Zhang Miao, Schmidt Torsten, Jemt Anders, Sahlén Pelin, Sychugov Ilya, Lundeberg Joakim, Linnros Jan
Materials and Nano Physics, School of Information and Communication Technology, KTH Royal Institute of Technology, Electrum 229, SE-16440 Kista-Stockholm, Sweden.
Nanotechnology. 2015 Aug 7;26(31):314002. doi: 10.1088/0957-4484/26/31/314002. Epub 2015 Jul 16.
Optical nanopore sensing offers great potential in single-molecule detection, genotyping, or DNA sequencing for high-throughput applications. However, one of the bottle-necks for fluorophore-based biomolecule sensing is the lack of an optically optimized membrane with a large array of nanopores, which has large pore-to-pore distance, small variation in pore size and low background photoluminescence (PL). Here, we demonstrate parallel detection of single-fluorophore-labeled DNA strands (450 bps) translocating through an array of silicon nanopores that fulfills the above-mentioned requirements for optical sensing. The nanopore array was fabricated using electron beam lithography and anisotropic etching followed by electrochemical etching resulting in pore diameters down to ∼7 nm. The DNA translocation measurements were performed in a conventional wide-field microscope tailored for effective background PL control. The individual nanopore diameter was found to have a substantial effect on the translocation velocity, where smaller openings slow the translocation enough for the event to be clearly detectable in the fluorescence. Our results demonstrate that a uniform silicon nanopore array combined with wide-field optical detection is a promising alternative with which to realize massively-parallel single-molecule detection.
光学纳米孔传感在单分子检测、基因分型或用于高通量应用的DNA测序方面具有巨大潜力。然而,基于荧光团的生物分子传感的瓶颈之一是缺乏一种光学优化的膜,该膜具有大量纳米孔,孔间距大、孔径变化小且背景光致发光(PL)低。在这里,我们展示了对单个荧光团标记的DNA链(450个碱基对)通过满足上述光学传感要求的硅纳米孔阵列进行并行检测。纳米孔阵列是通过电子束光刻和各向异性蚀刻,然后进行电化学蚀刻制造的,孔径低至约7纳米。DNA转位测量是在为有效控制背景PL而定制的传统宽视场显微镜中进行的。发现单个纳米孔直径对转位速度有重大影响,较小的孔径会使转位足够慢,从而在荧光中能清晰检测到该事件。我们的结果表明,均匀的硅纳米孔阵列与宽视场光学检测相结合是实现大规模并行单分子检测的一种有前途的替代方法。