Dengue Virus Capsid-Protein Dynamics in Live Infected Cells Studied by Pair Correlation Analysis.
Dengue Virus Capsid-Protein Dynamics in Live Infected Cells Studied by Pair Correlation Analysis.
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DOI：
10.1007/978-1-0716-1879-0_8
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发表时间：
2021-10
影响因子：
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通讯作者：
M. Gabriel;Ignacio Sallaberry;Guadalupe S. Costa Navarro;E. Gratton;A. Gamarnik;L. Estrada
中科院分区：
文献类型：
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作者：
M. Gabriel;Ignacio Sallaberry;Guadalupe S. Costa Navarro;E. Gratton;A. Gamarnik;L. Estrada
越来越明显的是，揭示病毒进入、组装和病毒体释放的机制对于确定预防病毒传播和控制病毒性疾病的方法是至关重要的。由于病毒的流动性和病毒颗粒之间的结构和/或功能的异质性，需要高时空分辨率的单病毒/单颗粒技术来捕捉病毒颗粒在感染细胞内的行为，在本章中，我们提出了荧光成像分析方法来研究荧光标记的登革病毒（DENV）蛋白在活感染细胞中的流动性。一些最新的荧光波动光谱（FFS）的方法将被提出，特别是，对相关函数（pCF）的方法将被讨论。pCF方法不需要单独的分子分离，如在颗粒跟踪实验中，以捕获单个病毒蛋白的行为。在这方面，图像采集之后是时空互相关函数在增加的时间延迟，产生一个定量的观点，在完整的活感染cells.We单粒子的流动性提供了一个一般性的概述和先进的FFS的实施的实际指导 技术和配对相关函数分析，作为定量工具，以揭示以前未报告的DENV的见解 机制等我们希望这份报告将作为进一步应用相关成像研究在病毒学研究的激励。
It has become increasingly evident that unveiling the mechanisms of virus entry, assembly, and virion release is fundamental for identifying means for preventing viral spread and controlling viral disease. Due to virus mobility and structural and/or functional heterogeneity among viral particles, high spatiotemporal resolution single-virus/single-particle techniques are required to capture the behavior of viral particles inside infected cells.In this chapter, we present fluorescence imaging analysis methods for studying the mobility of fluorescently labeled dengue virus (DENV) proteins in live infected cells. Some of the most recent Fluorescence Fluctuation Spectroscopy (FFS) methods will be presented and, in particular, the pair Correlation Functions (pCF) approach will be discussed. The pCF method does not require individual molecule isolation, as in a particle-tracking experiment, to capture single viral protein behavior. In this regard, image acquisition is followed by the spatiotemporal cross-correlation function at increasing time delays, yielding a quantitative view of single-particle mobility in intact live infected cells.We provide a general overview and a practical guidance for the implementation of advanced FFS techniques, and the pair Correlation Functions analysis, as quantitative tools to reveal insights into previously unreported DENV mechanisms. We expect this protocol report will serve as an incentive for further applying correlation imaging studies in virology research.