A Platform for Fast Detection of Let-7 Micro RNA Using Polyaniline Fluorescence and Image Analysis Techniques
The project describes a new strategy for transducing hybridization events through modulating intrinsic properties of the electroconductive polymer polyaniline (PANI). When DNA based probes electrostatically interact with PANI, its fluorescence properties are increased, a phenomenon that can be enhanced by UV irradiation. Hybridization of target nucleic acids results in dissociation of probes causing PANI fluorescence to return to basal levels. By monitoring restoration of base PANI fluorescence as little as 10-11 M (10 pM) of target oligonucleotides could be detected within 15 minutes of hybridization. Detection of complementary oligos was specific, with introduction of a single mismatch failing to form a target-probe duplex that would dissociate from PANI. Furthermore, this approach is robust and is capable of detecting specific RNAs in extracts from animals. This sensor system improves on previously reported strategies by transducing highly specific probe dissociation events through intrinsic properties of a conducting polymer without the need for additional labels. The change in fluorescence property of PANI by oligo immobilization and hybridization with mimic let-7 is measured by fluorescence microscope and the image analyzed by MATLAB. A heuristic algorithm determines color threshold of the fluorescent active image. This image segmentation helps to determine the average pixel intensity representing the active image foreground of PANI fluorescence triggered by DNA immobilization and hybridization process. This would help us to quantify response of PANI based biosensor for detecting micro RNA let-7.
micro RNA, Polyaniline, electrostatic interaction, fluorescence, UV irradiation, intrinsic property, Biology and Biomimetic Materials, Computer Engineering, Genetics and Genomics, Polymer and Organic Materials, Semiconductor and Optical Materials
Networked Digital Library of Theses and Dissertations