Abstract

BackgroundSingle particle tracking (SPT) is nowadays one of the most popular technique to probe spatio-temporal dynamics of proteins diffusing within the plasma membrane. Indeed membrane components of eukaryotic cells are very dynamic molecules and can diffuse according to different motion modes. Trajectories are often reconstructed frame-by-frame and dynamic properties often evaluated using mean square displacement (MSD) analysis. However, to get statistically significant results in tracking experiments, analysis of a large number of trajectories is required and new methods facilitating this analysis are still needed.ResultsIn this study we developed a new algorithm based on back-propagation neural network (BPNN) and MSD analysis using a sliding window. The neural network was trained and cross validated with short synthetic trajectories. For simulated and experimental data, the algorithm was shown to accurately discriminate between Brownian, confined and directed diffusion modes within one trajectory, the 3 main of diffusion encountered for proteins diffusing within biological membranes. It does not require a minimum number of observed particle displacements within the trajectory to infer the presence of multiple motion states. The size of the sliding window was small enough to measure local behavior and to detect switches between different diffusion modes for segments as short as 20 frames. It also provides quantitative information from each segment of these trajectories. Besides its ability to detect switches between 3 modes of diffusion, this algorithm is able to analyze simultaneously hundreds of trajectories with a short computational time.ConclusionThis new algorithm, implemented in powerful and handy software, provides a new conceptual and versatile tool, to accurately analyze the dynamic behavior of membrane components.Electronic supplementary materialThe online version of this article (doi:10.1186/s12859-016-1064-z) contains supplementary material, which is available to authorized users.

Highlights

  • Single particle tracking (SPT) is nowadays one of the most popular technique to probe spatiotemporal dynamics of proteins diffusing within the plasma membrane

  • A combination of confined and Brownian diffusion modes within a trajectory has been observed for transmembrane proteins such as tetraspanins [5], the epidermal growth factor (EGF) receptor [8] and the cystic fibrosis transmembrane conductance regulator (CFTR) channel [9]

  • In this paper we present a new approach to automatically discriminate between different modes of membrane diffusion within the same trajectory using backpropagation neural network (BPNN)

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Summary

Introduction

Single particle tracking (SPT) is nowadays one of the most popular technique to probe spatiotemporal dynamics of proteins diffusing within the plasma membrane. Membrane components of eukaryotic cells are very dynamic molecules and can diffuse according to different motion modes. In addition numerous studies have highlighted that single molecules could present complex behavior, namely switching between the different modes of diffusion described above. It was early described by Jacobson’s group that the glycosylphosphatidylinositol (GPI)-anchored protein Thy could be transiently confined (here alternating between free diffusion and confinement) in specific areas identified as raft microdomains [6]. A combination of confined and Brownian diffusion modes within a trajectory has been observed for transmembrane proteins such as tetraspanins [5], the epidermal growth factor (EGF) receptor [8] and the cystic fibrosis transmembrane conductance regulator (CFTR) channel [9]. Transient directed motion was observed for gamma amino butyric acid (GABA) receptors in nerve growth cones [10]

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