Grant Abstracts 2010   Arizona State University Deirdre Meldrum, Roger Johnson, Laimonas Kelbauskas, Lea Sistonen Tempe, AZ $1,000,000 2010 This four-year project aims to build a novel, live-cell imaging instrument for basic and clinical science application. The 3D microscope called the “cell CT scanner” will provide functional images, revealing the molecular mechanisms underlying important metabolic and disease processes. Cell CT is analogous to diagnostic radiology CT in that it will reconstruct 3D images with isotropic, submicron resolution from hundreds of projections acquired from many angles as the cell is rotated. A key component of the research is to determine the best method for rotating cells, which must be done extremely precisely without harming the cell. The team will investigate several cell rotation methods including one that rotates an optically trapped cell in a microfluidic vortex and another that uses an infrared light beam having asymmetrical intensity distribution. They will use fluorescent antibody probes and fusion-protein constructs specific to the proteins of interest to label cells for emission CT scanning. The technology will be validated by studying cells from immortalized cell lines representative of various stages of epithelial cancers, and cells disaggregated from human biopsies spanning the same disease spectrum. Cell CT may enable for the first time rapid spatial localization of proteins, and assessment of their concentrations in subcellular compartments and microdomains, providing insights concerning relationships between cell structure, function and disease.   Boston College Michael Naughton, Kris Kempa Chestnut Hill, MA $1,000,000 2010 The project is to develop a unique and powerful approach to nanoscale optical imaging, bridging near-field and far-field optics with deep sub-diffraction-limit resolution. When developed, this nanoscale coaxial optical microscope (NCOM) technique will facilitate rapid nanoscale optical imaging of micro- to macroscopic landscapes, under optical intensities well below those requiring laser illuminations, including the possibility of label-free imaging. The basic concept is a novel superlensing, guided wave metamaterial, consisting of an array of nanoscale coaxial waveguides. This array is arranged such that the spacing between the wire ends on one lens surface is smaller than the wavelength λ of visible light, while that on the other side exceeds λ. Each “nanocoax” at the object side collects light in its vicinity via an optical antenna effect and propagates it along the waveguide to the image side, where it emerges into the optical far-field. A conventional optical microscope, focused onto this projected image, can now resolve features with significantly greater resolution and lower optical intensity than current state-of-the-art techniques. This transformative NCOM technique, capable of noninvasively resolving features smaller than the diffraction limit, will find enormous utility in biological / living systems, in every field in medicine, including genetics, genomics, oncology, and pharmacology, and with wide applicability in the physical sciences.   University of California, Los Angeles David Eisenberg, James Bowie, Duilio Cascio, Mari Gingery, Michael Sawaya, Todd Yeates Los Angeles, CA $900,000 2010 This project will develop tools having the potential to better understand cells in health and disease by providing precise pictures of the interacting molecules. Nano X-ray diffraction can reveal the 3D atomic structure of intracellular organelles and aggregates that mediate the metabolism and pathology of cells. In contrast, nearly all present information on the atomic structure of cellular constituents has come from purified molecules, removed from cells. The research group will exploit recent advances in the production of highly focused (nano) X-ray beams and free electron lasers, directing these beams onto biological cells and subcellular organelles, prepared by new methods for X-ray examination, and devise methods for collecting and interpreting the diffraction data. One application will be to learn the atomic structure of the carboxysome, the organelle that removes CO2 from the atmosphere, a process that is critical to sustaining life on earth as we know it. A second application will be to learn which types of cells contain aggregates of protein in the amyloid state, a state similar to the deposits formed in Alzheimer’s and Parkinson’s diseases, but in some cases apparently part of normal function. The same tools will be applied to microcrystals and microdomains of larger crystals with the potential to gain significantly more information than is presently possible.   University of California, Santa Barbara Sumita Pennathur, Paul Atzberger, Andrew Cleland, Frederic Gibou, Todd Squires Santa Barbara, CA $1,000,000 2010 A technology able to separate, detect and analyze nanoparticles and molecules according to size and charge has significant implications for future medical diagnostics and nanoparticle technology. If these operations could be performed cheaply and quickly, the technology could result in new tools to be used, for instance, in a doctor’s office for rapid and precise molecular based diagnoses. At the same time, applications toward efficient nanoparticle and molecular synthesis, purification, and characterization could be of major importance to the burgeoning nanomanufacturing industry. The goal is to develop a new platform capable of detection, analysis and sorting of biological and synthetic nanoparticles, based on the integration of two complementary techniques: 1) Size-based detection and sorting of individual unlabeled nanoparticles, and 2) gel-free electrophoretic separation of nanoparticles in nanochannels. The interdisciplinary nature of the group has a diversity of perspectives offering the potential for new insights and approaches for the development of novel solutions.   University of Texas, Austin Rodney Ruoff Austin, TX $1,000,000 2010 The goal is to enable large-scale fabrication of graphene/ultrathin graphite (‘G/UG’) films/foils. The influence of structure and particularly grain boundaries on the physical properties of such foils will be elucidated. Studies on processing (delaminating, cutting, folding, laminating with other films, making fiber strips, wetting, etc.) will accelerate the use of these new materials in existing applications and foster innovation in new applications. A new method of heating the metal substrates that G/UG films grow on will be implemented and optimized to produce material with minimum defects, largest grains and best grain boundaries. Optimized G/UG foils are not expected to be brittle and their specific strengths may far exceed those of any materials now available. This will motivate their use in new structural applications, as well as in thermal management, as compliant transparent conductive electrodes, in nanoelectronics (the carrier mobility in graphene is very high), in micro- and nano-electromechanical systems as actuators, strain gauges, pressure sensors and others. Applications will be realized through active collaborations and an ensuing effort will help to ensure benefit to society through the eventual factory-level production of G/UG materials.   Yale University Alanna Schepartz, Scott Miller New Haven, CT $1,000,000 2010 Yale researchers propose an innovative approach to achieve a long-standing goal in the field of chemistry – the construction of bona fide artificial enzymes.  It will apply methods from four related fields: organic chemistry, carbohydrate chemistry, enzymology, and biophysics.  Catalytic substrates known as β-peptides bundles will be created to mimic carbohydrate-processing enzymes.  The investigators will optimize three steps that define the catalytic cycle: (1) substrate binding, (2) chemical reactivity, and (3) product release.  To optimize substrate binding, β peptide bundles will be assembled that contain an extended phenyl boronic acid “docking site.”  To optimize chemical reactivity, the researchers will assemble in parallel β-peptide bundles appended with 2-3 carboxylate side chains (of various structure and acidity) in positions that mimic their spacing in carbohydrate processing enzymes.  Optimizing the product release will make use of traditional or stopped-flow kinetics methods as well as surface plasmon resonance.  These features will then be combined into first-generation catalysts to optimize their properties through the application of combinatorial, catalytic, activity-based screens.  These catalysts will then be optimized through the application of combinatorial, catalytic activity-based screens and studied in detail using both biophysical (calorimetry, crystallography) and spectroscopic (NMR) methods to identify and explore relationships between catalytic efficiency, cooperative folding, and protein dynamics.