Fluorescence Spectroscopy is inherently a very sensitive technique; it already forms the basis of most non-radioactive real time assays like PCR. Our lab previously collaborated with former fellows (now in biotech industry) to develop alternatives to PCR like CataCleave probes for SNPs. We continue to study the photophysics and proper coupling of DNA components to multilayer metal nanoparticles for much faster PCR analysis and the use of FCS (see MPM report) to quantify very tight protein-protein and protein-DNA binding in sub-microliter drops (analytes are present in sub-femtomole amounts). We temporarily suspended (this year) doing MPM-FCS and Time-Resolved Fluorescence together; in previous years, we had numerically combined time-resolved fluorescence detection with translational mobility (FCS) to help identify free and bound signatures for assay. We are presently designing STAQ probes (see nanoscopy project) for DNA/RNA probing, analagous to catacleave, in plans to only superresolve tight binding sites. Recently we have used TCSPC to begin untangling designed aptamer heterogeneity questions (see MPM project and TR project). This year, we also began studying Excitonic probes that self-quench unless unfolded or cleaved, in collaboration with industry.

Project Start
Project End
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Budget End
Support Year
9
Fiscal Year
2018
Total Cost
Indirect Cost
Name
U.S. National Heart Lung and Blood Inst
Department
Type
DUNS #
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Dasgeb, Bahar; Smirnov, Aleksandr V; Ardeshirpour, Yasaman et al. (2014) Multiscale BerEp4 molecular imaging of microtumor phantoms: toward theranostics for basal cell carcinoma. Mol Imaging 13:
Ardeshirpour, Yasaman; Chernomordik, Victor; Zielinski, Rafal et al. (2012) In vivo fluorescence lifetime imaging monitors binding of specific probes to cancer biomarkers. PLoS One 7:e31881
Ben-Aissa, Khadija; Patino-Lopez, Genaro; Belkina, Natalya V et al. (2012) Activation of moesin, a protein that links actin cytoskeleton to the plasma membrane, occurs by phosphatidylinositol 4,5-bisphosphate (PIP2) binding sequentially to two sites and releasing an autoinhibitory linker. J Biol Chem 287:16311-23
Fiammenghi, Laura; Ancarani, Valentina; Rosales, Tilman et al. (2010) FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy. J Transl Med 8:52
Wojtuszewski Poulin, Kristi; Smirnov, Aleksandr V; Hawkins, Mary E et al. (2009) Conformational heterogeneity and quasi-static self-quenching in DNA containing a fluorescent guanine analogue, 3MI or 6MI. Biochemistry 48:8861-8