Fluorescence Spectroscopy is inherently a very sensitive technique; it already forms the basis of most non-radioactive real time assays like PCR. Our lab has collaborated with former fellows (now in biotech industry) to develop alternatives to PCR like CataCleave probes for SNPs, leading to previous publication. 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 have examined the structural transitions of similar amounts of DNA between G-quadruplexed and linear forms, using both MPM-FCS and Time-Resolved Fluorescence tools, with the goal of developing very sensitive 'G-quad' and aptamer detection assays. We have begun to translate aptamer experience into BCC/melanoma detection (see MPM project)collaboration. We have numerically combined time-resolved fluorescence detection with translational mobility (FCS) to help identify free and bound signatures for assay. We have a RICS /FCCS microscope which is available part time for combining the techniques mentioned above. We are also designing STAQ probes (see nanoscopy project) for DNA/RNA probing, analagous to catacleave.

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Budget End
Support Year
6
Fiscal Year
2015
Total Cost
Indirect Cost
Name
U.S. National Heart Lung and Blood Inst
Department
Type
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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:
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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
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