title: Structured Illumination Microscopy for the Investigation of Synaptic Structure and Function.
abstract: 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.
authors.affiliation: Department of Neurology, F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.|Program in Neuroscience, Harvard Medical School, Boston, MA, USA.
authors.forename: Soyon
authors.initials: S
authors.lastname: Hong
authors.affiliation: Department of Neurology, F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.|Program in Neuroscience, Harvard Medical School, Boston, MA, USA.
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Wilton
authors.affiliation: Department of Neurology, F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.|Program in Neuroscience, Harvard Medical School, Boston, MA, USA.
authors.forename: Beth
authors.initials: B
authors.lastname: Stevens
authors.affiliation: Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA, USA. drichardson@fas.harvard.edu.|Harvard Center for Biological Imaging, Harvard University, 16 Divinity Avenue, Cambridge, MA, 02138, USA. drichardson@fas.harvard.edu.
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Richardson
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000000981:diagnostic imaging / Q000378:metabolism; D021621:Imaging, Three-Dimensional / Q000379:methods; D051379:Mice; D008853:Microscopy / Q000379:methods*; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D013569:Synapses / Q000502:physiology*
keywords: Channel alignment; Colocalization; Immunofluorescence; Neuron; Refractive index matching; Structured Illumination Microscopy; Synapse