Microwave processing techniques are improving at an incredible rate. We use microwave processing for fluorescence labeling, immunogold labeling, decalcification, antigen retrieval, and TEM and SEM processing for biological specimens. If it works conventionally, it is likely to work in the microwave. The advantage is savings in chemicals, time, and labor. Typically, fluorescence labeling, with the cells mounted on slides ready for the confocal microscope, takes 30 minutes. The cells are fresher and there is usually less background. For TEM processing, the time from fixing to cutting is 2-3 hours. Microwave processing comes between high pressure freezing and conventional processing in regard to the amount of extraction that takes place. This talk will explain the workings of the microwave and give examples of microwave results in comparison with conventional techniques.

Cryo-electron tomography is a rapidly developing technique that produces 3D images of frozen-hydrated specimens with much potential for the characterization of biological structures and macromolecular assemblies in situ. Such information can provide detailed insights into the structural basis and ultimately the function of many cellular processes. A general introduction into the fundamentals of cryo-preparation and electron tomography will be given. We will also demonstrate and discuss both strengths and limitations of the technique in theory and with cell biological examples.

The availability of 3-D microstructural information is becoming increasingly important for understanding the complex microstructure-property relationships in advanced materials. Mechanical serial sectioning is a well-established technique for obtaining 3-D microstructural data from standard metallographic specimens. Traditionally, layer removal has been done by hand, using standard metallographic polishing techniques. This tutorial will begin with an introduction to manual serial-sectioning methods, their applicability and limitations. Special emphasis will be given to techniques for maximizing the fidelity of the data, including imaging and alignment issues and useful image processing tools. The tutorial then outlines modern methods for automation including robotic specimen manipulation, automatic image capture and automatic specimen prep which can reduce the time to acquire data volumes by around 2 orders of magnitude, compared with manual techniques. Selected examples will be given where automation has enabled the systematic study of microstructures, using a variety of experimental set-ups. The tutorial also will discuss the future of the technique.

Acquiring 3D data sets by means of serial sectioning techniques requires careful alignment of the individual 2D images. In this tutorial, we will work through the various image processing steps that need to be carried out in order to convert a 2D image stack into a 3D object. We will discuss cross-correlation techniques, histogram adjustments, and a variety of other approaches. A brief review of various free and commercial software packages will also be included.

The ability to characterize material microstructure in 3-D is a critical methodology for developing unbiased structure-property relationships. This tutorial will cover experimental approaches to use and optimize the Dual-Beam Focused Ion Beam Scanning Electron Microscope (DB FIB-SEM) to perform serial sectioning experiments at the micron-size scale. The tutorial will cover the state-of-the-art advances as well as current limitations of this technique, and in particular will cover the integration of ion beam imaging, EBSD and EDS mapping into the serial sectioning experiment.

A three-part tutorial program will be compiled to help microscopists and materials scientists (1) understand the state-of-the-art in instrumentation for atom probe tomography and, (2) learn about advanced specimen preparation techniques. And (3) explore applications in the study of advanced nanomaterials. This will include studies of Si-based semiconductor devices, advanced aluminium alloys, zirconium alloys, and spinodal systems and involve an analysis of solute distribution, states of clustering, nucleation and the characterisation of nanoscale precipitation and phase decomposition, beam imaging, EBSD and EDS mapping into the serial sectioning experiment.

This tutorial covers many aspects of CCD technology as they relate to modern TEM functionality. Fundamentals of CCD camera construction, including scintillator considerations, number of pixels, pixel size, camera speed versus read-out speed, lens coupled versus fiber optic coupled, and speed versus sensitivity trade-offs will be discussed as they relate to the actual use of the TEM. Use of TV cameras on a TEM will also be addressed. By the end of the tutorial, the attendee will be able to make a better informed decision as to what moving up to a digital camera will bring about in terms of benefits to the users EM facility. Potential future directions in this developing area will also be discussed. Both the materials sciences and the life sciences TEM user will be addressed.

Electron tomography is a powerful method for obtaining the three-dimensional structure of a specimen from a set of images taken at a series of tilt angles. This tutorial will introduce the basic principles of tomography, then go through the series of steps involved in building a tomographic reconstruction with the IMOD software package. This will include the reconstruction of a single volume from tilt series taken around two orthogonal axes, a method that can significantly improve results for some specimens. The tutorial will also cover how to deal with some of the problems commonly encountered when doing tomographic reconstructions.

The successful "Experts" sessions began in 1997 as a novel means of communicating scientific knowledge. Essentially, hot topics are identified and experts are located to answer questions posed from the audience. One or more invited "experts" makes a short presentation to introduce the topic. Audience members then ask questions and the "experts" provide answers. Other members of the audience may also contribute answers to some problems. Sharing of experience and information is strongly encouraged as the purpose is to have a dynamic discussion among the panel and audience participants. The topics for 2006 are:

Physical Sciences
Routine and Advanced Applications of EBSD
Organizer: Roy Geiss
Experts will describe both routine and advanced applications of electron backscatter diffraction in material science. Discussion topics will include orientation imaging, phase identification and strain measurement.

Biological Sciences
Application of Low Voltage STEM
Organizer: Edward Principe

Experts will describe the applications of low voltage STEM imaging in the biological as well as material science disciplines. Discussion topics will include advantages, disadvantages and limitations of low voltage STEM as well as possible future directions for the technology.