Low Surface Potential with Glycoconjugates Determines Insect Cell Adhesion at Room Temperature

Cell-coupled field-effect transistor (FET) biosensors have attracted considerable attention because of their high sensitivity to biomolecules. The use of insect cells (Sf21) as a core sensor element is advantageous due to their stable adhesion to sensors at room temperature. Although visualization of the insect cell-substrate interface leads to logical amplification of signals, the spatiotemporal processes at the interfaces have not yet been elucidated. We quantitatively monitored the adhesion dynamics of Sf21 using interference reflection microscopy (IRM). Specific adhesion signatures with ring-like patches along the cellular periphery were detected. A combination of zeta potential measurements and lectin staining identified specific glycoconjugates with low electrostatic potentials. The ring-like structures were disrupted after cholesterol depletion, suggesting a raft domain along the cell periphery. Our results indicate dynamic and asymmetric cell adhesion is due to low electrostatic repulsion with fluidic sugar rafts. We envision the logical design of cell-sensor interfaces with an electrical model that accounts for actual adhesion interfaces.Matsuzaki T., Terutsuki D., Sato S., et al. Low Surface Potential with Glycoconjugates Determines Insect Cell Adhesion at Room Temperature. Journal of Physical Chemistry Letters 2022 13(40), 9494-9500. DOI: 10.1021/acs.jpclett.2c01673. Copyright © 2022 American Chemical Society

Mechanical guidance of self-condensation patterns of differentiating progeny

Spatially controlled self-organization represents a major challenge for organoid engineering. We have developed a mechanically patterned hydrogel for controlling self-condensation process to generate multi-cellular organoids. We first found that local stiffening with intrinsic mechanical gradient (IG > 0.008) induced single condensates of mesenchymal myoblasts, whereas the local softening led to stochastic aggregation. Besides, we revealed the cellular mechanism of two-step self-condensation: (1) cellular adhesion and migration at the mechanical boundary and (2) cell-cell contraction driven by intercellular actin-myosin networks. Finally, human pluripotent stem cell-derived hepatic progenitors with mesenchymal/endothelial cells (i.e., liver bud organoids) experienced collective migration toward locally stiffened regions generating condensates of the concave to spherical shapes. The underlying mechanism can be explained by force competition of cell-cell and cell-hydrogel biomechanical interactions between stiff and soft regions. These insights will facilitate the rational design of culture substrates inducing symmetry breaking in self-condensation of differentiating progeny toward future organoid engineering.Matsuzaki T., Shimokawa Y., Koike H., et al. Mechanical guidance of self-condensation patterns of differentiating progeny. iScience 25, 105109 (2022); https://doi.org/10.1016/j.isci.2022.105109

Preparation of mechanically patterned hydrogels for controlling the self-condensation of cells

Synthetic protocols providing mechanical patterns to culture substrate are essential to control the self-condensation of cells for organoid engineering. Here, we present a protocol for preparing hydrogels with mechanical patterns. We describe steps for hydrogel synthesis, mechanical evaluation of the substrate, and time-lapse imaging of cell self-organization. This protocol will facilitate the rational design of culture substrates with mechanical patterns for the engineering of various functional organoids. For complete details on the use and execution of this protocol, please refer to Takebe et al. (2015) and Matsuzaki et al. (2014, 2022).Matsuzaki T., Kawano Y., Horikiri M., et al. Preparation of mechanically patterned hydrogels for controlling the self-condensation of cells. STAR Protocols 4, 102471 (2023); https://doi.org/10.1016/j.xpro.2023.102471

Excitation Wavelength Dependent Three-Wave Mixing at a CO-Covered Platinum Electrode

The interfacial electronic structure of a CO-covered polycrystalline platinum electrode has been studied by using the optical second harmonic generation (SHG) and sum frequency generation (SFG) techniques with various excitation wavelengths. Although the nonlinear optical (NLO) signal was enhanced by the absorption of CO at all excitation wavelengths employed in this study, the potential dependent behaviors of the NLO signal were different between the near-infrared excitation and the visible excitation. The difference was attributed to the different mechanisms for the enhancement of the NLO signal. While the electron transition from the Fermi level of Pt to the unoccupied 2πa* orbital of adsorbed CO was considered to be coupled with the NLO photon in the case of visible excitation, the dc field induced SHG regime was applied to the result obtained by the near-infrared excitation

In Situ Optical Second Harmonic Rotational Anisotropy Measurements of an Au(111) Electrode during Electrochemical Deposition of Tellurium

The electrodeposition of Te on a single-crystalline Au(111) electrode was studied with 1064-nm-excited SH rotational anisotropy measurements. The SH rotational anisotropy was significantly changed with the first underpotential deposition (upd) of Te, and the bulk Te deposition attenuated the anisotropic character of the overall surface symmetry. The change in the SH rotational anisotropy during the first upd of Te was examined using two different models. The first model considered only the contribution of the Au(111) surface to the SHG response, while the second one took into account the contributions of both the Au(111) substrate and the adsorbed Te layer. In the former case, the change in the SH rotational anisotropy can be explained by considering the quenching of the nonlinear susceptibility of the Au(111) surface. The analysis based on the latter model resulted in a rotation angle of 608 for the adsorbed Te layer against the the Au(111) lattice. This value was not consistent with that expected from the adsorbate structure of the first upd layer of Te, i.e., (√3 x √3)R30°. Thus, the former model seems to be more appropriate to explain the present results. The SH rotational anisotropy measurement also suggests that the morphology became more isotropic after the bulk Te deposition