Compressive forces driven by lateral actin fibers are a key to the nuclear deformation under uniaxial cell-substrate stretching.

Cells sense the direction of mechanical stimuli including substrate stretching and show morphological and functional responses. The nuclear deformation with respect to the direction of mechanical stimuli is thought of as a vital factor in mechanosensitive intracellular signaling and gene transcription, but the detailed relationship between the direction of stimuli and nuclear deformation behavior is not fully solved yet. Here, we assessed the role of actin cytoskeletons in nuclear deformation caused by cell substrate stretching with different directions. Cells on a PDMS stretching chamber were subjected to a step-strain and changes of long- and short-axes of nucleus before and after stretching were evaluated in terms of nuclear orientation against the direction of stretching. Nuclei oriented parallel to the stretching direction showed elongation and shrinkage in the long and short axes, respectively, and vice versa. However, calculation of the aspect ratio (ratio of long- and short-axes) changes revealed orientation-depend nuclear deformation: The nucleus oriented parallel to the stretching direction showed a greater aspect ratio change than it aligned in the perpendicular direction of the stretching. A decrease in actin cytoskeletal tension significantly changed the nuclear deformation only in the short axis direction, thereby abolishing the orientation-depend deformation of the nucleus. These results suggest that lateral compressive forces exerted by the actin cytoskeleton is a key factor of orientation-depend deformation in short axis of the nucleus under the cell-substrate stretching condition, and may be crucial for mechano-sensing and responses to the cell-substrate stretching direction.

Intranuclear strain in living cells subjected to substrate stretching: A combined experimental and computational study.

Nuclear deformation caused by mechanical stimuli has been suggested to significantly impact various cellular activities, such as gene expression, protein synthesis and mechanotransduction. To understand how nuclear deformation regulates cellular behaviors, the details of intranuclear strain distribution caused by mechanical stimuli as well as intranuclear mechanical properties are required. Here, we examine local mechanical strains within the nucleus in a living cell subjected to substrate stretching and estimate the local nuclear mechanical properties. A HeLa cell in a PDMS chamber was subjected to a 10% step-strain by using a custom-made uni-axial stretching device. Local displacements and the distribution of the equivalent strain within the nucleus were obtained from fluorescence images of the nucleus before and after the application of stretching. The intranuclear strain showed heterogeneous distribution, and higher strain regions were observed not only at the center, but also periphery of the nucleus. We examined the role of the chromatin condensation level and actin cytoskeleton by treating cells with Trichostatin A and Cytochalasin D, respectively. Interestingly, these treatments did not cause significant changes in the intranuclear strain distribution. Referring to the experimental results, we reproduced the nuclear strain distribution in a finite element model to estimate relative distribution of Young's modulus within the nucleus, and observed substantially lower Young's modulus levels in the peripheral regions of the nucleus relative to those found in the central regions of the nucleus. We reveal heterogeneous strain distribution within the nucleus in a living cell subjected to substrate stretching, and the results provide insights into the importance of heterogeneity of intranuclear mechanical properties.

Effective atomic number image determination with an energy-resolving photon-counting detector using polychromatic X-ray attenuation by correcting for the beam hardening effect and detector response.

In this study, we propose an effective atomic number (Zeff) determination method based on a photon-counting technique. The proposed method can correct for the beam hardening effect and detector response based on polychromatic X-rays to allow high accuracy material identification. To demonstrate the effectiveness of our method, the procedure was applied to X-ray images acquired by a prototype energy-resolving photon-counting detector and we obtained an Zeff image with accuracy of Zeff ± 0.5 regardless of the mass thickness.