This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project seeks to characterise the real time diffusion dynamics of fluoresently labelled actin molecules within the nuclei of living cultured mammalian cells.
We aim to determine if nuclear Actin molecules exhibit evidence of self binding (polymerization) and if they show binding or barriers to diffusion in relation to nuclear components (chromatin and transcription factors). The major experiments for this work include; 1) cell culture of L6 Myoblast adherent cells using standard culture reagents and techniques. 2) Transfection of the cultured cells with fluorescently labelled actin plasmid DNA. 3) Confocal imaging and spectroscopy of cells expressing fluorescently labelled actin molecules. Spectroscopy tecniques used will include scanning fluorescence correlation spectroscopy, rast image correlation spectroscopy, number and brightness analysis, and photon counting histogram.
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|Digiacomo, Luca; Digman, Michelle A; Gratton, Enrico et al. (2016) Development of an image Mean Square Displacement (iMSD)-based method as a novel approach to study the intracellular trafficking of nanoparticles. Acta Biomater 42:189-198|
|Malacrida, Leonel; Astrada, Soledad; Briva, Arturo et al. (2016) Spectral phasor analysis of LAURDAN fluorescence in live A549 lung cells to study the hydration and time evolution of intracellular lamellar body-like structures. Biochim Biophys Acta 1858:2625-2635|
|Chen, Hongtao; Gratton, Enrico; Digman, Michelle A (2016) Self-assisted optothermal trapping of gold nanorods under two-photon excitation. Methods Appl Fluoresc 4:035003|
|Golfetto, Ottavia; Hinde, Elizabeth; Gratton, Enrico (2015) The Laurdan spectral phasor method to explore membrane micro-heterogeneity and lipid domains in live cells. Methods Mol Biol 1232:273-90|
|Willenberg, Rafer; Steward, Oswald (2015) Nonspecific labeling limits the utility of Cre-Lox bred CST-YFP mice for studies of corticospinal tract regeneration. J Comp Neurol 523:2665-82|
|Chen, Hongtao; Gratton, Enrico; Digman, Michelle A (2015) Spectral properties and dynamics of gold nanorods revealed by EMCCD-based spectral phasor method. Microsc Res Tech 78:283-93|
|Scarlata, Suzanne; Golebiewska, Urszula (2014) Linking alpha-synuclein properties with oxidation: a hypothesis on a mechanism underling cellular aggregation. J Bioenerg Biomembr 46:93-8|
|Pang, Yan Ling Joy; Poruri, Kiranmai; Martinis, Susan A (2014) tRNA synthetase: tRNA aminoacylation and beyond. Wiley Interdiscip Rev RNA 5:461-80|
|Sharma, Himanshu; Digman, Michelle A; Felsinger, Natasha et al. (2014) Enhanced emission of fluorophores on shrink-induced wrinkled composite structures. Opt Mater Express 4:753-763|
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