STED Nanoscopy of Actin Dynamics in Synapses Deep Inside Living Brain Slices
Biophysical Journal. 2011-09-01; 101(5): 1277-1284
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1. Biophys J. 2011 Sep 7;101(5):1277-84. doi: 10.1016/j.bpj.2011.07.027.
STED nanoscopy of actin dynamics in synapses deep inside living brain slices.
Urban NT(1), Willig KI, Hell SW, Nägerl UV.
(1)Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
It is difficult to investigate the mechanisms that mediate long-term changes in
synapse function because synapses are small and deeply embedded inside brain
tissue. Although recent fluorescence nanoscopy techniques afford improved
resolution, they have so far been restricted to dissociated cells or tissue
surfaces. However, to study synapses under realistic conditions, one must image
several cell layers deep inside more-intact, three-dimensional preparations that
exhibit strong light scattering, such as brain slices or brains in vivo. Using
aberration-reducing optics, we demonstrate that it is possible to achieve
stimulated emission depletion superresolution imaging deep inside scattering
biological tissue. To illustrate the power of this novel (to our knowledge)
approach, we resolved distinct distributions of actin inside dendrites and spines
with a resolution of 60-80 nm in living organotypic brain slices at depths up to
120 μm. In addition, time-lapse stimulated emission depletion imaging revealed
changes in actin-based structures inside spines and spine necks, and showed that
these dynamics can be modulated by neuronal activity. Our approach greatly
facilitates investigations of actin dynamics at the nanoscale within functionally
intact brain tissue.
Copyright © 2011 Biophysical Society. Published by Elsevier Inc. All rights
PMID: 21889466 [Indexed for MEDLINE]