Dynamin plays an important role in clathrin-mediated endocytosis (CME) by cutting the neck of nascent vesicles from the cell membrane. Here through using gold nanorods as cargos to image dynamin action during live CME, we show that near the peak of dynamin accumulation, the cargo-containing vesicles always exhibit abrupt, right-handed rotations that finish in a short time (~0.28 s). The large and quick twist, herein named the super twist, is the result of the coordinated dynamin helix action upon GTP hydrolysis. After the super twist, the rotational freedom of the vesicle drastically increases, accompanied with simultaneous or delayed translational movement, indicating that it detaches from the cell membrane. These observations suggest that dynamin-mediated scission involves a large torque generated by coordinated actions of multiple dynamins in the helix, which is the main driving force for vesicle scission.
The insightful comprehension of in situ catalytic dynamics at individual structural defects of two-dimensional (2D) layered material, which is crucial for the design of high-performance catalysts via defect engineering, is still missing. Here, we resolved single-molecule trajectories resulted from photocatalytic activities at individual structural features (i.e., basal plane, edge, wrinkle, and vacancy) in 2D layered indium selenide (InSe) in situ to quantitatively reveal heterogeneous photocatalytic dynamics and surface diffusion behaviors. The highest catalytic activity was found at vacancy in a four-layer InSe, up to ~30× higher than that on the basal plane. Moreover, lower adsorption strength of reactant and slower dissociation/diffusion rates of product were found at more photocatalytic active defects. These distinct dynamic properties are determined by lattice structures/electronic energy levels of defects and layer thickness of supported InSe. Our findings shed light on the fundamental understanding of photocatalysis at defects and guide the rational defect engineering.
Gold nanoparticles (AuNPs) have become increasingly useful in recent years for their roles in nanomedicine, cellular biology, energy storage and conversion, photocatalysis, and more. At the single-particle level, AuNPs have heterogeneous physical and chemical properties which are not resolvable in ensemble measurements. In the present study, we developed an ultrahigh-throughput spectroscopy and microscopy imaging system for characterization of AuNPs at the single-particle level using phasor analysis. The developed method enables quantification of spectra and spatial information on large numbers of AuNPs with a single snapshot of an image (1024 × 1024 pixels) at high temporal resolution (26 fps) and localization precision (sub-5 nm). We characterized the localized surface plasmonic resonance (SPR) scattering spectra of gold nanospheres (AuNSs) of four different sizes (40–100 nm). Comparing to the conventional optical grating method which suffers low efficiency in characterization due to spectral interference caused by nearby nanoparticles, the phasor approach enables high-throughput analysis of single-particle SPR properties in high particle density. Up to 10-fold greater efficiency of single-particle spectro-microscopy analysis using the spectra phasor approach when compared to a conventional optical grating method was demonstrated.
Optical microscopy has become an invaluable tool for investigating complex samples. Over the years, many advances to optical microscopes have been made that have allowed us to uncover new insights into the samples studied. Dynamic changes in biological and chemical systems are of utmost importance to study. To probe these samples, multidimensional approaches have been developed to acquire a fuller understanding of the system of interest. These dimensions include the spatial information, such as the three-dimensional coordinates and orientation of the optical probes, and additional chemical and physical properties through combining microscopy with various spectroscopic techniques. In this review, we survey the field of multidimensional microscopy and provide an outlook on the field and challenges that may arise. Expected final online publication date for the Annual Review of Physical Chemistry, Volume 73 is April 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.
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