Sharpening emitter localization in front of a tuned mirror

Single-molecule localization microscopy (SMLM) aims for maximized precision and a high signal-to-noise ratio1. Both features can be provided by placing the emitter in front of a metal-dielectric nanocoating that acts as a tunedmirror2–4. Here, we demonstrate that a higher photon yield at a lower background on biocompatible metal-dielectric nanocoatings substantially improves SMLM performance and increases the localization precision by up to a factor oftwo. The resolution improvement relies solely on easy-to-fabricate nanocoatings on standard glass coverslips and is spectrally and spatially tunable by the layer design and wavelength, as experimentally demonstrated for dual-color SMLM in cells.Publisher PDFPeer reviewe

ACCESS-OM2 v1.0: a global ocean-sea ice model at three resolutions

We introduce ACCESS-OM2, a new version of the ocean–sea ice model of the Australian Community Climate and Earth System Simulator. ACCESS-OM2 is driven by a prescribed atmosphere (JRA55-do) but has been designed to form the ocean–sea ice component of the fully coupled (atmosphere–land–ocean–sea ice) ACCESS-CM2 model. Importantly, the model is available at three different horizontal resolutions: a coarse resolution (nominally 1∘ horizontal grid spacing), an eddy-permitting resolution (nominally 0.25∘), and an eddy-rich resolution (0.1∘ with 75 vertical levels); the eddy-rich model is designed to be incorporated into the Bluelink operational ocean prediction and reanalysis system. The different resolutions have been developed simultaneously, both to allow for testing at lower resolutions and to permit comparison across resolutions. In this paper, the model is introduced and the individual components are documented. The model performance is evaluated across the three different resolutions, highlighting the relative advantages and disadvantages of running ocean–sea ice models at higher resolution. We find that higher resolution is an advantage in resolving flow through small straits, the structure of western boundary currents, and the abyssal overturning cell but that there is scope for improvements in sub-grid-scale parameterizations at the highest resolution

The state of the Martian climate

60°N was +2.0°C, relative to the 1981–2010 average value (Fig. 5.1). This marks a new high for the record. The average annual surface air temperature (SAT) anomaly for 2016 for land stations north of starting in 1900, and is a significant increase over the previous highest value of +1.2°C, which was observed in 2007, 2011, and 2015. Average global annual temperatures also showed record values in 2015 and 2016. Currently, the Arctic is warming at more than twice the rate of lower latitudes

Audiovisual: Expansion Microscopy on Saccharomyces cerevisiae

Supplementary Video 1. The video contains footage and instructions showing how to proceed throughout the protocol, focusing on the most critical steps.Related Publication

Text: Expansion Microscopy on Saccharomyces cerevisiae

Supplementary figures of custom tools and troubleshooting table.Related Publication

Automated AFM analysis of DNA bending reveals initial lesion sensing strategies of DNA glycosylases

Base excision repair is the dominant DNA repair pathway of chemical modifications such as deamination, oxidation, or alkylation of DNA bases, which endanger genome integrity due to their high mutagenic potential. Detection and excision of these base lesions is achieved by DNA glycosylases. To investigate the remarkably high efficiency in target site search and recognition by these enzymes, we applied single molecule atomic force microscopy (AFM) imaging to a range of glycosylases with structurally different target lesions. Using a novel, automated, unbiased, high-throughput analysis approach, we were able to resolve subtly different conformational states of these glycosylases during DNA lesion search. Our results lend support to a model of enhanced lesion search efficiency through initial lesion detection based on altered mechanical properties at lesions. Furthermore, its enhanced sensitivity and easy applicability also to other systems recommend our novel analysis tool for investigations of diverse, fundamental biological interactions

Septins Have a Dual Role in Controlling Mitotic Exit in Budding Yeast

AbstractIn Saccharomyces cerevisiae, the spindle position checkpoint ensures that cells do not exit mitosis until the mitotic spindle moves into the mother/bud neck and thus guarantees that each cell receives one nucleus [1–6]. Mitotic exit is controlled by the small G protein Tem1p. Tem1p and its GTPase activating protein (GAP) Bub2p/Bfa1p are located on the daughter-bound spindle pole body. The GEF Lte1p is located in the bud. This segregation helps keep Tem1p in its inactive GDP state until the spindle enters the neck. However, the checkpoint functions without Lte1p and apparently senses cytoplasmic microtubules in the mother/bud neck [7–9]. To investigate this mechanism, we examined mutants defective for septins, which compose a ring at the neck [10]. We found that the septin mutants sep7Δ and cdc10Δ are defective in the checkpoint. When movement of the spindle into the neck was delayed, mitotic exit occurred, inappropriately leaving both nuclei in the mother. In sep7Δ and cdc10Δ mutants, Lte1p is mislocalized to the mother. In sep7Δ, but not cdc10Δ, mutants, inappropriate mitotic exit depends on Lte1p. These results suggest that septins serve as a diffusion barrier for Lte1p, and that Cdc10p is needed for the septin ring to serve as a scaffold for a putative microtubule sensor