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Rufin, M.

Publications and source records attributed to Rufin, M..

2 recordsLinked to original sources

Centrosome Softening As A Mechanical Adaptation For Mitosis

Centrosomes are microtubule-organizing centers important for mitotic spindle assembly and chromosome segregation. During mitosis, centrosomes are exposed to mechanical forces via the microtubules they nucleate, yet the material properties underlying their response to these forces remain poorly understood. In this study, we systematically probed the mechanical behavior of C. elegans centrosomes, both in vitro and in vivo. Using microtubule perturbations and quantitative live cell imaging, we found that centrosomes become increasingly deformed during mitosis. Centrosome deformation is independent of cortical pulling forces but instead results from microtubule polymerization within the pericentriolar material. This deformation impacts centrosome size: as microtubule number decreases with cell volume in early cleavage divisions, centrosome size scales proportionately. To directly measure centrosome elasticity, we employed atomic force microscopy (AFM) on isolated centrosomes in vitro and Brillouin light scattering microscopy in developing embryos in vivo. Both approaches revealed that centrosomes progressively soften during mitosis. Theoretical modeling predicts that softening serves to dampen spindle force fluctuations, helping to protect kinetochore-microtubule interactions and safeguarding chromosome segregation. Further, softening may enhance centrosomal microtubule nucleation capacity, facilitating mitotic spindle assembly, particularly in large early embryonic cells. We propose that centrosome softening is a mechanical adaptation for mitosis that couples microtubule number to centrosome size through force-dependent deformation. This optimally balances two mitotic requirements: the need for robust microtubule nucleation and the ability to withstand spindle forces, thereby ensuring accurate cell division.

cell biology↗

Refractive index mapping below the diffraction limit via single molecule localization microscopy

Single molecule localization microscopy (SMLM) is a powerful method to image biological samples in three dimensions below the diffraction limit of light microscopy. Beyond the position of the emitter, the shape of the single molecule point spread function provides additional information, for example about the refractive properties of the sample between the emitter and the glass coverslip. Here, we show that combination of SMLM with atomic force microscopy (AFM) allows to map the refractive index of a biological sample at sub-diffraction resolution and at a precision only limited by measurement errors of SMLM and AFM. We showcase the new method by the determination of the refractive index of isolated single collagen fibrils. Variabilities both in refractive index and the swelling behavior of single fibrils upon drying and rehydration exposed deviations from the ensemble behavior, demonstrating differential hydration of single collagen fibrils. Mapping the refractive index along single collagen fibrils revealed substantial fluctuations at characteristic length scales below 500 nm, which indicates structural heterogeneity of collagen fibrils at the length scale of single collagen molecules.

biophysics↗