Membrane models have allowed for precise study of the plasma membrane's biophysical properties, helping to unravel both structural and dynamic motifs within cell biology. Freestanding and supported bilayer systems are popular models to reconstitute membrane-related processes. Although it is wellknown that each have their advantages and limitations, comprehensive comparison of their biophysical properties is still lacking. Here, we compare the diffusion and lipid packing in giant unilamellar vesicles, planar and spherical supported membranes, and cell-derived giant plasma membrane vesicles. We apply florescence correlation spectroscopy (FCS), spectral imaging, and super-resolution stimulated emission depletion FCS to study the diffusivity, lipid packing, and nanoscale architecture of these membrane systems, respectively. Our data show that lipid packing and diffusivity is tightly correlated in freestanding bilayers. However, nanoscale interactions in the supported bilayers cause deviation from this correlation. These data are essential to develop accurate theoretical models of the plasma membrane and will serve as a guideline for suitable model selection in future studies to reconstitute biological processes.

Understanding the broad variety of functions encoded in cellular membranes requires experimental systems mimicking both their biochemical composition and biophysical properties. Here, we review the interplay between membrane components and the physical properties of the plasma membrane worth considering for biomimetic studies. Later, we discuss the main advantages and caveats of different model membrane systems. We further expand on how the use of model systems has contributed to the understanding of immune cell signaling, with a specific focus on the immunological synapse. We discuss the similarities of immune synapses observed for innate and adaptive immune cells and focus on the physical principles underlying these similarities.

Membrane models have allowed for precise study of the plasma membrane's biophysical properties, helping to unravel both structural and dynamic motifs within cell biology. Free standing and supported bilayer systems are popular models to reconstitute the membrane related processes. Although it is well-known that each have their advantages and limitations, comprehensive comparison of their biophysical properties is still lacking. Here, we compare the diffusion and lipid packing in giant unilamellar vesicles, planar and spherical supported membranes and cell-derived giant plasma membrane vesicles. We apply florescence correlation spectroscopy, spectral imaging and super-resolution STED-FCS to study the diffusivity, lipid packing and nanoscale architecture of these membrane systems, respectively.Our data show that lipid packing and diffusivity is tightly correlated in free-standing bilayers.However, nanoscale interactions in the supported bilayers cause deviation from this correlation.This data is essential to develop accurate theoretical models of the plasma membrane and will serve as a guideline for suitable model selection in future studies to reconstitute biological processes.