Abstract
Cellular function is reliant on the dynamic interplay between the plasma membrane and the actin cytoskeleton. This critical relationship is of particular importance in immune cells, where both the cytoskeleton and the plasma membrane work in concert to organize and potentiate immune signaling events. Despite their importance, there remains a critical gap in understanding how these respective dynamics are coupled, and how this coupling in turn may influence immune cell function from the bottom up. In this review, we highlight recent optical technologies that could provide strategies to investigate the simultaneous dynamics of both the cytoskeleton and membrane as well as their interplay, focusing on current and future applications in immune cells. We provide a guide of the spatio-temporal scale of each technique as well as highlighting novel probes and labels that have the potential to provide insights into membrane and cytoskeletal dynamics. The quantitative biophysical tools presented here provide a new and exciting route to uncover the relationship between plasma membrane and cytoskeletal dynamics that underlies immune cell function.
Highlights
It is becoming increasingly clear that the dynamics of the cortical actin cytoskeleton and the plasma membrane are intimately linked to immune cell function, playing a critical role in, for instance, the regulation of receptor organization, granule section, and specific cytoskeletal protrusions [1,2,3,4]
Both total internal reflection fluorescence (TIRF) and singleplane illumination (SPIM) schemes have been combined with camera based fluorescence correlation spectroscopy (FCS) acquisitions yielding similar and even larger statistics compared to scanning fluorescence correlation spectroscopy (sFCS) [91,92,93]
The communication of immune cells with their environment, other immune cells and target cells involves a diversity of complex receptor-ligand interactions
Summary
It is becoming increasingly clear that the dynamics of the cortical actin cytoskeleton and the plasma membrane are intimately linked to immune cell function, playing a critical role in, for instance, the regulation of receptor organization, granule section, and specific cytoskeletal protrusions [1,2,3,4]. Little is known about how these interactions influence the behavior of cells in more complex tissue environments or in their full physiological setting, with our knowledge often restricted to in vitro single cell studies (Figure 1C) This has primarily been due to a lack of accessible technologies with sufficient temporal (< ms binding and transport events) and spatial resolution (single proteins on the order of nanometres can effect changes on cellular level beyond tenth of micrometers) to assess the correlated dynamics of the plasma membrane and the actin cytoskeleton without perturbing the system.
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