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 previously 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 numerically combined time-resolved fluorescence detection with translational mobility (FCS) to help identify """"""""free"""""""" and """"""""bound"""""""" signatures for assay. We have just recently installed a RICS /FCCS microscope for combining the techniques mentioned above.

Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2013
Total Cost
$47,622
Indirect Cost
Name
National Heart, Lung, and Blood Institute
Department
Type
DUNS #
City
State
Country
Zip Code
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