Spatial Structure and Diffusive Dynamics from Single Particle Trajectories Using Spline Analysis英文文献.pdfVIP

1712 Biophysical Journal Volume 98 April 2010 1712–1721 Spatial Structure and Diffusive Dynamics from Single-Particle Trajectories Using Spline Analysis Brian R. Long and Tania Q. Vu* Biomedical Engineering, Oregon Health Science University, Portland, Oregon ABSTRACT Single-particle tracking of biomolecular probes has provided a wealth of information about intracellular trafficking and the dynamics of proteins and lipids in the cell membrane. Conventional mean-square displacement (MSD) analysis of single- particle trajectories often assumes that probes are moving in a uniform environment. However, the observed two-dimensional motion of probe particles is influenced by the local three-dimensional geometry of the cell membrane and intracellular structures, which are rarely flat at the submicron scale. This complex geometry can lead to spatially confined trajectories that are difficult to analyze and interpret using conventional two-dimensional MSD analysis. Here we present two methods to analyze spatially confined trajectories: spline-curve dynamics analysis, which extends conventional MSD analysis to measure diffusive motion in confined trajectories; and spline-curve spatial analysis, which measures spatial structures smaller than the limits of optical resolution. We show, using simulated random walks and experimental trajectories of quantum dot probes, that differences in measured two-dimensional diffusion coefficients do not always reflect differences in underlying diffusive dynamics, but can instead be due to differences in confinement geometries of cellular structures. INTRODUCTION Single-particle tracking (SPT) is becoming widely applied in (22–25). To calculate the MSD from a trajectory, the the life sciences, including the use of microscopy to track displacements over the time difference Dt a