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Nidriche, A.

Publications and source records attributed to Nidriche, A..

3 recordsLinked to original sources

Polarized neutrons for the study of individual and collective fast dynamics in proteins

Neutron scattering is a powerful technique to investigate atomic structures and molecular dynamics of proteins at the nano-scale. When it comes to dynamics, incoherent and coherent scattering respectively provide information on the single and collective dynamics of nuclei. In proteins, hydrogen has the highest incoherent cross-section, and it is common practice to overlook the contribution of coherent terms stemming from all nuclei. However, the fast collective dynamics of heavier nuclei could also be studied if coherent scattering and incoherent scattering were experimentally separated. The recent advent of polarized neutron spectroscopy with sufficient flux and energy resolution has made it possible, and opens new perspectives to investigate the relative importance of coherent scattering and the information it provides on biological samples. The present study reports on the use of polarized quasi-elastic neutron scattering (QENS) and the application of a minimalistic model adapted to both individual and collective dynamics. Using a perdeuterated green fluorescent protein as a model globular protein, the study provides an interpretation of the dynamical parameters obtained with QENS, and a comparative study of the Elastic Coherent and Incoherent Scattering Factor. Based on both experiments and calculations, we discuss the relative importance of distinct and self components of coherent scattering, which is often wrongly assumed to be representative of collective dynamics only. The results highlight the current impediments rendering complicated a straightforward analysis of fast collective dynamics in hydrated protein samples.

biophysics↗

Influence of Non-Specific Surface Adhesion on the Shape and Microrheology of Red Blood Cells

Poly-L-Lysine (PLL) mediates the non-specific adhesion of cells and is commonly used in Atomic Force Microscopy (AFM) measurements, to ensure that cells remain attached to the substrate. However, it is acknowledged that adhesion affects the measured mechanical properties, in particular in the case Red Blood Cells (RBCs). This results in a wide range of Youngs modulus E reported in the literature. The present study aims at providing a systematic approach to the impact of non-specific adhesion on the rheology of RBCs. It provides a correlation between the topography profile of adherent RBCs and their rheology, from weak (cPLL = 10-3 mg/mL) to strong-adhesion (cPLL = 100 mg/mL) regimes. Using RICM and AFM, we find that there is a continuum of RBC shapes promoted by adhesion, from concave to dome-shaped, as predicted by the theory of vesicle adhesion. Their elastic properties discriminate them into two populations depending on adhesion strength, where stiffer RBCs (E {gtrsim} 100 Pa) correlate with dome-shaped cells. These findings are supported by rheology measurements of the dynamic complex shear modulus G*(f): while the storage modulus increases with cell-substrate adhesion, reflective of an increased membrane shear modulus, the loss modulus remains unchanged. Finally, further analysis inspired by membrane theory shows that different deformation modes may be triggered during indentation of either weakly or strongly adhering RBCs, illustrating the limits of the Hertz model.

biophysics↗

Quantifying optical sectioning in reflection microscopy with patterned illumination

Sectioning techniques based on patterned illumination have been widely used to obtain well-contrasted images of thick samples using widefield imaging setups. While their application to fluorescence microscopy has been extensively demonstrated and studied, their application to reflection imaging is scarcer and their performance has only been partly characterized. In this paper, we study numerically and analytically two such sectioning techniques, line confocal (LC) and structured illumination microscopy (SIM), in the context of their application to coherent reflection imaging. We derive approximate analytical equations to relate the performance of sectioning to the optical setup parameters, allowing straightforward understanding of their influence on the image intensity and depth of focus, and we systematically compare our predictions with experimental data. Finally, we quantify the precision and accuracy of each method in typical practical cases, providing guidelines to choose the most appropriate (LC, SIM, or a simple background subtraction on a widefield image) for the sample under study. We illustrate optical sectioning in the particular case of reflection interference contrast (RIC) microscopy, an imaging technique widely used in soft matter and biophysics studies to monitor object-surface interactions, or quantify surface functionalization. Our derivation, however, should also prove useful for other reflection methods such as optical coherence tomography (OCT) or flood illumination ophtalmoscopy.

biophysics↗