Phagocytosis, the uptake and ingestion of solid particles into living cells, is a central mechanism of our immune system. Due to the complexity of the uptake mechanism, the different forces involved in this process are only partly understood. Therefore the usage of a giant unilamellar vesicle (GUV) as the simplest biomimetic model for a cell allows one to investigate the influence of the lipid membrane on the energetics of the uptake process. Here, a photonic force microscope (PFM) is used to approach an optically trapped 1 μm latex bead to an immobilized GUV to finally insert the particle into the GUV. By analysing the mean displacement and the position fluctuations of the trapped particle during the uptake process in 3D with nanometre precision, we are able to record force and energy profiles, as well as changes in the viscous drag and the stiffness. After observing a global followed by a local deformation of the GUV, we measured uptake energies of 2000 kT to 5500 kT and uptake forces of 4 pN to 16 pN for Egg-PC GUVs with sizes of 18-26 μm and varying membrane tension. The measured energy profiles, which are compared to a Helfrich energy model for local and global deformation, show good coincidence with the theoretical results. Our proof-of-principle study opens the door to a large number of similar experiments with GUVs containing more biochemical components and complexity. This bottom-up strategy should allow for a better understanding of the physics of phagocytosis.

Multi-dimensional, correlated particle tracking is a key technology to reveal dynamic processes in living and synthetic soft matter systems. In this paper we present a new method for tracking micron-sized beads in parallel and in all three dimensions - faster and more precise than existing techniques. Using an acousto-optic deflector and two quadrant-photo-diodes, we can track numerous optically trapped beads at up to tens of kHz with a precision of a few nanometers by back-focal plane interferometry. By time-multiplexing the laser focus, we can calibrate individually all traps and all tracking signals in a few seconds and in 3D. We show 3D histograms and calibration constants for nine beads in a quadratic arrangement, although trapping and tracking is easily possible for more beads also in arbitrary 2D arrangements. As an application, we investigate the hydrodynamic coupling and diffusion anomalies of spheres trapped in a 3 × 3 arrangement.

The interaction of two diffusing particles is strongly influenced by their hydrodynamic coupling. At a tracking rate of 10 kHz we are able to measure the 3D trajectories of two colloidal spheres in a single harmonic potential, which was generated by scanning line optical tweezers. This common potential enables tilting, rotational, and translational dynamics of the spheres, which we analyzed via the spheres position cross-correlations C(τ) over a time range of 10(-4)-2 s. We found that the dynamic interaction of the colloids is controlled by short-range surface forces F(s), which are attractive in one direction and repulsive in the other two directions. This unexpected behavior is supported by a theoretical model using two Langevin equations, which decouple for linear F(s), allowing a description with autocorrelation functions for collective and relative motions. We further demonstrate that variations in salt concentration and reaction volumes significantly influence C(τ) and the mean contact times between the particles, which may offer new insights into biological particle interaction.

Interactions of diffusing particles are governed by hydrodynamics on different length and timescales. The local hydrodynamics can be influenced substantially by simple interfaces. Here, we investigate the interaction dynamics of two micron-sized spheres close to plane interfaces to mimic more complex biological systems or microfluidic environments. Using scanned line optical tweezers and fast 3D interferometric particle tracking, we are able to track the motion of each bead with precisions of a few nanometers and at a rate of 10 kilohertz. From the recorded trajectories, all spatial and temporal information is accessible. This way, we measure diffusion coefficients for two coupling particles at varying distances h to one or two glass interfaces. We analyze their coupling strength and length by cross-correlation analysis relative to h and find a significant decrease in the coupling length when a second particle diffuses nearby. By analysing the times the particles are in close contact, we find that the influence of nearby surfaces and interaction potentials reduce the diffusivity strongly, although we found that the diffusivity hardly affects the contact times and the binding probability between the particles. All experimental results are compared to a theoretical model, which is based on the number of possible diffusion paths following the Catalan numbers and a diffusion probability, which is biased by the spheres' surface potential. The theoretical and experimental results agree very well and therefore enable a better understanding of hydrodynamically coupled interaction processes.