2013 Eastern Analytical Symposium & Exposition Analytical in Motion Knowledge › Network › Career Abstract Book November 18–20, 2013 Garden State Exhibit Center Somerset, New Jersey www.EAS.org 13974-EAS_8.5x11_2013Cover-AbstractBook_PREP.indd 1 10/15/13 5:00 PM 2013 EAS Abstracts November 2013 2013 EAS Abstracts This volume contains the final abstracts for the oral and poster presentations which take place on Monday, November 18, through Wednesday, November 20, 2013. For each abstract provided, a complete mailing address for the presenting author is shown. Additional authors are indicated, however, their mailing addresses are not provided. Schedule and meeting room information for the technical sessions, as well as information concerning short courses, exhibitor workshops, and the exposition, are contained in the Final Program Book. More Information To obtain answers to EAS-related questions after the meeting: EAS Hotline 732-449-2280 EAS E-mail [email protected] EAS Web Site www.EAS.org Eastern Analytical Symposium & Exposition, Inc. P.O. Box 185 Spring Lake, NJ 07762 Save the Date The 2014 EAS November 17 - 19, 2014 Somerset, NJ We want you to be a part of the 53rd Eastern Analytical Symposium! 2014 Call for Papers March 1 – April 15, 2014 1 2013 EAS Abstracts November 2013 ABSTRACTS OF TECHNICAL PAPERS 1 Synchrotron X-Ray Spectroscopy - How 10 Orders of Magnitude 4 The Good, the Bad and the Ugly: Understanding the Roles of Makes Hard Things Easy Metals in Biology Using Synchrotron Radiation Stephen P. Cramer, University of California, Lawrence Berkeley Lab, Graham N. George, University of Saskatchewan, 114 Science Pl., Department of Chemistry & Physical Biosciences Division, Davis, CA Sasaktoon, SK S7N5E2, Canada 95616 One of the major strengths of X-ray absorption spectroscopy (XAS) is its ability to Over the past 40 years, the brightness of synchrotron radiation sources has in- probe chemical form in any sample without any chemical pre-treatment. The low creased by about 10 orders of magnitude. It is still improving today. In this talk I give concentrations of metals and metalloids in biological tissues have meant that in examples of experiments that were once heroic and that are now almost routine. the past XAS has been difficult to apply to these samples. Recent developments in I start with the EXAFS of MoFe nitrogenase, comparing early data with the latest synchrotron beam line and detector technology mean that XAS of intact biological analysis and the 1 Å crystal structure. By combining the EXAFS Debye-Waller factor samples is now achievable. XAS can reveal the molecular details of metals within with empirically calibrated molecular force-fields, we can obtain average distances living systems, how they interact with one another, how they confer beneficial prop- and rms disorder both with ~0.01 Å accuracy. I then discuss progress in soft X- erties, and how they act as poisons. As most tissues have spatial structure, there is ray spectroscopy and detectors, showing the potential going forward for arrays of great additional interest in visualizing the spatial location of the metal or metalloid hundreds of superconducting tunnel junctions to probe highly dilute systems. Next, as well as its chemical forms. The use of microfocus X-ray beams gives the oppor- I compare our earliest efforts at “high-resolution” X-ray fluorescence at 1 eV resolu- tunity of producing maps of elements and of specific chemical types of elements in tion with recent work on inelastic X-ray scattering a 1 meV resolution. I conclude dilute biological systems. These developments are discussed, drawing upon recent with recent progress in nuclear inelastic scattering. In 1995, Seto did the first ex- examples from our research on XAS of toxic elements in biological tissues. periment at 7 meV resolution on pure Fe metal. We are now looking at modes in metalloproteins at sub-millivolt resolution from samples with millimolar Fe. I end with 5 Analysis of Vesicle Contents with Electrochemistry and Mass some prognostication of what might be possible in the future in the current bright- Spectrometry ness trend continues. Andrew G. Ewing, Chalmers University and Gothenburg University, Kemivagen 10, Gothenburg 41296, Sweden 2 X-Ray Emission Spectroscopy - A Powerful Tool to Study Amperometry can be used to measure individual exocytosis events from secreting Biocatalysts cells. We have analyzed amperometric peaks corresponding to release at PC12 Uwe Bergmann, SLAC National Accelerator Laboratory, 2575 Sand Hill cells and found stable plateau currents during the decay of the peaks, indicating Rd., Menlo Park, CA 94025 closing of the vesicle after incomplete release of the vesicular content. From careful Much of the chemistry essential to life is governed