Accessible, multiscale imaging of synapses in situ using SEQUIN

Dr. Terrance Kummer

Assistant Professor of Neurology Washington University School of Medicine in St. Louis; St. Louis VA Medical Center

Dr. Kummer is an Assistant Professor of Neurology at Washington University School of Medicine and at the St. Louis VA Medical Center, a clinical neurointensivist, and the Director of the Neurotrauma ICU at Barnes Jewish Hospital. His research focuses on identifying and measuring key loci of cellular damage in traumatic brain injury and Alzheimer's disease, and seeking factors that explain the connections between these conditions. He is particularly interested in developing tools that help to unravel the complex neural networks that give rise to behavior and lead to neurological disability when injured, and on translational technologies that connect lab-based science with patients in the physiology laboratory of the intensive care unit. Dr. Kummer received his MD and PhD from Washington University in St. Louis and completed clinical training in Neurology and Neurocritical Care at Harvard Medical School.

Abstract

The complex microconnectivity of the mammalian brain underlies its computational abilities, and its vulnerability to injury and disease. It has been challenging to illuminate the features of this synaptic network due in part to the small size and exceptionally dense packing of its elements. I will describe a rapid and accessible super-resolution imaging and image analysis workflow--SEQUIN--that identifies, quantifies, and characterizes central synapses in animal models and in humans, and enables automated volumetric imaging of mesoscale synaptic networks. SEQUIN takes advantage of the ZEISS Airyscan system to resolve individual synaptic puncta and matches pre- with post-synaptic structures to identify and quantify synaptic loci. I will demonstrate how SEQUIN permitted us to identify delayed cortical synapse loss resulting from traumatic brain injury and neurodegeneration. SEQUIN mesoscale mapping of excitatory synapses across the hippocampus furthermore identified region-specific synaptic structural variability and vulnerability to injury.
 
What you will learn:
  • How to use an easy workflow to identify, quantify, and characterize synapses.
  • How to automate volumetric imaging of mesoscale synaptic networks.
  • How you can apply these techniques to your studies of brain health and disease.

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