Advanced fluorescence microscopy methods illuminate the transfection pathway of nucleic acid nanoparticles
Advanced fluorescence microscopy methods illuminate the transfection pathway of nucleic acid nanoparticles
复制标题
DOI：
10.1016/j.jconrel.2010.08.029
复制
发表时间：
2010-11-20
影响因子：
10.8
通讯作者：
De Smedt, Stefaan C.
中科院分区：
文献类型：
--
作者：
Braeckmans, Kevin;Buyens, Kevin;De Smedt, Stefaan C.
生物制药和制药技术的大量关注集中在纳米颗粒的开发上，以有效地将核酸递送至靶细胞。尽管核酸在治疗各种疾病方面具有巨大潜力，但该领域的进展相当缓慢。原因之一是，由于对转染过程各个阶段的理化和生物物理特性了解不足，阻碍了合适的纳米级递送载体的开发。为了解决这个问题，在过去的十年中，我们开发并应用了先进的荧光显微镜技术，可以更好地了解纳米粒子在各种生物介质中的运输和稳定性。这篇小综述讨论了光漂白后荧光恢复 (FRAP)、荧光相关光谱 (FCS) 和单粒子追踪 (SPT) 的基本原理，并概述了我们使用这些技术来表征细胞外介质和活细胞中含有核酸的纳米颗粒的运输和稳定性的研究。 (C) 2010 Elsevier B.V. 保留所有权利。
A great deal of attention in biopharmacy and pharmaceutical technology is going to the development of nanoscopic particles to efficiently deliver nucleic acids to target cells. Despite the great potential of nucleic acids for treatment of various diseases, progress in the field is fairly slow. One of the causes is that development of suitable nanoscopic delivery vehicles is hampered by insufficient knowledge of their physicochemical and biophysical properties during the various phases of the transfection process. To address this issue, in the past decade we have developed and applied advanced fluorescence microscopy techniques that can provide a better insight in the transport and stability of nanoparticles in various biological media. This mini-review discusses the basic principles of fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS) and single particle tracking (SPT), and gives an overview of studies in which we have employed these techniques to characterize the transport and stability of nucleic acid containing nanoparticles in extracellular media and in living cells. (C) 2010 Elsevier B.V. All rights reserved.