Fluorescence nanoscopy in cell biology

Abstract

Fluorescence nanoscopy uniquely combines minimally invasive optical access to the internal nanoscale structure and dynamics of cells and tissues with molecular detection specificity. While the basic physical principles of 'super-resolution' imaging were discovered in the 1990s, with initial experimental demonstrations following in 2000, the broad application of super-resolution imaging to address cell-biological questions has only more recently emerged. Nanoscopy approaches have begun to facilitate discoveries in cell biology and to add new knowledge. One current direction for method improvement is the ambition to quantitatively account for each molecule under investigation and assess true molecular colocalization patterns via multi-colour analyses. In pursuing this goal, the labelling of individual molecules to enable their visualization has emerged as a central challenge. Extending nanoscale imaging into (sliced) tissue and whole-animal contexts is a further goal. In this Review we describe the successes to date and discuss current obstacles and possibilities for further development.

The Spectrin-Actin-Based Periodic Cytoskeleton as a Conserved Nanoscale Scaffold and Ruler of the Neural Stem Cell Lineage

“…The recent discovery (Xu et al, 2013) of a highly structured and periodic membrane cytoskeleton in neurons via super-resolution microscopy (SRM) (Huang et al, 2010; Sahl et al, 2017) has kindled great interest in the ultrastructure of the membrane cytoskeleton in cells of the nervous system (Albrecht et al, 2016; Bär et al, 2016; D’Este et al, 2015, 2016, 2017; Ganguly et al, 2015; Han et al, 2017; He et al, 2016; Leite et al, 2016; Leterrier et al, 2015, 2017; Sidenstein et al, 2016; Xu et al, 2013; Zhong et al, 2014). Although initially noted in neuronal axons as adducin-capped actin rings connected by spectrin tetramers to form a periodic, one-dimensional (1D) lattice of well-defined, ~180- to 190-nm periodicity (Xu et al, 2013), related periodic or quasi-periodic cytoskeletal structures have also been observed in dendrites (D’Este et al, 2015; Han et al, 2017) and certain glial cell types (D’Este et al, 2016, 2017; He et al, 2016).…”

Nanoscale colocalization of NK cell activating and inhibitory receptors controls signal integration

“…microclusters [32,45,46]. Following the development of SR microscopy, we are now starting to have an insight of the nanometer scale organization of immune receptors at the cell surface [47,48]. However, the use of SR techniques poses some challenges e.g.…”

Histochemistry today: Detection and location of single molecules

“…Consequently, in biomedical research it is crucial to specifically label and localize molecules there, where they exert their structural roles and activities. Thanks to the progress of super-resolution microscopy, it is nowadays possible to obtain multicolor fluorescence images at the nanoscale, with a resolution comparable to electron microscopy: [101][102][103] no doubt, histochemistry will continue to provide scientists the most appropriate tools for precisely tracing molecular maps in the attempt to get a mechanistic explanation of the "behaviour" …”