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Purdie et al. use label-free fluorescence lifetime imaging microscopy to measure metabolic changes in human liver biopsies and experimental models of steatotic liver disease. This approach directly reports on metabolic dysfunction consistently across samples, substantiated by extensive analysis of structural and functional mitochondrial dysfunction.

In vitro models of actin organization show the formation of vortices, asters and stars. Here Fritzsche et al. show that such actin structures form in living cells in a manner dependent on the Arp2/3 complex but not myosin, and such structures influence membrane architecture but not cortex elasticity.

Much of our understanding of the role of actin in cell migration is based on studies of cells moving across two-dimensional surfaces. Wilson et al.show that cells crawling in three dimensions through a narrow channel form two functionally distinct actin networks at the leading edge.

Current preclinical models fail to capture the mechanics of oligodendrocyte myelination. Lasli et al.¹ now demonstrate that the mechanical compliance of the axonal niche is a key determinant of oligodendrocyte maturation. By developing a platform that mimics the extreme mechanical softness of the central nervous system, they reveal myelination as a mechanically gated process as much as a biochemically regulated one.

Methods to assess diffusion dynamics and oligomerisation of biomolecules remain scarce. Here, the authors present a brightness-transit statistics method to measure diffusion and oligomerisation simultaneously with high sensitivity, and they apply their approach to study the organisation of CD40.

Quantifying rapidly progressing three-dimensional forces generated by cells remains a major challenge in mechanobiology. Here, the authors show that combining traction force microscopy with astigmatic imaging permits sensitive out-of-plane force estimation on the second timescale.

Quantifying rapid and small cellular forces is a major challenge in mechanobiology. Here, the authors show a >2-fold spatially and >10-fold temporally force sampling improvement combining traction force microscopy with total internal reflection fluorescence super-resolution structured illumination microscopy.

Interactions between Semaphorin-3A (SEMA3A) and Neuropilin-1 (NRP1) and Plexin-A1 and Plexin-A4 have been shown to affect T cell development. Here the authors investigate how these interactions affect CD8⁺ T cells in tumour immunity, showing that NRP-1, Plexin-A1 and Plexin-A4 are upregulated on T cells allowing tumour derived SEMA3A to inhibit CD8⁺ T cell migration and function.

Analogue regulation of gene expression is important for normal function in mammals but existing genetic technologies are designed to achieve ON/OFF control. Here the authors develop synthetic microRNA silencing-mediated fine-tuners (miSFITs) to precisely control target gene expression levels.

T cells communicate with antigen-presenting cells (APC) via the signaling crosstalk at the immunological synapse (IS). Here the authors use bead-supported lipid bilayers as synthetic APCs to find that trans-synaptic vesicles produced by T cells in the IS carry specialized cargos distinct from constitutive extracellular vesicles to serve as intercellular messengers.

The early embryo maintains its structure in the face of large mechanical stresses during morphogenesis. Here they show that ASPP2 acts to preserve epithelial integrity in regions of high apical tension during early development.

New anti-fibrotics aided by a better understanding of fibrotic cell subsets and their functions are needed. Here the authors perform single-cell RNA-seq and CyTOF on nodules from patients with Dupuytren’s disease to map subsets and the state of mesenchymal cells during pathogenic fibrogenesis.

It has been suggested that the cytoplasm of living cells can be described as a porous elastic meshwork bathed in an interstitial fluid. Microindentation tests now show that intracellular water redistribution plays a fundamental role in cellular rheology and that at physiologically relevant timescales cellular responses to mechanical stresses are consistent with such a poroelastic model.

Mechanical forces play important roles in cell biology and traction force microscopy (TFM) experiments have enabled quantification of the cell-generated forces when placed on substrates of distinct stiffnesses. Here the authors evaluate the effect of the Poisson’s ratio- one of the main descriptors of the material’s mechanical behaviour together with the Elastic Modulus, in the context of TFM experiments.

Huw Colin-York et al. use advanced microscopy techniques to show that the cortical actin network within a model mast cell line undergoes a series of reorganizational events at the basal interface during activation. They find that actin patterns co-localize with zones of Arp2/3 nucleation and myosin-II activity accompanies network reassembly.

Many cellular processes rely on cells generating or responding to nanoscale mechanical forces. This protocol describes STED–traction force microscopy (STFM), which allows these forces to be measured with higher resolution and accuracy than standard TFM.

This protocol uses fluorescence recovery after photobleaching (FRAP) to dissect the reaction dynamics leading to protein turnover in macromolecular complexes in living cells.

Communications Biology is inviting submissions on the topic of live microscopy – from new tools to emerging techniques, from conventional to advanced light microscopy - with the aim of publishing high-quality research devoted to advance our understanding of biology.

We are inviting submissions of articles on the role of mechanobiology in health and disease with the aim of publishing high quality research devoted to advance our understanding of mechanics shaping biological function. We are also happy to present a Collection of papers already published in our journal in this exciting field.