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.
|Kong, Linghao; Murata, Michael M; Digman, Michelle A (2018) Absence of REV3L promotes p53-regulated cancer cell metabolism in cisplatin-treated lung carcinoma cells. Biochem Biophys Res Commun 496:199-204|
|Mäntylä, Elina; Chacko, Jenu V; Aho, Vesa et al. (2018) Viral highway to nucleus exposed by image correlation analyses. Sci Rep 8:1152|
|Davey, Rhonda J; Digman, Michelle A; Gratton, Enrico et al. (2018) Quantitative image mean squared displacement (iMSD) analysis of the dynamics of profilin 1 at the membrane of live cells. Methods 140-141:119-125|
|Malacrida, Leonel; Gratton, Enrico (2018) LAURDAN fluorescence and phasor plots reveal the effects of a H2O2 bolus in NIH-3T3 fibroblast membranes dynamics and hydration. Free Radic Biol Med 128:144-156|
|Malacrida, Leonel; Rao, Estella; Gratton, Enrico (2018) Comparison between iMSD and 2D-pCF analysis for molecular motion studies on in vivo cells: The case of the epidermal growth factor receptor. Methods 140-141:74-84|
|Hedde, Per Niklas; Gratton, Enrico (2018) Selective plane illumination microscopy with a light sheet of uniform thickness formed by an electrically tunable lens. Microsc Res Tech 81:924-928|
|Kobylkevich, Brian M; Sarkar, Anyesha; Carlberg, Brady R et al. (2018) Reversing the direction of galvanotaxis with controlled increases in boundary layer viscosity. Phys Biol 15:036005|
|Sameni, Sara; Malacrida, Leonel; Tan, Zhiqun et al. (2018) Alteration in Fluidity of Cell Plasma Membrane in Huntington Disease Revealed by Spectral Phasor Analysis. Sci Rep 8:734|
|Mah, Emma J; Lefebvre, Austin E Y T; McGahey, Gabrielle E et al. (2018) Collagen density modulates triple-negative breast cancer cell metabolism through adhesion-mediated contractility. Sci Rep 8:17094|
|Ranjit, Suman; Malacrida, Leonel; Gratton, Enrico (2018) Differences between FLIM phasor analyses for data collected with the Becker and Hickl SPC830 card and with the FLIMbox card. Microsc Res Tech 81:980-989|
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