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Understanding how the nuclear pore complex (NPC) is assembled is of fundamental importance to grasp the mechanisms behind its essential function and understand its role during the evolution of eukaryotes 1 – 4 . There are at least two NPC assembly pathways—one during the exit from mitosis and one during nuclear growth in interphase—but we currently lack a quantitative map of these events. Here we use fluorescence correlation spectroscopy calibrated live imaging of endogenously fluorescently tagged nucleoporins to map the changes in the composition and stoichiometry of seven major modules of the human NPC during its assembly in single dividing cells. This systematic quantitative map reveals that the two assembly pathways have distinct molecular mechanisms, in which the order of addition of two large structural components, the central ring complex and nuclear filaments are inverted. The dynamic stoichiometry data was integrated to create a spatiotemporal model of the NPC assembly pathway and predict the structures of postmitotic NPC assembly intermediates. Single-molecule calibrated live microscopy and computational modelling have revealed that human nuclear pore complex assembly takes different pathways during the exit from mitosis and during nuclear growth in interphase.

Today, air pollution is a global environmental problem. A huge amount of explosive and combustible gas emissions that negatively affect nature and human health. Gas sensors are one of the ways to prevent this impact. Several types of sensors have been developed, but the main problem with them is the high operating temperature. This leads to a decrease in reproducibility and stability over time. The aim of the work is to synthesize zinc stannate nanoparticles, study their phase composition, and modify the structure with silver nanoparticles to improve gas-sensing characteristics. This paper presents the synthesis of zinc stannate nanoparticles by the coprecipitation method and annealing at varying temperatures. A method of decorating zinc stannate with silver nanoparticles was proposed. Using XRD, it was found that a change in the annealing temperature leads to a change in the initial phase composition of the sample. Decoration with silver nanoparticles allows for increasing the sensor response of zinc stannate layers to isopropyl alcohol by 20 times. The corresponding increase in response to ethanol is 16 times.

Bioimaging has now entered the era of big data with faster-than-ever development of complex microscopy technologies leading to increasingly complex datasets. This enormous increase in data size and informational complexity within those datasets has brought with it several difficulties in terms of common and harmonized data handling, analysis, and management practices, which are currently hampering the full potential of image data being realized. Here, we outline a wide range of efforts and solutions currently being developed by the microscopy community to address these challenges on the path towards FAIR bioimaging data. We also highlight how different actors in the microscopy ecosystem are working together, creating synergies that develop new approaches, and how research infrastructures, such as Euro-BioImaging, are fostering these interactions to shape the field.

Understanding how the nuclear pore complex (NPC) assembles is of fundamental importance to grasp the mechanisms behind its essential function and understand its role during evolution of eukaryotes. While at least two NPC assembly pathways exist, one during exit from mitosis and one during nuclear growth in interphase, we currently lack a quantitative map of the molecular assembly events. Here, we use fluorescence correlation spectroscopy (FCS) calibrated live imaging of endogenously fluorescently-tagged nucleoporins to map the changes in composition and stoichiometry of seven major modules of the human NPC during its assembly in single dividing cells. This systematic quantitative map reveals that the two assembly pathways employ strikingly different molecular mechanisms, inverting the order of addition of two large structural components, the central ring complex and nuclear filaments. The dynamic stoichiometry data underpinned integrative spatiotemporal modeling of the NPC assembly pathway, predicting the structures of postmitotic NPC assembly intermediates. Competing Interest Statement The authors have declared no competing interest. Footnotes * Edits to main text; Supplemental files updated.

Individual molecular catalysts in a pseudohomogeneous microenvironment have been directly explored by single‐molecule observations. In this Chapter, we focus in this chapter on our proof‐of‐concept studies using immobilized metal‐based catalysts and single‐molecule fluorescence spectroscopy. Application of photostable fluorophores that signal chemical processes at the metal complex or substrate site is mandatory. Major outcomes of our research are as follows: 1. observation of the reversible formation of a single Cu(II) complex that opened a chemical approach to superresolution microscopy; 2. the irreversible epoxidation of a double–bond, exemplified by the landmark change of the emission color; 3. evidence for catalytic redox cycles of individual Cu(I/II) complexes.

This chapter contains sections titled: Introduction Basic Principles Methods Single Quantum Systems: Anti‐Bunching, Blinking, Anisotropy Conclusions and Perspectives Related Fields of Research References