title: A quantitative comparison of single-dye tracking analysis tools using Monte Carlo simulations.
abstract: Single-particle tracking (SPT) is widely used to study processes from membrane receptor organization to the dynamics of RNAs in living cells. While single-dye labeling strategies have the benefit of being minimally invasive, this comes at the expense of data quality; typically a data set of short trajectories is obtained and analyzed by means of the mean square displacements (MSD) or the distribution of the particles' displacements in a set time interval (jump distance, JD). To evaluate the applicability of both approaches, a quantitative comparison of both methods under typically encountered experimental conditions is necessary. Here we use Monte Carlo simulations to systematically compare the accuracy of diffusion coefficients (D-values) obtained for three cases: one population of diffusing species, two populations with different D-values, and a population switching between two D-values. For the first case we find that the MSD gives more or equally accurate results than the JD analysis (relative errors of D-values <6%). If two diffusing species are present or a particle undergoes a motion change, the JD analysis successfully distinguishes both species (relative error <5%). Finally we apply the JD analysis to investigate the motion of endogenous LPS receptors in live macrophages before and after treatment with methyl-β-cyclodextrin and latrunculin B.
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge, United Kingdom.
authors.forename: Laura
authors.initials: L
authors.lastname: Weimann
authors.forename: Kristina A
authors.initials: KA
authors.lastname: Ganzinger
authors.forename: James
authors.initials: J
authors.lastname: McColl
authors.forename: Kate L
authors.initials: KL
authors.lastname: Irvine
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Davis
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Gay
authors.forename: Clare E
authors.initials: CE
authors.lastname: Bryant
authors.forename: David
authors.initials: D
authors.lastname: Klenerman
mesh_terms: D000465:Algorithms; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D004396:Coloring Agents / Q000378:metabolism*; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D051379:Mice; D015336:Molecular Probe Techniques; D009010:Monte Carlo Method; D051197:Toll-Like Receptor 4 / Q000378:metabolism
chemical_list: D004396:Coloring Agents; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D051197:Toll-Like Receptor 4