some frames (what is called dropped frames in the digital editing world).

Microscopy is one of the most important techniques in microbial ecology, since this is the most direct approach to examine the microbe's world from its own perspective and spatial scale. The value of quantitative microscopy in studies of microbial ecology can be increased even further when used in conjunction with computer assisted image analysis. There are two main advantages of using digital image processing and pattern recognition techniques in conjunction with microscopy for quantitative studies of microbial ecology. First, automatic image analysis reduces the amount of tedious work with microscopes needed to accurately quantify in situ morphological diversity, abundance and metabolic activity of microbes. Secondly, these techniques provide an important quantitative tool that can significantly enhance the polyphasic analysis of the structure, diversity, spatial features, and functions of complex microbial communities in situ without cultivation.

The invention of scanning tunneling microscopy (STM) in 1982 revolutionized surface analysis by providing atomic-scale surface imaging of conducting and semiconducting materials. Shortly after that, atomic force microscopy (AFM) was introduced as an accessory of STM for high-resolution imaging of surfaces independent of their conductivity. Mechanical force interactions between a sharp tip placed at one end of a micro fabricated cantilever and a sample surface were employed for imaging in this method. In the past decade, AFM has developed into a leading scanning probe technique applied in many fields of fundamental and industrial research. The progress of AFM has been made possible by implementation of an optical level detection scheme, which allows precise measuring of the cantilever deflection caused by the tip-sample forces, by mass microfabrication of probes consisting of cantilevers, and by developments of oscillatory imaging modes, particularly, Tapping ModeTM.

The high cost of Photoshop and its increasing complexity with each new version can leave the user longing for a version that is a fraction of the price, but still contains the basic features. That longing may be met with Adobe's Photoshop Elements.

Certainly its price is attractive at $99.00 on some websites and about $50.00 on others (including sites for those in Education), For the shopper who wishes to do a little extra research, a web search may reveal that Elements is included with a scanner or ink jet printer at well under $99.00, making the purchase a 2-for-1.

Elements is a pared down version of Photoshop, but it is still a viable solution for almost all image processing needs. Simply because it is pared down, it becomes very attractive for occasional users who do not desire to re-learn the full version of Photoshop, let alone yet another upgrade and yet more frustrating minutea to discover.

The newest generation of computer-controlled electron microscopes incorporates the ability to perform adjustments to microscopy conditions by comparing pairs of images and altering the conditions accordingly. Automation of electron microscope adjustments offers the advantages of accuracy, precision, efficiency, the ability to incorporate the adjustments into other automated procedures, and, for radiation-sensitive specimens, minimal exposure to the beam.

At present, automated functions include determination of eucentric height, focus, astigmatism, orientation and location of the stage tilt axis, centering of an image feature, and rotation center alignment. It is possible to automate other functions, so this list may be incomplete. In general, these functions are accomplished by induced image shifts, and, in many cases, the automated functions are completely analogous to the corresponding manual ones.

The use of microscopy and imaging methods to study foods is not a new idea. It has been going on since the first light microscopes were developed (White, 1970; White and Shenton, 1974-1984). Microscopy has been used to determine the quality, purity and safety of foodstuffs by detecting and identifying contaminants in foods. The short article by Stephen Carmichael in the May/June 2002 issue of Microscopy Today has again brought food microscopy into the spotlight. The article provided an opportunity to discuss present applications of food microscopy and to give some projections of where it is headed in the future. The reader may not realize that microscopy and imaging methods are used extensively by most major food companies worldwide for product development, quality control, and trouble shooting (Allan-Wojtas, 1999). Often, this work cannot be published because it contains proprietary information. The application of microscopy to food structure analysis is one of the best kept secrets in microscopy today.

PowerPoint has traditionally been the software of choice for slide shows tit conferences and meetings. Its ease of use has made the program ideal, along with its universality among scientists and engineers.

The program does have its drawbacks, however. Most notably, it is saved in a format that essentially makes it unusable for any other purpose than that of a slide show. It cannot be used for publication, and, as many discover all too late, it is not the software of choice for making posters (both these phenomena go back to Microsoft's refusal to pay a license to Adobe for PostScript, the universally used “page description language” among publishers). The remainder of the drawbacks exist when the slide show needs to be distributed. Newer versions of PowerPoint can make HTML files for posting on a web site by using Save As under File, but saving in the PowerPoint format has not evolved as a way to view slide shows on the web.

Several images can be stitched together (sometimes referred to as “montaging”) through the use of Photoshop. Edges of the images can be feathered and the process automated to reduce the hours of work that can go into this procedure.

Before going any further, let me be quick to say that more than one company has developed protocols to accomplish image stitching. When your lab is looking at large fields, while needing to resolve fine details, then I would strongly recommend the purchase of a system designed for this purpose. Issues surrounding motorized stage accuracy, problems with tilting when a camera is perched on a long optical tube, and image to image correction for contrast, color and uneven illumination can all contribute to confounding issues when image stitching. These companies have worked through these problems with off-the-shelf systems.