论文标题
用于高吞吐量单分子纳米纳米动力学的多路复用等离激元纳米antennas在活细胞中
Multiplexed plasmonic nanoantennas for high throughput single molecule nanoscale dynamics in living cells
论文作者
论文摘要
单分子检测通过在不同情况下研究单个分子的强大方法来彻底改变化学和生物学领域。包括等离子体天线在内的纳米光结构可显着克服可以观察到单分子事件的浓度极限,从而使其在与生物学和化学过程有关的浓度下进行检测。尽管天线可以在大型阵列中制造,但探测动态事件需要高的时间分辨率,这是通过串行天线询问来实现的。不幸的是,这排除了不同样本位置的多个天线的同时记录,并且耗时,导致统计数据差和低通量数据获取,从而减少了阵列的真正潜力。在这里,我们将天线内部纳米结构的阵列与SCMOS读数结合使用,以同时从225个天线询问纳米级体积。在1 kHz处记录允许同时从50纳米纳斯的多路复用动态测量,并在单个通道期间由摄像机帧帧指示的时间分辨率和每个分子发出的光子。我们证明了活细胞膜上纳米级对单分子动力学的高通量阵列检测的能力,并确定膜分子成分的轴向位置,其精度为1 nm,时间分辨率低于1 ms。
Single molecule detection has revolutionised the fields of chemistry and biology by offering powerful ways to study individual molecules under different scenarios. Nanophotonic structures, including plasmonic antennas, significantly overcome the concentration limit at which single molecule events can be observed, enabling their detection at concentrations that are relevant to biological and chemical processes. Although antennas can be fabricated in large arrays, probing dynamic events requires high temporal resolution, which is best achieved by serial antenna interrogation. Unfortunately, this precludes the simultaneous recording from multiple antennas at different sample locations, and is time consuming, resulting in poor statistics and low-throughput data acquisition, abating the true potential of arrays. Here we exploit arrays of antenna-in-box nanostructures in combination with sCMOS readout to interrogate nanoscale volumes from 225 antennas simultaneously. Recording at 1 kHz allowed multiplexed dynamic measurements from 50 nanoantennas simultaneously with a temporal resolution dictated by the camera framerate and the photons emitted per molecule during a single passage. We demonstrate the capability for high-throughput arrayed detection of single molecule dynamics at the nanoscale in the membrane of living cells, and determine the axial location of membrane molecular components with 1 nm accuracy and temporal resolution below 1 ms.