by catalytic reactions that take analysis of these data, we have now proposed a process for most exocytosis events place at metal centers within biological systems. Among the various tools to study where the vesicle partially opens to release transmitter and then closes directly such metal centers X-rays are particularly powerful due to their sensitivity to the again, leaving the possibility for both a stable pre- and post-spike foot to be ob- geometric and electronic structure at an atomic level. X-ray emission spectroscopy served with amperometry. The experiments correlate well with those from the elec- (XES) is such a probe and with the advent of very powerful X-ray sources and novel trochemical cytometry method we have developed to count electroactive molecules X-ray spectrometers the technique has now been successfully applied to study the in individual synaptic vesicles in directly sampled populations from cells or brain extremely weak X-ray spectral signals from some of the most important biocata- tissue. With this method we can compare the total numbers of molecules to those lysts. We introduce the technique, present the state-of-the-art X-ray sources and released and only a fraction is in fact released both at cell models and in mammalian instrumentation, and discuss research examples. In particular we show how XES brains. We have combined these experiments with mass spectrometry imaging to has provided a breakthrough in the understanding of the atomic structure of the compare the concentration of transmitters across single vesicles. iron-molybdenum co-factor in nitrogenase, the enzyme used by some organisms to fix atmospheric nitrogen gas by breaking the N≡N triple bond, one of the strongest 6 Autophagy from an Individual Organelle Perspective bonds in nature. The other example focuses on a series of studies of the MnCaO Edgar A. Arriaga, University of Minnesota, 207 Pleasant St. SE, cluster in photosystem II, responsible for splitting water into molecular oxygen. Be- Minneapolis, MN 55455, Chad P. Satori sides using synchrotron X-ray sources for many years, we have recently started Autophagy is responsible for the degradation of damaged intracellular components. to perform studies at the Linac Coherent Light Source, the world’s first X-ray free Current measurements, such as those done by Western blotting, are insufficient to electron laser. We discuss our first results and show how this revolutionary machine understand autophagy and its roles in aging and disease. Capillary electrophoresis can be used to study the photosynthetic water splitting mechanism in real-time. coupled to laser induced fluorescence detection (CE-LIF) has been used previously to count and determine properties of individual organelles, but has never been used 3 Coherent and Inelastic Resonant X-Ray Scattering on autophagy organelles or for determination of temporal changes of such proper- Esen E. Alp, Argonne National Laboratory, Advanced Photon Source, ties. In this presentation we report on the use of this technique to investigate au- Bldg. 431-D004, Argonne, IL 60439 tophagy in L6 cells, a skeletal muscle model. Autophagy organelles were isolated Five decades ago, Nobel Prize was given to R. Mössbauer for the discovery of from L6 cells expressing GFP-LC3 and analyzed by CE-LIF for the determination of the eponymous effect. For the last three decades, many groups around the world the number of autophagy organelles, their individual GFP-LC3 fluorescence intensi- started research programs to make use of synchrotron radiation to replace the ra- ties, and their individual electrophoretic mobilities. Basal and rapamycin-driven au- dioactive parent isotope as a source. The early efforts were amply rewarded with the tophagy were compared under steady state and dynamic conditions. Under steady arrival of high-brightness third generation synchrotron radiation sources. Nuclear state conditions, there were differences in organelle number and electrophoretic resonant X-ray scattering of synchrotron radiation is now a useful tool for investigat- mobility distributions. Under dynamic conditions, the temporal trends of individual ing thermodynamic properties of organic and inorganic materials in various scien- organelle properties upon treatment with vinblastine were the opposite of the trends tific disciplines including condensed matter physics, materials science, biophysics, observed under basal autophagy. Individual organelle analysis by CE-LIF is a pow- mineral physics and bioinorganic chemistry and geochemistry. Interesting studies erful technology to investigate the complexity and temporal nature of autophagy, a of iron proteins and enzymes, and their model compounds like single crystal por- process that plays critical roles in response