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July 31 |
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SC01: Towards Nanoscale Imaging of Anything in VPSEM (including ESEM): From Basics to Current Practices |
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Brendan Griffin is the Director of a multi-user facility (200+ users) and was an original ESEM developer in 1990. His laboratory's mission is to facilitate access and usage by researchers in current microanalytical technologies and their VPSEM is used for ~3000hrs annually with ~140 new SEM users trained annually. This course will address the application of VPSEM and the extended pressure range (ESEM type) instruments in two sections. The course will open with a tutorial style review of the current variable pressure and environmental SEM instrumentation with specific attention to the variety of gaseous or variable pressure secondary electron (VPSE) detectors. The benefits of various electron sources will then be considered, together with a discussion of samples suitable for image quality control protocols. The second core area for discussion is the electron-gas interactions and their roles in image production. The influence of the different operating conditions will be considered in detail, using theoretical and practical examples. The benefits, and otherwise, of the range of potential imaging gases will also be compared. X-ray microanalysis procedures, dynamic experimentation and extended pressure operation will be described and particular attention will be paid to the imaging of fully hydrated samples. All topics will be supported with images and, in each segment, open discussion will be encouraged. Following the very successful forum in 2004, relevant manufacturers will separately present cameos of their current and future visions prior to a final round-table general discussion. Attendees of this course will then be able to attend the VPSEM Symposium in the M&M 2005 main meeting to learn about the leading edge advances in the technique. |
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SC02: Image Processing and Analysis |
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John Russ is the author of The Image Processing Handbook, Computer Assisted Microscopy, Practical Stereology, and Forensic Uses of Digital Imaging, as well more than 300 papers and 12 books. He has been involved in the use of a wide variety of microscopy techniques and the computerized analysis of microstructural images for nearly 50 years. He is a retired professor from the Materials Science Department at North Carolina State University, where he continues to write and lecture on topics related to image analysis. This will be an intensive step-bystep 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: A 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. |
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SC03: Photoshop for Microscopy and Microanalysis |
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John Mackenzie runs the Center for Electron Microscopy at North Carolina State University that services twenty-two Departments in the College of Agriculture and Life Sciences. His research interests include instrument design and development with an emphasis on computer control and digital imaging. For the last ten years he has taught courses in digital imaging at both the national and international level. The workflow for digital imaging in the modern microscopy and microanalysis laboratory is still a work in progress. This new course offering will use Adobe Photoshop as the core program in defining this workflow. We will show how digital images can be properly manipulated in Photoshop to present the best image for display, reports, and publication. The course will use real world images to demonstrate the basic Photoshop manipulations of histogram stretching and gamma adjustment in a step-by-step manner. There will be an emphasis on how this approach solves most of the problems encountered in scientific imaging. We will see why Photoshop layers can be a powerful tool in labeling and making composite images. This step-by-step lesson will show the ease and flexibility of this approach. We will discuss the issues of file formats, compression, and ethics so that the proper documentation of our manipulations is kept. Several of the new features in Photoshop CS (Photoshop 8.0) will be explored in detail. These features build on the introductory lessons and show how the use of adjustment layers, history logs, and XMP file format extensions can insure total control of an image's history. Finally, archival image storage options will be addressed. |
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SC04: High Pressure Freezing, Cryosectioning of Vitrified Samples for Tomography, and Freeze Substitution |
