Forces play a crucial role in biological mechanisms, such as cellular response, molecular interactions, and protein binding, and are essential for deriving the nanomechanical properties of a sample. AFM has emerged as a key platform for the precise measurement of interaction forces on the nanometer scale. AFM force spectroscopy is used to quantify forces and determine nanomechanical properties such as Young's modulus, cell adhesion, and viscoelastic properties, information that is invaluable for studying interaction-based and disease-related biomechanical changes. Prof. Janshoff will outline his work using AFM to investigate the biomechanical properties of cell membranes, and, in particular, the viscoelastic behavior of the cell cortex.
Webinar Highlights:
The mechanical properties of cells are intricately linked to the architecture and dynamics of their viscoelastic cortex. This cortex comprises a contractile, cross-linked actin mesh that attaches to the plasma membrane via linker proteins. However, our understanding of cell mechanics has primarily focused on the upper, apical side of adherent, polarized epithelial cells due to their accessibility in culture dishes. Consequently, less is known about the viscoelastic properties of basal membranes. In our investigation, we explored the viscoelastic behavior of basolateral membranes derived from polarized MDCK II epithelial cells. These membranes were subjected to external deformation, and their response was compared to that of living cells probed at the apical side. To achieve this, MDCK II cells were cultured on porous surfaces until they formed a confluent layer. The upper cell body was then removed using a squirting-lysis protocol, leaving behind free-standing basal membranes. Through force indentation and relaxation experiments, I precisely assessed the cortical viscoelasticity of these membranes. Interestingly, when comparing the data to reconstituted neat lipid bilayers using giant liposomes and porespanning membranes in conjunction with atomic force microscopy, a significant difference emerged: basal membranes exhibited an absence of energy dissipation. However, we were able to rescue this behavior by artificially attaching an actin cortex to the bilayer. Furthermore, I developed a theoretical framework to describe force cycles and applied it to obtain the timedependent area compressibility modulus of biomembranes. When compared to the viscoelastic response of living cells, biomembranes derived from living cells were found to be substantially less fluid and stiffer. Nevertheless, they still adhered to the same universal scaling law, provided excess area was correctly accounted for. This research sheds light on the fascinating biomechanical properties of cell membranes and their implications in cell biology.
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Andreas Janshoff, Professor of Biophysical Chemistry, University of Goettingen