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Organizer: Maryann Martone |
Speakers: Paul Wiseman |
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This tutorial will be an introduction to fl uorescence correlation spectroscopy (FCS) with a particular focus on its imaging analog called image correlation spectroscopy (ICS). An overview of the history and theory of these fl uctuation microscopy methods will be presented along with a description of the microscopy instrumentation required to perform such measurements. The second half of the tutorial will describe biological applications of image correlation techniques in both cell biology and neuroscience. We will discuss specifi c applications such as measuring protein transport (both fl ow and diffusion), receptor clustering and interactions for proteins involved in the adhesion machinery of adherent and migratory cells. The tutorial will also discuss applications in neuroscience for measurement of dendritic spine densities in hippocampus tissue slices. |
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Speaker:Guido Gaietta |
| Since their fi rst introduction in the early '80s, fl uorescent proteins have proven incredibly valuable to the study of many proteins directly in their native environment. Recent advancements in molecular tagging have given us the ability to follow a target protein at different resolution levels, to examine its interactions with other proteins and ultimately, to study its function. This tutorial will provide a general overview of two tagging systems: GFP (and its variants) and 4Cys/FlAsH. We will examine the basics of each system, compare them with one another and explore their applications to confocal, 2-photon and electron microscopy. Examples of FRET, FRAP and FALI will also be covered during the presentation. |
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Speaker:Daniel A. Peterson |
| Quantitative histology has become an increasingly important form of analysis in biomedical research. Some form of outcome measurement, frequently cell number, is required to provide data for statistical analysis to accept or reject the hypothesis. For microscopic analysis, this requires counting objects (cells) that are embedded within a larger structure (tissue). However, these cells must be counted from sections produced from the tissue. As a result of sectioning the tissue and visualizing the cells through an objective lens that produces a depth of fi eld, one must consider potential artifacts that could introduce error into the determination of the number of objects. These sources of error can be addressed through the use of systematic three-dimensional sampling probes collectively known as design-based stereology. This tutorial will review the principles involved and illustrate practical examples of implementing design-based stereology. |
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Organizer: Peter Crozier |
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Speaker: Dale E. Newbury |
| Energy dispersive x-ray spectrometry (EDS) performed in the scanning electron microscope (SEM) has become a basic procedure supporting a vast range of applications. Most often applied to practical problems as a qualitative analysis technique, SEM/ EDS is actually capable of achieving quantitative results with a small error budget, +5% relative or better. The steps to achieve this level of accuracy with SEM/EDS will be discussed: (1) careful measurement science practices for electron-excited x-ray spectrometry; (2) robust qualitative analysis at all levels: major (C > 0.1 mass fraction), minor (0.01 < C < 0.1), and trace (C < 0.01); (3) statisticallystable peak and background deconvolution; and (4) quantitative matrix corrections. |
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Speaker: Jian-Min Zuo |
| Electron diffraction patterns are routinely recorded in electron microscopes to obtain quantitative structure information about crystal phases and orientation relationships. The recent developments of fi eld-emission gun electron microscopes, energy-fi lters, digital detectors and computer simulations have signifi cantly improved both the sensitivity and quantifi cation of electron diffraction. Meanwhile, new materials development focuses increasingly on new structures at nanoscale, where the spatial resolution of electron diffraction is an advantage. This tutorial starts with basic knowledge of electron diffraction and then covers its application for modern materials characterization. Topics include: diffraction modes in TEM, crystal diffraction and indexing, the convergent beam geometry and diffraction of nanomaterials. The level of the tutorial is aimed at TEM users who have practical knowledge of electron diffraction, but are interested in improving their skills on interpretation and learning new quantitative techniques of electron diffraction. |
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Speaker: David. A. Muller |
| Modern commercial field emission transmission electron microscopes (TEM) have the potential to produce atomic-resolution scanning transmission electron microscopy (STEM) images if properly aligned and installed. This tutorial will cover practical and theoretical aspects necessary to optimize the microscope for STEM. Topics covered: interpreting Ronchigrams, choosing the right apertures and camera lengths, bright and dark fi eld imaging, how to make your STEM images look like they were taken on a TEM, Z-contrast and strain mapping, sample thickness dependence of STEM images and imaging artifacts. |
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Speaker: Molly McCartney |
| Electron holography is an evolving technique that is gaining increased interest in the microscopy community for characterization of electric fi elds and magnetic materials in addition to its original application for resolution enhancement. This tutorial will cover a wide range of aspects including the history of the technique and theoretical and experimental issues. In particular, the source and implications of the phase shifts measured in the reconstructed holograms will be discussed. Emphasis will be placed on practical aspects of electron holography including the experimental set-up, reconstruction and the precision and accuracy of the reconstructed phase images. Applications to semiconductor junction delineation, piezoelectric fi elds and magnetic thin fi lms will be discussed. |
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Organizers: Peter Crozier and Maryann Martone |
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Speaker: Mike Marko |
| There are a wide variety of applications for electron tomography, and a correspondingly wide variety of imaging modes and data collection strategies. In biological sciences, low-dose, low-temperature tomography of unstained frozen-hydrated specimens is of great interest, while tomography of stained, thick plastic sections at high accelerating voltage is essential for other studies. In materials science, techniques such as HAADF STEM tomography are needed for crystalline specimens. Elemental mapping using EFTEM or STEM is being applied in both fi elds. The tutorial will attempt to cover all known forms of transmission electron tomography, and describe how each is implemented for an appropriate application. |
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Organizer: Lucille A. Giannuzzi |
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The Ask-the-Experts session takes on a different format this year to provide programming closer to the time of the annual Microscopy and Microanalysis (M&M) meeting, and to better address the needs of the meeting attendees. Rather than listing topics in the Call for Papers, which must be produced nearly a year before the meeting, an opportunity to submit questions to experts attending M&M 2004 will be provided at the time the program is posted on the meeting web site in the months leading up to the meeting. A link will allow participants to submit questions, and name the expert to whom the question is addressed, as they browse the M&M 2004 program on-line. The Ask-the- Experts organizers will then seek to schedule a time to address topics of signifi cant interest during the meeting. The schedule of topics will be announced and posted to the meeting website a couple of weeks before the early registration deadline, so that potential M&M 2004 attendees can schedule in the topics of interest to them. Please plan to participate both in the topic selection and the sessions, so that we can gauge whether this change of format is beneficial! |