Analysing Sources of Error in Total Internal Reflection Microscopy (TIRM) Experiments and Data Analysis

Abstract

Many phenomena observed in synthetic and biological colloidal suspensions are dominated by the static interaction energies and the hydrodynamic interactions that act both between individual particles and also between colloids and macroscopic interfaces. This calls for methods that allow precise measurements of the corresponding forces. One method used for this purpose is total internal reflection microscopy (TIRM), which has been employed for around three decades to measure in particular the interactions between a single particle suspended in a liquid and a solid surface. However, given the importance of the observable variables, it is crucial to understand the possibilities and limitations of the method. In this paper, we investigate the influence of technically unavoidable noise effects and an inappropriate choice of particle size and sampling time on TIRM measurement results. Our main focus is on the measurement of diffusion coefficients and drift velocities, as the influence of error sources on dynamic properties has not been investigated so far. We find that detector shot noise and prolonged sampling times may cause erroneous results in the steep parts of the interaction potential where forces of the order of pico-Newtons or larger act on the particle, while the effect of background noise is negligible below certain thresholds. Furthermore, noise does not significantly affect dynamic data but we find that lengthy sampling times and/or probe particles with too small a radius will cause issues. Most importantly, we observe that dynamic results are very likely to differ from the standard hydrodynamic predictions for stick boundary conditions due to partial slip.