Insights into Structure, Dynamics, and Function of Endocytic Vesicles on the STELLARIS Confocal Platform

Imaging live cells is key to reveal their inner workings, dynamics, and function. Current technological advances in confocal imaging can benefit from novel fluorescence labeling strategies and biosensors. It is possible to probe biologically relevant processes faster and easier than before. Still, understanding the underlying mechanisms of any given process requires the observation of a specimen from different angles. Intensity-based imaging provides a reliable view of the spatial organization and distribution of intracellular species and organelles, whereas lifetime-based approaches can elicit information from the micro-environment and other functional related signals. Moreover, fluorescence nanoscopies have evolved into the go-to technique to examine structural details beyond the physical diffraction limits, with molecular specificity, and within the crowded cellular milieu.

 

In this talk we will show how STELLARIS provides an integral access to these different layers of information from dynamic, structural, and mechanistic insights into vesicle trafficking. The endocytic vesicles shuttle materials between different cellular compartments, with the extracellular space, and the endocytic pathway is vital to almost all aspects of cell life and disease. This pathway is of particular interest in host-pathogen studies as many viruses and bacteria can exploit it as an cell entry point.

 

We will discuss the main aspects of our confocal platform that enable excellent image quality, spatiotemporal resolution even at the nanoscopy level, and functional information. We will explain the operating principles behind Power Counting to significantly improve image contrast while delivering quantitative results, and how the combination with the AI-based Aivia tools allows characterizing the dynamics of the vesicle trafficking. To access functional information, we will focus on TauSense technology, a new, straightforward way to access lifetime-based information in confocal imaging.

Learning Objectives