New observations in neuroscience using superresolution microscopy

Abstract

Superresolution microscopy (SM) techniques are among the revolutionary methods for molecular and cellular observations in the 21 st century. SM techniques overcome optical limitations, and several new observations using SM lead us to expect these techniques to have a large impact on neuroscience in the near future. Several types of SM have been developed, including structured illumination microscopy (SIM), stimulated emission depletion microscopy (STED), and photoactivated localization microscopy (PALM)/stochastic optical reconstruction microscopy (STORM), each with special features. In this Minisymposium, experts in these different types of SM discuss the new structural and functional information about specific important molecules in neuroscience that has been gained with SM. Using these techniques, we have revealed novel mechanisms of endocytosis in nerve growth, fusion pore dynamics, and described quantitative new properties of excitatory and inhibitory synapses. Additional powerful techniques, including single molecule-guided Bayesian localization SM (SIMBA) and expansion microscopy (ExM), alone or combined with super-resolution observation, are also introduced in this session.

Fascin in lamellipodia contributes to cell elasticity by controlling the orientation of filamentous actin

“…Two important techniques were used: (a) super-resolution (Gustafsson, 2000;Hell & Wichmann, 1994) and (b) simultaneous recording by two detectors. A super-resolution microscope was essential for observing the actin meshwork because of the resolution limit (Biswas & Kalil, 2018;Igarashi et al, 2018;Nozumi et al, 2017). Thus, we used SIM and installed two cameras to record fascin and the actin meshwork simultaneously in living and moving cells.…”

“…Because of the resolution limit of conventional optical microscopy, it is difficult to observe actin meshwork in the lamellipodia region. Super-resolution microscopy is required to observe fine structures in the lamellipodia of the growth cones (Biswas & Kalil, 2018;Igarashi et al, 2018;Nozumi, Nakatsu, Katoh, & Igarashi, 2017).…”

MYCN overexpression is linked to significant differences in nuclear DNA organization in neuroblastoma

“…Super-resolution (SR) imaging has evolved from a tool built and used by physicists, to a commercially available imaging system that is now more broadly applied to multiple fields, especially in the past decade when it became obvious that sub-Abbe resolution of cellular structures became feasible and enhanced our knowledge of structures in cells dramatically [23][24][25]. For example, recent work used SR imaging to visualize CD13 for drug targeting in acute myeloid leukemia [26], hepatitis C infection [27], and organelle-cytoskeleton interactions [28]; neuroscience [29], plant cell biology [30], bacterial enzymatics [31], and cell biology [32,33] have all taken off since the development of SR imaging. Chromatin interactions and chromatin domains have been mapped using SR [34][35][36] and linked to epigenetic states [37].…”

A Single-Neuron: Current Trends and Future Prospects

“…Multiple variants of super-resolution microscopy, including structured illumination microscopy (SIM), stimulated emission depletion microscopy (STED), and photoactivated localization microscopy (PALM)/stochastic optical reconstruction microscopy (STORM), each with special features, have overcome the drawbacks of conventional microscopy and have helped remarkably in neuroscience to decipher mechanisms of endocytosis in nerve growth and fusion pore dynamics, and also describe quantitative new properties of excitatory and inhibitory synapses [72,73]. Though most recently, a super-resolution microscopy approach was developed to unravel the nanostructure of tripartite synapses with direct STORM (dSTORM) using conventional fluorophore-labeled antibodies.…”