This proposal seeks to continue and expand technological biomedical research, collaborative, service, training and dissemination activities ofthe Laboratory for Fluorescence Dynamics (LFD). Since 1986, the LFD has been a national resource center dedicated to 1) develop of new fluorescence technologies for biology and medicine and 2) service, training, and dissemination of fluorescence methods in a user facility. In previous years (1986-2010), we established our reputation as the leaders in technological development for fluorescence dynamics. Since moving to UCI in 2006, we expanded the core research and collaborative work in this new fertile biomedical research environment. We created stmctures for training through workshops, tutorials and organization of LFD training courses at UCI and other US universities. Core development expands current limits of biomedically relevant fluorescence instrumentation, particularly for the study of cellular processes. Projects include: 1) new concepts in fluorescence microscopy that develop novel imaging methods to exploit the dynamic fluorescence methodologies that are the hallmark ofthe LFD developments, 2) expanding the concept of spatio-temporal fluctuation correlation analysis to a new powerful image approach that provides detailed information of molecular flow at the nanoscale, 3) new methods for 3D nanoimaging based on the orbital tracking approach, 4) novel technologies for fluorescence lifetime imaging analysis that allow new contrast mechanisms to be applied to tissues and animals, and 5) development of software/algorithm platform for disseminating the methods for data collection and analysis developed at the LFD. Users atthe LFD find ready access to new and innovative technology, which affords them a unique opportunity to rapidly advance their own research programs. Driving biological problems exploit and push technological advances and research in biological processes, macromolecular assembly, tissue organization and dynamics, membrane morphology/function relationships, and innovative biomedical instrumentation. Educational programs include hands-on training of students, postdoctoral fellows and visiting scientists, workshops and specialized schools in innovative fluorescence methodologies.
The proposed technological developments provide real time detection and localization of molecular interactions in 3D in live cells and tissues. We have obtained technological breakthroughs in the areas of microscopy imaging, algorithm developments for revealing comprehensive spatio-temporal correlations, nanoimaging methods with unprecedented resolution and new spectroscopic contrast methods that can identify metabolic states of cells and predict their behavior.
|Hedde, Per Niklas; Stakic, Milka; Gratton, Enrico (2014) Rapid measurement of molecular transport and interaction inside living cells using single plane illumination. Sci Rep 4:7048|
|Crosignani, Viera; Jahid, Sohail; Dvornikov, Alexander S et al. (2014) A deep tissue fluorescence imaging system with enhanced SHG detection capabilities. Microsc Res Tech 77:368-73|
|James, Nicholas G; Digman, Michelle A; Ross, Justin A et al. (2014) A mutation associated with centronuclear myopathy enhances the size and stability of dynamin 2 complexes in cells. Biochim Biophys Acta 1840:315-21|
|Ranjit, Suman; Dvornikov, Alexander; Holland, David A et al. (2014) Application of three-photon excitation FCS to the study of protein oligomerization. J Phys Chem B 118:14627-31|
|Bachir, Alexia I; Zareno, Jessica; Moissoglu, Konstadinos et al. (2014) Integrin-associated complexes form hierarchically with variable stoichiometry in nascent adhesions. Curr Biol 24:1845-53|
|Kang, Dong-Ku; Ali, M Monsur; Zhang, Kaixiang et al. (2014) Rapid detection of single bacteria in unprocessed blood using Integrated Comprehensive Droplet Digital Detection. Nat Commun 5:5427|
|Pate, Kira T; Stringari, Chiara; Sprowl-Tanio, Stephanie et al. (2014) Wnt signaling directs a metabolic program of glycolysis and angiogenesis in colon cancer. EMBO J 33:1454-73|
|Annibale, Paolo; Gratton, Enrico (2014) Advanced fluorescence microscopy methods for the real-time study of transcription and chromatin dynamics. Transcription 5:|
|Bonaventura, Gabriele; Barcellona, Maria Luisa; Golfetto, Ottavia et al. (2014) Laurdan monitors different lipids content in eukaryotic membrane during embryonic neural development. Cell Biochem Biophys 70:785-94|
|Plotegher, N; Gratton, E; Bubacco, L (2014) Number and Brightness analysis of alpha-synuclein oligomerization and the associated mitochondrial morphology alterations in live cells. Biochim Biophys Acta 1840:2014-24|
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