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Astrocytes pp 69-84 | Cite as

A Method to Visualize the Nanoscopic Morphology of Astrocytes In Vitro and In Situ

  • Janosch P. HellerEmail author
  • Dmitri A. RusakovEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1938)

Abstract

In recent years it has become apparent that astroglia are not only essential players in brain development, homeostasis, and metabolic support but are also important for the formation and regulation of synaptic circuits. Fine astrocytic processes that can be found in the vicinity of synapses undergo considerable structural plasticity associated with age- and use-dependent changes in neural circuitries. However, due to the extraordinary complex, essentially nanoscopic morphology of astroglia, the underlying cellular mechanisms remain poorly understood.

Here we detail a super-resolution microscopy approach, based on the single-molecule localisation microscopy (SMLM) technique direct stochastic optical reconstruction microscopy (dSTORM) to visualize astroglial morphology on the nanoscale. This approach enables visualization of key morphological changes that occur in nanoscopic astrocyte processes, whose characteristic size falls below the diffraction limit of conventional optical microscopy.

Key words

Super-resolution microscopy SMLM dSTORM Brain tissue fixation Tissue sectioning Immunohistochemistry Astrocytes 

Notes

Acknowledgments

The authors thank Dr. Piotr Michaluk for cell culture preparations. This research was supported by Wellcome Trust Principal Fellowship (101896), European Research Council Advanced Grant (323113-NETSIGNAL), FP7 ITN (606950 EXTRABRAIN), and Russian Science Foundation grant (15-14-30000).

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.UCL Queen Square Institute of NeurologyUniversity College London, Queen SquareLondonUK
  2. 2.Laboratory of Brain Microcircuits, Institute of NeuroscienceUniversity of Nizhny NovgorodNizhny NovgorodRussia

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