Brain circuits are highly interconnected three-dimensional structures fabricated from components ranging vastly in size; from cell bodies to individual synapses. While neuronal activity can be visualized with advanced light microscopy (LM) techniques, the resolution of electron microscopy (EM) is critical for identifying synaptic connections between neurons. Here, we combine these two techniques, affording the advantage of each and allowing for measurements to be made of the same neural features across imaging platforms. We established an EM-label-free workflow utilizing inherent structural features to correlate in vivo two-photon LM and volumetric scanning EM (SEM) in the ferret visual cortex. By optimizing the volume SEM sample preparation protocol, imaging with the OnPoint detector, and utilizing the focal charge compensation device during serial block-face imaging, we achieved sufficient resolution and signal-to-noise ratio to analyze synaptic ultrastructure for hundreds of synapses within sample volumes. Our novel workflow provides a reliable method for quantitatively characterizing synaptic ultrastructure in functionally imaged neurons, providing new insights into neuronal circuit organization.
You may also like
Insulin-like hormones critical for brain plasticity
August 7, 2023Max Planck Florida Institute for Neuroscience
A Butterfly Effect
July 27, 2023Max Planck Florida Institute for Neuroscience
Deep learning models to study sentence comprehension...
June 28, 2023Max Planck Institute for Psycholinguistics
How the brain slows down when we focus our gaze
June 28, 2023Max Planck Institute for Biological Cybernetics
Fruit fly’s complex symphony of vision
June 6, 2023Max Planck Institute for Biological Intelligence
How tasty is the food? Ask your brain!
June 6, 2023Max Planck Institute for Biological Intelligence