Parte de libro
Single-molecule localization super-resolution microscopy of synaptic proteins
Registro en:
978-1-4939-6836-7
978-1-4939-6836-7 (online)
1949-2448
10.1007/8623_2016_10
Autor
Barrantes, Francisco José
Institución
Resumen
Abstract: Recent years have witnessed huge progress in the field of light microscopy with the development and
implementation of new approaches leading to dramatic improvements in the spatial and temporal resolution
of this form of imaging, most particularly in its biological applications. The limitations in spatial
resolution imposed by the diffraction of light have been circumvented by resorting to different strategies,
which are briefly outlined in the Introduction. These protocols are intended to provide practical guidelines
for the imaging of synaptic proteins using one such strategy, namely, single-molecule stochastic localization
super-resolution microscopy.
The protocols use neuronal cells from the hippocampus of rodent embryos as the experimental paradigm
and outline the steps for obtaining dissociated neurons and establishing primary cultures for in vitro studies.
The techniques can be adapted to the culture of neurons from other brain regions. Procedures for handling
fixed and live specimens are described, as well as the use of extrinsic fluorescent probes and fluorescent
proteins, mounting media, examples of hardware configurations, software for image analysis, and some
hints for the implementation of minimalist approaches to single-molecule localization nanoscopy.