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Biology subjects

Suarez, I. A. R.

Publications and source records attributed to Suarez, I. A. R..

2 recordsLinked to original sources

Nuclear morphology and chromatin organization modulate T cell cytoskeletal remodeling and immune synapse formation

T cell activation is characterized by rapid reorganization of the actin cytoskeleton and cell spreading on the antigen presenting cell. The T cell nucleus occupies a large fraction of the cell volume, and its mechanical properties are likely to act as a key determinant of activation. However, the contribution of nuclear mechanics to T cell spreading and activation is not well understood. Mechanical rigidity of lymphocyte nuclei is conferred by chromatin compaction and dense packing of heterochromatin. We find that nuclear deformation and increased chromatin compaction accompany T cell spreading, in T cells. Reducing chromatin compaction leads to increased cell spread area and nuclear deformation, while diminishing accumulation and peripheral enrichment of F-actin at the immune synapse. In contrast, enhanced chromatin compaction reduced spread area and nuclear deformation, which was accompanied by increased peripheral F-actin organization at the immune synapse. These findings suggest a reciprocal interaction between chromatin compaction and actin cytoskeletal organization. We identified SUN proteins and myosin as critical elements through which chromatin compaction orchestrates actin morphology and cell shape, facilitating T cell adaptation to antigen-presenting surfaces of varying stiffness. These results emphasize the crucial role of chromatin compaction in T cell activation, underlining the mechanical relationship between the nucleus and the cytoskeleton during immune responses, and suggest new avenues for understanding T cell mechano-responsiveness.

biophysics↗

Three-dimensional structured illumination microscopy with enhanced axial resolution

We present two distinct, complementary methods for improving axial resolution in three-dimensional structured illumination microscopy (3D SIM) with minimal or no modification to the optical system. First, we show that placing a mirror directly opposite the sample enables 4-beam interference with higher spatial frequency content than 3D SIM illumination, offering near-isotropic imaging with [~]120 nm lateral and 160 nm axial resolution. Second, we develop an improved deep learning method that can be directly applied to 3D SIM data, obviating the need for additional hardware. This procedure results in [~]120 nm isotropic resolution and can be combined with denoising to facilitate volumetric imaging spanning dozens of time points. We demonstrate the potential of these advances by imaging a variety of cellular samples, delineating the nanoscale distribution of vimentin and microtubule filaments, observing the relative positions of caveolar coat proteins and lysosomal markers, and visualizing rich cytoskeletal dynamics within T-cells in the early stages of immune synapse formation.

cell biology↗