Data and code from: Quantifying 3D live-cell membrane dynamics using dynamic metal-induced energy transfer spectroscopy (dynaMIET)

The dynamic behavior of cellular membranes underpins essential biological
processes, including signal transduction, intracellular trafficking, and
mechanotransduction. However, simultaneously quantifying lateral molecular
diffusion and vertical membrane fluctuations in live cells remains
challenging. Here, we present dynamic metal-induced energy transfer
spectroscopy (dynaMIET), which integrates metal-induced energy transfer
with fluorescence correlation spectroscopy to resolve three-dimensional
membrane dynamics with nanometer axial sensitivity and microsecond
temporal resolution. dynaMIET enables concurrent measurement of lateral
diffusion and vertical undulations within a single acquisition. We
validate the method using simulations and model membranes and demonstrate
its robustness in living cells, applying it to the plasma membrane,
endoplasmic reticulum, and nuclear envelope. By capturing both molecular
mobility and membrane fluctuations, dynaMIET provides a powerful,
non-invasive tool for probing membrane mechanics and organization. This
advance opens new avenues for studying membrane-associated phenomena in
health and disease, including cancer cell mechanics, protein–membrane
interactions, and organelle dynamics.