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Kent McDonald has been practicing electron microscopy for over 35 years, and for the past 15 has been using high pressure freezing and other cryotechniques to investigate problems in cell biology. He is currently the Director of the Electron Microscope Laboratory at the University of California, Berkeley. After finishing his PhD in 1996 in the Lab of Arie Verkleij, Utrecht, the Netherlands, Verkade then worked at Kai Simons' lab at the EMBL in Heidelberg, Germany. In 2001, he joined the Max Planck Institute of Molecular Cell Biology and Genetics in Dresden, Germany to set up the electron microscopy department, where he is now focusing on applying new techniques in the field of intracellular transport. Helmut Gnaegi has been with Diatome since 1970, where he developed the technique for making diamond knives for ultramicrotomy. He has taught many ultramicrotomy courses and seminars in biology and materials sciences in Europe, the U.S. and the Asia Pacific. New techniques and accessories for high pressure freezing, freeze substitution and cryosectioning of biological samples are evolving rapidly.Accordingly, this year's course will emphasize the practical aspects of this technology such as how to optimize high pressure freezing for a wide variety of samples, the best strategies for freeze substitution, and how to cut cryosections of frozen hydrated as well as sucrose-infiltrated material. Using a "live" video format, participants will see step-by-step procedures for loading samples for high pressure freezing, including many that are new or unpublished. Current models of high-pressure freezers and their specimen holders will be considered in detail. Following high pressure freezing, some samples will be transferred to a cryoultramicrotome and techniques for cutting frozen hydrated cryosections will be demonstrated. Ample time will also be allotted for roundtable discussions of freeze substitution and other low temperature methods. Persons taking this course should leave with a working knowledge of these cryotechniques that they can apply immediately to their own research, whether it involves tomography, cryosectioning, EM immunolabeling, or they just want to have the best available preservation of cellular fine structure. |
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SC05: Specimen Preparation for the Physical Sciences |
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Before joining PPG, Scott Walck worked with the Tribology and Surface Interactions groups characterizing thin films by SEM and TEM. He joined PPG as a Team Leader for the Materials Characterization Group where he worked on both production and research thin film projects. He is currently a project leader in the Flat Glass New Products and Processes Group developing and improving thin films on glass. Ron Anderson is presently editor of Microscopy Today magazine. Ron recently retired from IBM where he performed TEM analysis on semiconductor and ceramic specimens since shortly after the middle of the last century. While at IBM, Ron and his coworkers developed numerous specimen preparation tools and protocols. His laboratory is best known for the invention of the tripod polisher. This course will cover both wellknown and innovative techniques for the preparation of plan view and cross sectional TEM samples for physical science applications. Well established recipes will be covered as well as guidelines for students developing their own sample-specific procedures. Specific topics presented will include mechanical pre-thinning, electro-polishing, dimpling, tripod polishing, ion milling, focused ion beam (FIB) milling, ultramicrotomy, cleavage, and chemical polishing. Students will be made aware of the root causes of the creation of artifacts in samples prepared by these methods, and protocols for artifact recognition, control, and minimization. The instructors have experience with a wide variety of materials and active participation by the students with respect to their experiences and problems will be encouraged. Students at all levels of experience in TEM can benefit from this course. |
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SC06: Digital Microscopy and
Image Analysis for Materials
Characterization |
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Sidnei Paciornik is Associate Professor at the Department of Materials Science and Metallurgy of the Catholic University of Rio de Janeiro. For the past 15 years, he has been working with image analysis for materials characterization and is currently integrating microscope automation and image analysis, creating new methods for materials characterization. In recent years, the integration between the microscope and computer has been growing steadily. Digital image acquisition, motorized, computer-controlled microscopes and powerful image processing and analysis software has opened the doors to novel micro-characterization techniques. This course will introduce the main developments in these techniques, and show their impact in materials characterization. The classical sequence of image acquisition, pre-processing, segmentation, postprocessing and feature extraction will be presented and linked to microscope automation. Fully automatic materials characterization will be demonstrated with routines such as auto-focus, extended focus, and extended field among others. |
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SC07: Interpretation of Microstructure |
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George F. Vander Voort is Director, Research & Technology at Buehler Ltd. He has over 200 publications on metallography, failure analysis, quantitative metallography, and image analysis. He has taught at several universities and for the ASM MEI program. This course provides an overview of factors that produce microstructure and control its formation, amount, size and morphology for both nonferrous and ferrous alloys. The basic premise of physical metallurgy is that there is a relationship among composition and processing that creates microstructure, and that microstructure has a strong influence on many properties and service behavior. The course will illustrate the evolution of microstructures from solidification, through processing and heat treatment, and will highlight ways to properly reveal the microstructure. |