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Methods Mol Biol DOI:10.1007/978-1-4939-6688-2_12

Structured Illumination Microscopy for the Investigation of Synaptic Structure and Function.

Publication TypeJournal Article
Year of Publication2017
AuthorsHong, S, Wilton, DK, Stevens, B, Richardson, DS
JournalMethods Mol Biol
Date Published2017
KeywordsAnimals, Brain, Imaging, Three-Dimensional, Mice, Microscopy, Neurons, Synapses

The neuronal synapse is a primary building block of the nervous system to which alterations in structure or function can result in numerous pathologies. Studying its formation and elimination is the key to understanding how brains are wired during development, maintained throughout adulthood plasticity, and disrupted during disease. However, due to its diffraction-limited size, investigations of the synaptic junction at the structural level have primarily relied on labor-intensive electron microscopy or ultra-thin section array tomography. Recent advances in the field of super-resolution light microscopy now allow researchers to image synapses and associated molecules with high-spatial resolution, while taking advantage of the key characteristics of light microscopy, such as easy sample preparation and the ability to detect multiple targets with molecular specificity. One such super-resolution technique, Structured Illumination Microscopy (SIM), has emerged as an attractive method to examine synapse structure and function. SIM requires little change in standard light microscopy sample preparation steps, but results in a twofold improvement in both lateral and axial resolutions compared to widefield microscopy. The following protocol outlines a method for imaging synaptic structures at resolutions capable of resolving the intricacies of these neuronal connections.


Alternate JournalMethods Mol. Biol.
PubMed ID27943190
PubMed Central IDPMC5479421
Grant ListR01 NS071008 / NS / NINDS NIH HHS / United States
S10 RR029237 / RR / NCRR NIH HHS / United States
T32 AG000222 / AG / NIA NIH HHS / United States
U54 HD090255 / HD / NICHD NIH HHS / United States