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This paper presents the alkanethiol-functionalized environmental effects of individual DNA molecules in nanopores on enzyme digestion at the single-molecule level. A template consisting of gold deposited within a solid-state nanoporous polycarbonate membrane was used to trap individual λ-DNA and enzyme molecules. The gold surfaces were modified with various functional groups (–OH, –COOH, –NH 3...
Total internal reflection fluorescence microscopy (TIRFM) was investigated for enhanced detection sensitivity of cardiac troponin I (cTnI) on a gold nano-patterned protein chip. A 4×5 nanoarray patterning of a gold chip based on a single-molecule sandwich immunoassay was prepared with a spot diameter of 500nm on glass substrates formed via electron-beam nanolithography. Quantitative trace analysis...
We performed wide-range quantification of human thyroid-stimulating hormone (hTSH) using a gold nano-patterned sandwich immunoassay chip. Objective-type total internal reflection fluorescence microscopy (TIRFM) was used to detect hTSH at the single-molecule level. A gold spot with a diameter of 100nm on a 10-mm square glass substrate was fabricated by electron beam nanolithography. When hTSH bound...
The real-time digestion of entrapped single-DNA molecules by λ-exonuclease in nanoporous alumina membranes was observed using an epifluorescence microscope. The alumina membrane provides pL (∼10 −12 L) containers for confining single-DNA molecules without immobilization. When one end of the DNA molecule was inserted into a nanopore, it was possible to monitor the digestion process outside,...
We report a method for the quantitative analysis of human serum leptin, which is a protein hormone associated with obesity, using a nanoarray protein chip based on a single-molecule sandwich immunoassay. The nanoarray patterning of a biotin-probe with a spot diameter of 150nm on a self-assembled monolayer functionalized by MPTMS on a glass substrate was successfully accomplished using atomic force...
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