![]() Moreover, microfluidics can be used to create time-varying environments. ![]() One can take advantage of microfluidic microchemostat that, unlike liquid bulk culture, enables long-term observations of cells growing as a monolayer. This is a decisive advantage to investigate a number of important biological problems, including chronological ageing, epigenetic heritability and dynamic features such as the cell cycle and circadian oscillations in non-synchronized cell populations. The former provides great statistical details on the diversity of the studied cell population, whereas the latter provides longitudinal information on single cells: individual cells can be tracked in time. Used in combination with fluorescence reporter techniques, flow cytometry and time-lapse microscopy are arguably the two most widely employed quantitative single-cell observation approaches. Observing cellular processes at the single cell level is often necessary to understand how cells respond to endogenous and environmental changes. As a community effort, we set up a website, the Yeast Image Toolkit, with the benchmark and the Evaluation Platform to gather this and additional information provided by others. We created a benchmark dataset with manually analysed images and compared CellStar with six other tools, showing its high performance, notably in long-term tracking. A graphical user interface enables manual corrections of S&T errors and their use for the automated correction of other, related errors and for parameter learning. ![]() The key features are the use of a new variant of parametrized active rays for segmentation, a neighbourhood-preserving criterion for tracking, and the use of an iterative approach that incrementally improves S&T quality. Here, we present CellStar, a tool chain designed to achieve good performance in long-term experiments. Surprisingly, even for yeast cells that have relatively regular shapes, no solution has been proposed that reaches the high quality required for long-term experiments for segmentation and tracking (S&T) based on brightfield images. This report sets the stage for classes of thermoresponsive materials to respond to macromolecule concentration rather than temperature changes.With the continuous expansion of single cell biology, the observation of the behaviour of individual cells over extended durations and with high accuracy has become a problem of central importance. We find that the transformations observed upon heating nanoemulsions above their surfactant’s LCST can instead be induced at physiological temperatures through the addition of polymers and protein, rendering thermoresponsive materials “crowding responsive.” We demonstrate that the cytosol is a stimulus for nanoemulsions, with droplet fusion occurring upon injection into cells of living zebrafish embryos. These amphiphiles stabilize oil-in-water nanoemulsions at temperatures below the LCST but are ineffective surfactants above the LCST, resulting in emulsion fusion. We prepare poly(2-oxazoline) amphiphiles that exhibit lower critical solution temperatures (LCST) in serum above physiological temperature. We introduce a new endogenous stimulus, biomacromolecule crowding, which we achieve by leveraging changes in thermoresponsive properties of polymers upon high concentrations of crowding agents. Stimuli-responsive materials are exploited in biological, materials, and sensing applications. ![]()
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