Superresolution imaging of chemical synapses in the brain

Neuron. 2010 Dec 9;68(5):843-56. doi: 10.1016/j.neuron.2010.11.021.


Determination of the molecular architecture of synapses requires nanoscopic image resolution and specific molecular recognition, a task that has so far defied many conventional imaging approaches. Here, we present a superresolution fluorescence imaging method to visualize the molecular architecture of synapses in the brain. Using multicolor, three-dimensional stochastic optical reconstruction microscopy, the distributions of synaptic proteins can be measured with nanometer precision. Furthermore, the wide-field, volumetric imaging method enables high-throughput, quantitative analysis of a large number of synapses from different brain regions. To demonstrate the capabilities of this approach, we have determined the organization of ten protein components of the presynaptic active zone and the postsynaptic density. Variations in synapse morphology, neurotransmitter receptor composition, and receptor distribution were observed both among synapses and across different brain regions. Combination with optogenetics further allowed molecular events associated with synaptic plasticity to be resolved at the single-synapse level.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism
  • Brain / ultrastructure*
  • Brain Chemistry / physiology
  • Image Processing, Computer-Assisted / instrumentation
  • Image Processing, Computer-Assisted / methods
  • Imaging, Three-Dimensional / instrumentation
  • Imaging, Three-Dimensional / methods*
  • Immunohistochemistry / methods
  • Mice
  • Mice, Inbred C57BL
  • Microchemistry / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Molecular Imaging / instrumentation
  • Molecular Imaging / methods
  • Nanotechnology
  • Nerve Tissue Proteins / metabolism
  • Nerve Tissue Proteins / ultrastructure*
  • Neuronal Plasticity / physiology
  • Photomicrography / instrumentation
  • Photomicrography / methods
  • Presynaptic Terminals / metabolism
  • Presynaptic Terminals / ultrastructure*
  • Stochastic Processes
  • Synapses / metabolism
  • Synapses / ultrastructure


  • Nerve Tissue Proteins