Spicule movement on RBCs during echinocyte formation and possible segregation in the RBC membrane.

We use phase contrast microscopy of red blood cells to observe the transition between the initial discocyte shape and a spiculated echinocyte form. During the early stages of this change, spicules can move across the surface of the cell; individual spicules can also split apart into pairs. One possible explanation of this behaviour is that the membrane forms large scale domains in association with the spicules. The spicules are formed initially at the rim of the cell and then move at speeds of up to 3 µm/min towards the centre of the disc. Spicule formation that was reversed and then allowed to proceed a second time resulted in spicules at reproducible places, a shape memory effect that implies that the cytoskeleton contributes towards stopping the spicule movement. The splitting of the spicules produces a well-defined shape change with an increase in membrane curvature associated with formation of the daughter pair of spicules; the total boundary length around the spicules also increases. Following the model in which the spicules are associated with lipid domains, these observations suggest an experimental procedure that could potentially be applied to the calculation of the line tension of lipid domains in living cells.

2D protein arrays induce 3D in vivo-like assemblies of cells.

We report on the ability of two-dimensional protein crystals to induce the formation of homo- and heterotypic multicellular spheroids (MCSs) which resemble the morphology and hierarchical organization of living tissues and tumours. We have systematically studied the influence of the initial cell density and incubation time on the kinetics of spheroid growth and spheroid lifespan. Hereby a novel methodology has been established to produce MCSs on protein-based molecular layers.

Ultra-fast laser microprocessing of medical polymers for cell engineering applications.

Picosecond laser micromachining technology (PLM) has been employed as a tool for the fabrication of 3D structured substrates. These substrates have been used as supports in the in vitro study of the effect of substrate topography on cell behavior. Different micropatterns were PLM-generated on polystyrene (PS) and poly-L-lactide (PLLA) and employed to study cellular proliferation and morphology of breast cancer cells. The laser-induced microstructures included parallel lines of comparable width to that of a single cell (which in this case is roughly 20µm), and the fabrication of square-like compartments of a much larger area than a single cell (250,000µm(2)). The results obtained from this in vitro study showed that though the laser treatment altered substrate roughness, it did not noticeably affect the adhesion and proliferation of the breast cancer cells. However, pattern direction directly affected cell proliferation, leading to a guided growth of cell clusters along the pattern direction. When cultured in square-like compartments, cells remained confined inside these for eleven incubation days. According to these results, laser micromachining with ultra-short laser pulses is a suitable method to directly modify the cell microenvironment in order to induce a predefined cellular behavior and to study the effect of the physical microenvironment on cell proliferation.

Capacitive scanning dilatometry and frequency-dependent thermal expansion of polymer films

The dilatometric properties of polymer films near and above their glass-transition temperatures were explored using capacitive high-frequency detection in temperature ramping as well as in harmonic temperature cycling experiments. The broad applicability of capacitive scanning dilatometry is demonstrated by the investigation of macromolecular systems of vastly different polarity such as polystyrene, polybutadiene, and polyvinylacetate. From temperature cycling experiments the real and imaginary parts of the frequency-dependent thermal-expansion coefficient are determined in the sub-Hz regime.