Nanofluidic Devices with 8 Pores in Series for Real-Time, Resistive-Pulse Analysis of Hepatitis B Virus Capsid Assembly.
Nanofluidic Devices with 8 Pores in Series for Real-Time, Resistive-Pulse Analysis of Hepatitis B Virus Capsid Assembly.
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DOI：
10.1021/acs.analchem.6b04491
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发表时间：
2017-05-02
影响因子：
7.4
通讯作者：
Jacobson SC
中科院分区：
文献类型：
--
作者：
Kondylis P;Zhou J;Harms ZD;Kneller AR;Lee LS;Zlotnick A;Jacobson SC
为了提高连续脉冲测量的精度，我们使用聚焦离子束仪器来研磨具有2个、4个和8个串联孔的纳米流体装置，并比较它们的性能。面内设计有助于连续制造多个孔，这反过来又允许对每个移位事件产生的一系列脉冲进行平均。当将八个脉冲的平均幅度与单个脉冲的幅度进行比较时，脉冲幅度分布的标准偏差（σ）减小了2.7倍。类似地，当将8孔装置的7次测量的平均值与2孔装置的单次测量值进行对比时，孔-孔时间分布的标准偏差降低了3.2倍。通过信号平均，测量中的固有不确定性降低;因此，分辨率（平均值/σ）提高了一个等于测量次数平方根的因子。我们利用8孔装置的改进的尺寸分辨率来真实的实时分析低于伪临界浓度的B型肝炎病毒（HBV）衣壳的组装。我们观察到组装中间体的丰度随时间而变化。在反应的第一个小时期间，较小的中间体的丰度降低，而大小接近T = 4衣壳的较大中间体的丰度保持恒定。
To improve the precision of resistive-pulse measurements, we have used a focused ion beam instrument to mill nanofluidic devices with 2, 4, and 8 pores in series and compared their performance. The in-plane design facilitates the fabrication of multiple pores in series, which, in turn, permits averaging of the series of pulses generated from each translocation event. The standard deviations (σ) of the pulse amplitude distributions decrease by 2.7-fold when the average amplitudes of eight pulses are compared to the amplitudes of single pulses. Similarly, standard deviations of the pore-to-pore time distributions decrease by 3.2-fold when the averages of the seven measurements from 8-pore devices are contrasted to single measurements from 2-pore devices. With signal averaging, the inherent uncertainty in the measurements decreases; consequently, the resolution (mean/σ) improves by a factor equal to the square root of the number of measurements. We took advantage of the improved size resolution of the 8-pore devices to analyze in real time the assembly of Hepatitis B Virus (HBV) capsids below the pseudo-critical concentration. We observe that abundances of assembly intermediates change over time. During the first hour of the reaction, the abundance of smaller intermediates decreased, whereas the abundance of larger intermediates with sizes closer to a T = 4 capsid remained constant.
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影响因子：
15
作者：
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通讯作者：
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