High-throughput analysis of membrane fluidity unveils a hidden dimension in immune cell states

Abstract

Cell membranes undergo biophysical remodelling as an adaptation to the surroundings and to perform specific biological functions. However, the extent and relevance of such changes in human immune systems remain unknown, largely due to the lack of high throughput and multidimensional methodologies. Here, we describe a cytometry-based method with single-cell resolution which fills this technological gap by combining biophysical profiling with conventional biomarker analysis. This platform allows to reveal notable cell type-dependent remodelling of membrane fluidity during immune stimulations and in diseases. Using immune cells exposed to tumour microenvironment as well as from long COVID and chronic lymphocyte leukaemia patients, we demonstrate that membrane fluidity is orthogonal to surface marker expression. Moreover, this biophysical parameter identifies new functional and pathological states of immune cells previously undetected via surface marker profiling alone. Our findings will contribute to a more precise definition of immune cell states based on their biophysical properties and will pave the way for a better understanding of the functional heterogeneity of immune cells.

VISION — an open-source software for automated multi-dimensional image analysis of cellular biophysics

“…For example, researchers have demonstrated how remodelling of membrane biophysical properties, including polarity, tension, fluidity, and depolarization, can influence processes such as cell proliferation and migration (Atilla-Gokcumen et al, 2014;Braig et al, 2015;Edmond et al, 2015;Kunduri, Acharya and Acharya, 2022) and immune response (Rentero et al, 2008;Rudd-Schmidt et al, 2019;Tello-Lafoz et al, 2021). Recent advances in technology for high-throughput characterization of cell biophysics, have highlighted the importance of such biophysical remodelling also in diseases (Andronico et al, 2024). Among others, environment-sensitive probes are often used to investigate alterations in cellular biophysics.…”