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.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Specialized Center (P50)
Project #
Application #
Study Section
Special Emphasis Panel (ZGM1-CBCB-4)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of New Mexico Health Sciences Center
United States
Zip Code
Chylek, Lily A; Akimov, Vyacheslav; Dengjel, Jörn et al. (2014) Phosphorylation site dynamics of early T-cell receptor signaling. PLoS One 9:e104240
Chylek, Lily A; Harris, Leonard A; Tung, Chang-Shung et al. (2014) Rule-based modeling: a computational approach for studying biomolecular site dynamics in cell signaling systems. Wiley Interdiscip Rev Syst Biol Med 6:13-36
Wu, Meiye; Piccini, Matthew E; Singh, Anup K (2014) MiRNA detection at single-cell resolution using microfluidic LNA flow-FISH. Methods Mol Biol 1211:245-60
Itano, Michelle S; Graus, Matthew S; Pehlke, Carolyn et al. (2014) Super-resolution imaging of C-type lectin spatial rearrangement within the dendritic cell plasma membrane at fungal microbe contact sites. Front Phys 2:
Lund, Troy C; Patrinostro, Xiaobai; Kramer, Ashley C et al. (2014) sdf1 Expression reveals a source of perivascular-derived mesenchymal stem cells in zebrafish. Stem Cells 32:2767-79
Liu, Sheng; Lidke, Keith A (2014) A multiemitter localization comparison of 3D superresolution imaging modalities. Chemphyschem 15:696-704
Steinkamp, Mara P; Low-Nam, Shalini T; Yang, Shujie et al. (2014) erbB3 is an active tyrosine kinase capable of homo- and heterointeractions. Mol Cell Biol 34:965-77
Westrate, Laura M; Drocco, Jeffrey A; Martin, Katie R et al. (2014) Mitochondrial morphological features are associated with fission and fusion events. PLoS One 9:e95265
Valley, Christopher C; Lidke, Keith A; Lidke, Diane S (2014) The spatiotemporal organization of ErbB receptors: insights from microscopy. Cold Spring Harb Perspect Biol 6:
Jerman, Stephanie; Ward, Heather H; Lee, Rebecca et al. (2014) OFD1 and flotillins are integral components of a ciliary signaling protein complex organized by polycystins in renal epithelia and odontoblasts. PLoS One 9:e106330

Showing the most recent 10 out of 50 publications