Over the last two years, fluorescence microscopy has experienced an order of magnitude improvement in resolution via single molecule based imaging techniques [296]. Evolution in the technologies of detectors, computational power, molecular biology and fluorescence probes now allows the brute force approach of localizing thousands of individual probes that are labeling specific targets in vivo. The basic concept of this imaging technique is that single molecules can be localized with an accuracy that scales roughly as sigma(loc)=sigma psf/N[1/2], where sigma (ioc) is the localization accuracy sigma psf is the width of the microscope point spread function, and N is the expected number of collected photons from the single molecule [297]. Recent developments in bright photo-activatable, photo-switchable and naturally intermittent fluorescent probes have allowed high densities of single fluorescent molecules to be imaged and localized individually with accuracy approaching 10 nm, leading to super-resolution images, in fixed or neariy static structures. This concept is illustrated in Figure 56. SML-SR has been demonstrated by several groups [26,298,299] with variations based on the probes used, imaging conditions and analysis approaches. Single Molecule Localization based Super-Resolution (SML-SR) techniques join SPT and single molecule imaging as the most powerful available tools to access protein-protein dynamics at the 10 nm scale in live and fixed cells. One important advantage of SML-SR over other SR approaches (such as STED [300], and Saturated Patterned Excitation [301] is that positions of labeled targets (used to generate the SR images) can be used directly for analysis of clustering and co-clustering as done with immuno-gold labeling in EM [11]. The primary goals of the SR-core are to (1) provide these state of the art single molecule fluorescence techniques to the STMC and (2) develop new techniques that allow access to ever smaller spatial and temporal scales. The SR-core personnel will work closely with the biologists and the analysis core to design and execute single molecule experiments to directly answer questions described elsewhere in this proposal.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center (P50)
Project #
5P50GM085273-05
Application #
8534178
Study Section
Special Emphasis Panel (ZGM1-CBCB-4)
Project Start
Project End
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
5
Fiscal Year
2013
Total Cost
$165,199
Indirect Cost
$55,066
Name
University of New Mexico Health Sciences Center
Department
Type
DUNS #
829868723
City
Albuquerque
State
NM
Country
United States
Zip Code
87131
Hoard, Brittany; Jacobson, Bruna; Manavi, Kasra et al. (2016) Extending rule-based methods to model molecular geometry and 3D model resolution. BMC Syst Biol 10 Suppl 2:48
Kerketta, Romica; Halász, Ádám M; Steinkamp, Mara P et al. (2016) Effect of Spatial Inhomogeneities on the Membrane Surface on Receptor Dimerization and Signal Initiation. Front Cell Dev Biol 4:81
Levin, Drew; Forrest, Stephanie; Banerjee, Soumya et al. (2016) A spatial model of the efficiency of T cell search in the influenza-infected lung. J Theor Biol 398:52-63
Winner, Kimberly R Kanigel; Steinkamp, Mara P; Lee, Rebecca J et al. (2016) Spatial Modeling of Drug Delivery Routes for Treatment of Disseminated Ovarian Cancer. Cancer Res 76:1320-34
Meddens, Marjolein B M; Liu, Sheng; Finnegan, Patrick S et al. (2016) Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution. Biomed Opt Express 7:2219-36
Hansen, Scott G; Wu, Helen L; Burwitz, Benjamin J et al. (2016) Broadly targeted CD8⁺ T cell responses restricted by major histocompatibility complex E. Science 351:714-20
Marjon, K D; Termini, C M; Karlen, K L et al. (2016) Tetraspanin CD82 regulates bone marrow homing of acute myeloid leukemia by modulating the molecular organization of N-cadherin. Oncogene 35:4132-40
Fricke, G Matthew; Letendre, Kenneth A; Moses, Melanie E et al. (2016) Persistence and Adaptation in Immunity: T Cells Balance the Extent and Thoroughness of Search. PLoS Comput Biol 12:e1004818
Lin, Jia; Wester, Michael J; Graus, Matthew S et al. (2016) Nanoscopic cell-wall architecture of an immunogenic ligand in Candida albicans during antifungal drug treatment. Mol Biol Cell 27:1002-14
Termini, Christina M; Lidke, Keith A; Gillette, Jennifer M (2016) Tetraspanin CD82 Regulates the Spatiotemporal Dynamics of PKCα in Acute Myeloid Leukemia. Sci Rep 6:29859

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