Measuring Stepwise Binding of Thermally Fluctuating Particles to Cell Membranes without Fluorescence

Titelangaben

Rohrbach, Alexander ; Meyer, Tim ; Stelzer, Ernst H. K. ; Kress, Holger:
Measuring Stepwise Binding of Thermally Fluctuating Particles to Cell Membranes without Fluorescence.
In: Biophysical Journal. Bd. 118 (2020) Heft 8 . - S. 1850-1860.
ISSN 1542-0086
DOI: https://doi.org/10.1016/j.bpj.2020.03.005

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Abstract

Thermal motions enable a particle to probe the optimal interaction state when binding to a cell membrane. However, especially on the scale of microseconds and nanometers, position and orientation fluctuations are difficult to observe with common measurement technologies. Here, we show that it is possible to detect single binding events of immunoglobulin-G-coated polystyrene beads, which are held in an optical trap near the cell membrane of a macrophage. Changes in the spatial and temporal thermal fluctuations of the particle were measured interferometrically, and no fluorophore labeling was required. We demonstrate both by Brownian dynamic simulations and by experiments that sequential stepwise increases in the force constant of the bond between a bead and a cell of typically 20 pN/μm are clearly detectable. In addition, this technique provides estimates about binding rates and diffusion constants of membrane receptors. The simple approach of thermal noise tracking points out new strategies in understanding interactions between cells and particles, which are relevant for a large variety of processes, including phagocytosis, drug delivery, and the effects of small microplastics and particulates on cells.