Current Microscopy Strategies to Image Fungal Vesicles: From the Intracellular Trafficking and Secretion to the Inner Structure of Isolated Vesicles.

Extracellular vesicles (EVs) are nano-sized structures that play important roles in a variety of biological processes among members of the Eukaryota domain. They have been studied since the 1940s and a broader use of different microscopy techniques to image either isolated vesicles or vesicles within the intracellular milieu (trafficking) has been limited by their nanometric size, usually below the resolution limit of most standard light microscopes. The development of genetically encoded fluorescent proteins and fluorescent probes able to switch between "on" and "off" states, as well the improvement in computer-assisted microscopy, photon detector devices, illumination designs, and imaging strategies in the late Twentieth century, boosted the use of light microscopes to provide structural and functional information at the sub-diffraction resolution, taking advantage of a nondestructive analytical probe such light, and opening new possibilities in the study of life at the nanoscale. As well, traditional and novel electron microscopy techniques have been widely used in the characterization of subcellular compartments, either isolated or in situ, providing a comprehensive understanding of their functional role in many cellular processes. Here, we present basic aspects of some of these techniques that have already been applied and their potential application to the study of fungal vesicles.

Spatiotemporal distribution of the magnetotactic multicellular prokaryote Candidatus Magnetoglobus multicellularis in a Brazilian hypersaline lagoon and in microcosms.

Candidatus Magnetoglobus multicellularis is an unusual morphotype of magnetotactic prokaryotes. These microorganisms are composed of a spherical assemblage of gram-negative prokaryotic cells capable of swimming as a unit aligned along a magnetic field. While they occur in many aquatic habitats around the world, high numbers of Ca. M. multicellularis have been detected in Araruama Lagoon, a large hypersaline lagoon near the city of Rio de Janeiro, in Brazil. Here, we report on the spatiotemporal distribution of one such population in sediments of Araruama Lagoon, including its annual distribution and its abundance compared with the total bacterial community. In microcosm experiments, Ca. M. multicellularis was unable to survive for more than 45 days: the population density gradually decreased coinciding with a shift to the upper layers of the sediment. Nonetheless, Ca. M. multicellularis was detected throughout the year in all sites studied. Changes in the population density seemed to be related to the input of organic matter as well as to salinity. The population density of Ca. M. multicellularis did not correlate with the total bacterial counts; instead, changes in the microbial community structure altered their counts in the environment.