RESUMEN
Fluorescent proteins (FP) are frequently used for studying proteins inside cells. In advanced fluorescence microscopy, FPs can report on additional intracellular variables. One variable is the local density near FPs, which can be useful in studying densities within cellular bio-condensates. Here, we show that a reduction in fluorescence lifetimes of common monomeric FPs reports increased levels of local densities. We demonstrate the use of this fluorescence-based variable to report the distribution of local densities within heterochromatin protein 1α (HP1α) in mouse embryonic stem cells (ESCs), before and after early differentiation. We find that local densities within HP1α condensates in pluripotent ESCs are heterogeneous and cannot be explained by a single liquid phase. Early differentiation, however, induces a change towards a more homogeneous distribution of local densities, which can be explained as a liquid-like phase. In conclusion, we provide a fluorescence-based method to report increased local densities and apply it to distinguish between homogeneous and heterogeneous local densities within bio-condensates.
Asunto(s)
Núcleo Celular , Células Madre Embrionarias , Animales , Ratones , Núcleo Celular/metabolismo , Diferenciación Celular , Células Madre Embrionarias/metabolismo , Heterocromatina/metabolismo , Homólogo de la Proteína Chromobox 5RESUMEN
Ionic liquids are solvent-free electrolytes, some of which possess an intriguing self-assembly at finite length scale due to Coulombic interactions. Using a continuum framework (based on Onsager's relations), it is shown that bulk nanostructures arise via linear (supercritical) and nonlinear (subcritical) bifurcations (morphological phase transitions), which also directly affect the electrical double layer structure. A Ginzburg-Landau amplitude equation is derived and the bifurcation type is related to model parameters, such as temperature, potential, and interactions. Specifically, the nonlinear bifurcation occurs for geometrically dissimilar ions and, surprisingly, is induced by perturbations on the order of thermal fluctuations. Finally, qualitative insights and comparisons to the experimentally decaying charge layers within the electrical double layer are discussed.