Abstract
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.
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References
Suh J et al (2003) Efficient active transport of gene nanocarriers to the cell nucleus. Proc Natl Acad Sci USA 100:3878–3882
Huang F et al (2011) Quantitative nanoparticle tracking: applications to nanomedicine. Nanomedicine 6(4):693–700
Qian H et al (1991) Single particle tracking. Analysis of diffusion and flow in two-dimensional systems. Biophys J 60:910–921
Kusumi A et al (1993) Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells. Biophys J 65:2021–2040
Saxton MJ, Jacobson K (1997) Single-particle tracking: applications to membrane dynamics. Annu Rev Biophys Biomol Struct 26:373–399
Suh J et al (2005) Real-time multiple-particle tracking: applications to drug and gene delivery. Adv Drug Deliv Rev 57:63–78
Lai SK, Hanes J (2008) Real-time multiple particle tracking of gene nanocarriers in complex biological environments. Methods Mol Biol 434:81–97
Kawai M et al (2009) Dynamics of different-sized solid-state nanocrystals as tracers for a drug-delivery system in the interstitium of a human tumor xenograft. Breast Cancer Res 11:R43
Chen C, Suh J (2010) Real-time particle tracking for studying intracellular transport of nanotherapeutics. In: Weissig V, D’Souza GGM (eds) Organelle-specific pharmaceutical nanotechnology. Wiley, Hoboken
Wirtz D (2009) Particle-tracking microrheology of living cells: principles and applications. Annu Rev Biophys 38:301–326
Sage D et al (2005) Automatic tracking of individual fluorescence particles: application to the study of chromosome dynamics. IEEE Trans Image Process 14:1372–1383
Acknowledgment
This work was supported by the National Institutes of Health (RC2GM092599 and T32EB009379).
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Huang, F., Watson, E., Dempsey, C., Suh, J. (2013). Real-Time Particle Tracking for Studying Intracellular Trafficking of Pharmaceutical Nanocarriers. In: Weissig, V., Elbayoumi, T., Olsen, M. (eds) Cellular and Subcellular Nanotechnology. Methods in Molecular Biology, vol 991. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-336-7_20
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DOI: https://doi.org/10.1007/978-1-62703-336-7_20
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Publisher Name: Humana Press, Totowa, NJ
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