Thermally reconfigurable metalens.

Reconfigurable metalenses are compact optical components composed by arrays of meta-atoms that offer unique opportunities for advanced optical systems, from microscopy to augmented reality platforms. Although poorly explored in the context of reconfigurable metalenses, thermo-optical effects in resonant silicon nanoresonators have recently emerged as a viable strategy to realize tunable meta-atoms. In this work, we report the proof-of-concept design of an ultrathin (300 nm thick) and thermo-optically reconfigurable silicon metalens operating at a fixed, visible wavelength (632 nm). Importantly, we demonstrate continuous, linear modulation of the focal-length up to 21% (from 165 µm at 20 °C to 135 µm at 260 °C). Operating under right-circularly polarized light, our metalens exhibits an average conversion efficiency of 26%, close to mechanically modulated devices, and has a diffraction-limited performance. Overall, we envision that, combined with machine-learning algorithms for further optimization of the meta-atoms, thermally reconfigurable metalenses with improved performance will be possible. Also, the generality of this approach could offer inspiration for the realization of active metasurfaces with other emerging materials within field of thermo-nanophotonics.

Publisher Correction: Super-resolution fight club: assessment of 2D and 3D single-molecule localization microscopy software.

In the version of this paper originally published, Figure 4a contained errors that were introduced during typesetting. The bottom 11° ThunderSTORM image is an xz view but was incorrectly labeled as xy, and the low x-axis value in the four line profiles was incorrectly set as -60 instead of -50. These errors have been corrected in the PDF and HTML versions of the paper.

Multi-phosphorylation reaction and clustering tune Pom1 gradient mid-cell levels according to cell size.

Protein concentration gradients pattern developing organisms and single cells. In Schizosaccharomyces pombe rod-shaped cells, Pom1 kinase forms gradients with maxima at cell poles. Pom1 controls the timing of mitotic entry by inhibiting Cdr2, which forms stable membrane-associated nodes at mid-cell. Pom1 gradients rely on membrane association regulated by a phosphorylation-dephosphorylation cycle and lateral diffusion modulated by clustering. Using quantitative PALM imaging, we find individual Pom1 molecules bind the membrane too transiently to diffuse from pole to mid-cell. Instead, we propose they exchange within longer lived clusters forming the functional gradient unit. An allelic series blocking auto-phosphorylation shows that multi-phosphorylation shapes and buffers the gradient to control mid-cell levels, which represent the critical Cdr2-regulating pool. TIRF imaging of this cortical pool demonstrates more Pom1 overlaps with Cdr2 in short than long cells, consistent with Pom1 inhibition of Cdr2 decreasing with cell growth. Thus, the gradients modulate Pom1 mid-cell levels according to cell size.

Super-resolution fight club: assessment of 2D and 3D single-molecule localization microscopy software.

With the widespread uptake of two-dimensional (2D) and three-dimensional (3D) single-molecule localization microscopy (SMLM), a large set of different data analysis packages have been developed to generate super-resolution images. In a large community effort, we designed a competition to extensively characterize and rank the performance of 2D and 3D SMLM software packages. We generated realistic simulated datasets for popular imaging modalities-2D, astigmatic 3D, biplane 3D and double-helix 3D-and evaluated 36 participant packages against these data. This provides the first broad assessment of 3D SMLM software and provides a holistic view of how the latest 2D and 3D SMLM packages perform in realistic conditions. This resource allows researchers to identify optimal analytical software for their experiments, allows 3D SMLM software developers to benchmark new software against the current state of the art, and provides insight into the current limits of the field.

Waveguide-PAINT offers an open platform for large field-of-view super-resolution imaging.

Super-resolution microscopies based on the localization of single molecules have been widely adopted due to their demonstrated performance and their accessibility resulting from open software and simple hardware. The PAINT method for localization microscopy offers improved resolution over photoswitching methods, since it is less prone to sparse sampling of structures and provides higher localization precision. Here, we show that waveguides enable increased throughput and data quality for PAINT, by generating a highly uniform ~100 × 2000 µm2 area evanescent field for TIRF illumination. To achieve this, we designed and fabricated waveguides optimized for efficient light coupling and propagation, incorporating a carefully engineered input facet and taper. We also developed a stable, low-cost microscope and 3D-printable waveguide chip holder for easy alignment and imaging. We demonstrate the capabilities of our open platform by using DNA-PAINT to image multiple whole cells or hundreds of origami structures in a single field of view.

Constriction Rate Modulation Can Drive Cell Size Control and Homeostasis in C. crescentus.

Rod-shaped bacteria typically grow first via sporadic and dispersed elongation along their lateral walls and then via a combination of zonal elongation and constriction at the division site to form the poles of daughter cells. Although constriction comprises up to half of the cell cycle, its impact on cell size control and homeostasis has rarely been considered. To reveal the roles of cell elongation and constriction in bacterial size regulation during cell division, we captured the shape dynamics of Caulobacter crescentus with time-lapse structured illumination microscopy and used molecular markers as cell-cycle landmarks. We perturbed the constriction rate using a hyperconstriction mutant or fosfomycin ([(2R,3S)-3-methyloxiran-2-yl]phosphonic acid) inhibition. We report that the constriction rate contributes to both size control and homeostasis, by determining elongation during constriction and by compensating for variation in pre-constriction elongation on a single-cell basis.