Real-time particle tracking for studying intracellular trafficking of pharmaceutical nanocarriers.
Real-time particle tracking for studying intracellular trafficking of pharmaceutical nanocarriers.
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DOI：
10.1007/978-1-62703-336-7_20
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发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Suh, Junghae
中科院分区：
文献类型：
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作者：
Huang, Feiran;Watson, Erin;Suh, Junghae
实时粒子跟踪是一种将荧光显微镜与物体跟踪和计算相结合的技术，可用于提取细胞内小粒子的定量传输参数。由于纳米载体的成功通常取决于它如何有效地将货物运送到靶细胞器，因此了解药物纳米载体的复杂细胞内转运至关重要。实时粒子跟踪提供了对纳米粒子的细胞内行为的动力学的洞察，这可能会导致新的递送系统的设计和开发的显着改进。不幸的是，这项技术往往没有得到充分理解，限制了纳米医学领域研究人员的实施。在本章中，粒子跟踪最复杂的方面之一，均方位移（MSD）的计算，解释了一个简单的方式设计的新手粒子跟踪器。还提供了在MATLAB中执行MSD计算的伪代码。本章对粒子跟踪技术中的一系列基本步骤进行了清晰而全面的说明。提供了用于执行纳米颗粒样品的共聚焦显微镜的说明，并且提供了两种不需要商业颗粒跟踪软件的确定颗粒轨迹的方法。还说明了轨迹分析和跟踪分辨率的确定。通过提供执行粒子跟踪实验所需的全面说明，本章将使研究人员能够对纳米载体的细胞内动力学获得新的见解，从而可能导致更有效和更智能的治疗递送载体的开发。
Real-time particle tracking is a technique that combines fluorescence microscopy with object tracking and computing and can be used to extract quantitative transport parameters for small particles inside cells. Since the success of a nanocarrier can often be determined by how effectively it delivers cargo to the target organelle, understanding the complex intracellular transport of pharmaceutical nanocarriers is critical. Real-time particle tracking provides insight into the dynamics of the intracellular behavior of nanoparticles, which may lead to significant improvements in the design and development of novel delivery systems. Unfortunately, this technique is not often fully understood, limiting its implementation by researchers in the field of nanomedicine. In this chapter, one of the most complicated aspects of particle tracking, the mean square displacement (MSD) calculation, is explained in a simple manner designed for the novice particle tracker. Pseudo code for performing the MSD calculation in MATLAB is also provided. This chapter contains clear and comprehensive instructions for a series of basic procedures in the technique of particle tracking. Instructions for performing confocal microscopy of nanoparticle samples are provided, and two methods of determining particle trajectories that do not require commercial particle-tracking software are provided. Trajectory analysis and determination of the tracking resolution are also explained. By providing comprehensive instructions needed to perform particle-tracking experiments, this chapter will enable researchers to gain new insight into the intracellular dynamics of nanocarriers, potentially leading to the development of more effective and intelligent therapeutic delivery vectors.