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Brightfield microscopy, as expected, lacked sufficient contrast to discern cellular details and cellular boundaries were not readily identified.
Defined signals that dictate the architecture of cellular boundaries in confluent cultures are poorly characterized.
On the other hand, cellular boundaries of microcolonies were distinct on all substrates for S. aureus microcolonies (cf. Figs 2c, 3c and 4c).
The interfacial energy mismatch between hydrophilic EPS and hydrophobic substrates can account for the disappearance of cellular boundaries for E. coli O157 H7 microcolonies.
The physical properties of membrane interfaces, quasi-two dimensional, bilayer lipid membranes, which together with membrane proteins delimit cellular boundaries, are now largely well-understood at thermodynamic equilibrium.
We measured embryos at stages 14 and 15, following the first point (stage 13)17,20 at which the pallium and subpallium can be distinguished by cellular boundaries (for example, the PSB).
Similar(18)
For BeWo cells, sodium fluorescein transport was reduced to 31.49 ± 3.07%, 25.29 ± 3.55% and 18.90 ± 2.01% at day 3, 5 and 7 post seeding indicating a tight cellular boundary even at early stages of this in vitro barrier model.
Regarding the formation of microcolonies without any visible cellular boundary on hydrophobic substrates but not on hydrophilic ones for E. coli O157 H7 (cf. Figs 2f, 3f and 4f), we consider the components of EPS and its wetting characteristics.
The fact that amphiphilic compounds present in meteorites can self-assemble into bilayer membranes makes it plausible that lipid-like molecules were available on the early Earth and could provide the first cellular boundary structures.
In contrast, engrailed might play a different role in Onychophora, as the early expression domain of this gene extends beyond the segmental furrow and no precise cellular boundary is evident between engrailed and wingless domains in the onychophoran embryo [ 41].
In the 10× images, the cellular boundary was estimated from fluorescence levels in the α-tubulin channel by extension from nuclei by the propagation algorithm (Jones et al., 2005).
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