Two Color DNA Barcode Detection in Photoluminescence Suppressed Silicon Nitride Nanopores

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• 作者：Ossama N. Assad ; Nicolas Di Fiori ; Allison H. Squires ; Amit Meller
• 刊名：Nano Letters
• 出版年：2015
• 出版时间：January 14, 2015
• 年：2015
• 卷：15
• 期：1
• 页码：745-752
• 全文大小：375K
• ISSN：1530-6992

文摘

Optical sensing of solid-state nanopores is a relatively new approach that can enable high-throughput, multicolor readout from a collection of nanopores. It is therefore highly attractive for applications such as nanopore-based DNA sequencing and genotyping using DNA barcodes. However, to date optical readout has been plagued by the need to achieve sufficiently high signal-to-noise ratio (SNR) for single fluorophore sensing, while still maintaining millisecond resolution. One of the main factors degrading the optical SNR in solid-state nanopores is the high photoluminescence (PL) background emanating from the silicon nitride (SiN_x) membrane in which pores are commonly fabricated. Focusing on the optical properties of SiN_x nanopores we show that the local membrane PL intensity is substantially reduced, and its spectrum is shifted toward shorter wavelengths with increasing e-beam dose. This phenomenon, which is correlated with a marked photocurrent enhancement in these nanopores, is utilized to perform for the first time single molecule fluorescence detection using both green and red laser excitations. Specifically, the reduction in PL and the concurrent measurement of the nanopore photocurrent enhancement allow us to maximize the background suppression and to detect a dual color, five-unit DNA barcode with high SNR levels.