High-throughput single molecule screening of DNA and proteins.
We report a novel imaging technology for real time comprehensive analysis of molecular alterations in cells and tissues appropriate for automation and adaptation to high-throughput applications. With these techniques it should eventually be possible to perform simultaneous analysis of the entire contents of individual biological cells with a sensitivity and selectivity sufficient to determine the presence or absence of a single copy of a targeted analyte (e.g., DNA region, RNA region, protein), and to do so at a relatively low cost. The technology is suitable for DNA and RNA through sizing or through fluorescent hybridization probes, and for proteins and small molecules through fluorescence immunoassays. This combination of the lowest possible detection limit and the broadest applicability to biomolecules represents the final frontier in bioanalysis. The general scheme is based on novel concepts for single molecule detection (SMD) and characterization recently demonstrated in our laboratory. Since minimal manipulation is involved, it should be possible to screen large numbers of cells in a short time to facilitate practical applications. This opens up the possibility of finding single copies of DNA or proteins within single biological cells for disease markers without performing polymerase chain reaction or other biological amplification.
Yeung
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Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, 50011, USA. yeung@ameslab.gov
D000818:Animals; D001704:Biopolymers / Q000032:analysis / Q000737:chemistry; D002469:Cell Separation / Q000379:methods; D002477:Cells / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D004058:Diffusion; D004586:Electrophoresis / Q000379:methods; D005453:Fluorescence; D006801:Humans; D036103:Nanotechnology / Q000379:methods; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D012313:RNA / Q000032:analysis / Q000737:chemistry
D001704:Biopolymers; D011506:Proteins; D012313:RNA; D004247:DNA