Continuing support is proposed for the Laboratory for Fluorescence Dynamics to expand technological biomedical research to new fields of discovery and applications and to strengthen collaborative, service, training and dissemination activities for the benefit of the biomedical research community. Fluorescence imaging and fluorescence methodologies have reached a prominent role in Biology, Biophysics and in Clinical Research. The LFD is a biotechnological resource center with a history of successes in developing new fluorescence technologies for biology and medicine and with a strong commitment to service, training, and dissemination components aimed at bringing fluorescence methods in a user facility available to the biomedical research community. For this proposal we have conceived a strong technical development program to expand current limits of imaging fluorescence instrumentation, particularly for the study of dynamical cellular processes. Projects include: developments of new microscope platforms for imaging in deep tissues, still maintaining diffraction limited resolution; exploiting the light sheet microscope for fluctuation analysis with the potential for developing radical new approaches to measure molecular flow in cells and tissue at the nanoscale; using new electronics and optics for 3D imaging of cellular dynamics; expanding the phasor approach to fluorescence lifetime imaging analysis to spectral and diffusion analysis for fast, fit-free visualization of dynamical processes and expanding our software to allow new algorithms for data acquisition and analysis in our new microscopes. The organization structure of the LFD allows users and collaborators to access innovative technologies only available in this BTRR resource, which affords them a unique opportunity to rapidly advance their own research programs. A rich program of 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 for detection of bacteria in blood which could have an impact in the clinical area. Educational programs proposed for this application include hands-on training of students, postdoctoral fellows and visiting scientists, workshops and specialized schools in innovative fluorescence methodologies. Our improved web site will continue to be a reference source for information about fluorescence dynamic techniques, and a large repository of publications, lectures and tutorials.

Public Health Relevance

The proposed technological developments are relevant to a growing number of problems in biomedical research that require real time detection and localization of molecular interactions in 3D in live cells and tissues at the single molecule level. The visualization analysis program developed in this application allows single cells analyses of metabolic indices of cells in the context of tissues, a significant advance in tissue analysis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Biotechnology Resource Grants (P41)
Project #
5P41GM103540-32
Application #
9320689
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
Budget Start
2017-07-01
Budget End
2018-06-30
Support Year
32
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of California Irvine
Department
Type
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92617
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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

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