Pre-meeting Sunday Short Courses: August 5
Organizers: John Mansfield and Louis Kerr

21-01: Practical Digital Imaging
Full Day: 9:00 AM - 5:00 PM
Instructor: John MacKenzie

This course will discuss the various strategies needed for producing digital data that is suitable for publication. We will discuss how to acquire the best digital image for a given sample. Noise removal and resolution issues will be discussed in detail. We will examine what the current best technologies for archiving the image data are and what image formats and standards we should adopt. We will examine in detail image printing. We will emphasize several issues that must be understood in order to produce high quality images every time on any printer (the most critical being the gamma correction). There will be a strong emphasis placed on the most affordable solutions available regardless of platform or operating system. We will examine the latest technologies such as digital cameras and digital video to see how they may best be applied to microscopy. We will discuss the major issues that must be addressed when moving to a more digital approach.

21-02: Image Processing and Analysis
Full Day: 9:00 AM - 5:00 PM
Instructor: John Russ

This will be an intensive step-by-step illustration of the various steps involved in enhancing images for presentation and extracting numeric data from them for analysis. The emphasis will be on comparison between various approaches applied to representative images, rather than the theoretical underpinnings for the various techniques.

Morning: Survey of the principal techniques for image processing will cover spatial domain operations such as histogram modification, convolution with filters, neighborhood ranking operations, etc., showing their use for removal of random noise, correction of nonuniform brightness, enhancement of edges and local detail, etc. Fourier domain processing will be used to remove periodic noise, deconvolve image blur, locate features by cross correlation, and isolate periodic structures.

Afternoon: Thresholding of images, and processing of the binary images using morphological operations such as erosion and dilation, skeletonization, watershed segmentation, etc., will be used to delineate features of interest for measurement. The Boolean combination of images and the use of appropriate grids allows straightforward stereological measurements of 3D structure to be performed. Feature-specific measurements provide data on object density (or color), position, size and shape.

21-03: Low Voltage and Low Vacuum SEM
Full Day: 9:00 AM - 5:00 PM
Instructors: David Joy and Oliver Wells

Present day scanning electron microscopy is dominated by the need to achieve high-resolution images of surfaces, by the problem of imaging materials which are insulators or poor conductors, and by the desire to combine imaging with chemical microanalysis. This short course will focus on two different approaches to satisfying these needs - Low Voltage Microscopy and Low Vacuum Microscopy.

A low energy electron beam has limited penetration into a solid and so produces more surface specific imaging information than a conventional higher energy beam. The mechanisms of formation and the interpretation of low voltage images will be discussed and illustrated and the associated problems of sample cleanliness and beam-induced damage will also be considered. A variety of practical techniques to obtain stable images from non-conducting materials in the low voltage SEM, including charge balance, dynamic charge control, detector strategy, and coatings, will be described and evaluated. The use of X-ray microanalysis at low beam energies will be discussed and the benefits and pitfalls of microanalysis at low incident energies will be identified.

When gas is introduced into an SEM the interaction of the electron beam with the surface is significantly modified. A wider range of materials can be examined, including those that are damp or dirty or wet, new image contrast mechanisms become available, and charging can be suppressed even for the most difficult samples. The basic principles of Environmental and Variable Pressure SEMs will be presented and the novel aspects of their operation will be discussed. The effect of the gas on the imaging and analytical resolution and on the operations of the detectors will be considered. Finally some of the wide ranging applications of this new technology will be illustrated.

21-04: What the Heck Happened to This?? - Real Life Case Studies of Failure Analysis
Using Microscopy and Microanalysis.
Full Day 9:00AM - 5:00PM
Instructor: Valerie Woodward

Most industrial service lab microscopists are faced with solving manufacturing and applied R&D problems on the fly with broad-use, commercially available, and sometimes outdated equipment using routine analytical methods. Although the questions about why materials fail (or work!) aren't always answered at the most fundamental levels, we do need to provide the best reasonable answers in the shortest reasonable times to our "customers" so that they can relate the problem to a specific process or material. Many times, a combination of analytical methods, including outsourcing work to labs that have the needed instrumentation, is necessary, and the microscopist has to be well versed in those methods, and the materials and the processes, in order to determine what analyses are needed and to pull together all of the results in order to solve the problem. This course will present some approaches to failure analysis, and a number of case studies that required the use of multiple hierarchies of microscopy, microanalysis, microsampling, and multiple analytical techniques in order to provide timely and useful results to the customers.

21-05: Materials Ultramicrotomy: Polymers and Hard Materials
Full Day: 9:00 AM - 5:00 PM
Instructor: Helmut Gnaegi

Ultramicrotomy has been used for a long time to prepare biological samples for TEM. In the field of materials science, this technique gains increasing attention. For the preparation of ultrathin sections of polymers, (cryo) ultramicrotomy with diamond knives is well accepted. However, ultramicrotomy is seldom used for the sectioning of hard industrial materials, probably because of the brittleness of the samples and of possible damage caused to the diamond knives. Recent publications have shown that even the hardest known materials may be sectioned successfully, if the important preparation steps such as sample trimming, embedding, etc, are done with great care. Ultramicrotomy of materials has thus proved to be a fast and efficient technique not only for TEM, but also for surfacing samples for SEM, STM and AFM. The course will cover how to prepare unknown polymer samples, including: mounting samples, trimming, room temperature sectioning, cryo-sectioning, staining (block staining, section staining, OsO4 and RuO4 staining, double staining) and safety issues, etc. Hard materials will also be featured with an introduction, sample primer treatment, embedding, trimming and sectioning, section collection and mounting on grids. Live demonstrations of polymeric trimming and sectioning (room and low temperature) and section collection and mounting on grids will follow the lecture section of this course. Participants are invited to bring their own samples. The preparation steps for each sample will be discussed. If time allows, participant samples may be sectioned at the end of the day.