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HAJJ, B.

Publications and source records attributed to HAJJ, B..

2 recordsLinked to original sources

Unexpected softening of giant unilamellar vesicles by budding yeast septin filaments: a curvature dependent mechanism

Budding yeast septins assemble into filamentous networks bound to the inner plasma membrane. In situ or in vitro, septins are implicated in membrane deformations. We therefore suspected that septins might alter membrane mechanical properties both directly or indirectly. To decipher whether septins directly tune the rigidity of membranes, we used a cell free in vitro approach. To this end, using AFM, we measured the mechanical response of reconstituted GUVs pre-incubated with septins. Unexpectedly, we find that large GUVs (typically tens of {micro}m diameter size) are more deformable in the presence of septins. Theoretical modeling suggests that this peculiar behavior is likely due to initial micrometer membrane "wrinkled" deformations imposed by septins. Conversely, small GUVs (1 to 2 microns in diameter) cannot undergo any micrometric deformations and are thereby less deformable with septin filaments bound. Our findings suggest that, in specific cellular context, septins could provide a membrane reservoir and eventually facilitate membrane deformations. Significance statementFilamentous cytoskeletal septins, interacting with membranes would be expected to enhance membrane rigidity. Upon mechanical stress, GUVs larger than tens of microns appear, more deformable in the presence of septins. Septins initial membrane reshaping is responsible for this unexpected behavior, as shown by theoretical modeling. However smaller non deformable vesicles are more rigid, with septins bound.

biophysics↗

Selective volumetric excitation and imaging for single molecule localization microscopy in multicellular systems

Light sheet fluorescence microscopy (LSFM) has become a leading standard in high-resolution imaging of living samples in 2- and 3-dimensions. Biological samples are however not restricted to a single observation plane and several molecular processes evolve rapidly in 3D. The conventional mechanical scanning required in LSFM limits the range of observable dynamics and are usually restricted in resolution. Here we introduce a new strategy for instantaneous volumetric excitation and volumetric imaging of single-molecules in cell aggregates. The technique combines, for the first time, the use of light sheet microscopy and multifocus microscopy (MFM) and enables a volumetric 4D imaging of biological samples with single-molecule resolution. We engineered the excitation beam to yield a modular and uniform excitation matching the observable detection range of MFM. The strength of the method is highlighted with examples of single-molecule 3D tracking and 3D super-resolution imaging in multicellular samples.

biophysics↗