Extended Data Fig. 1: Correlating in vivo synaptic imaging and serial block-face scanning electron microscopy.
From: Cortical response selectivity derives from strength in numbers of synapses
a, In vivo two-photon synaptic imaging of L2/3 cortical neurons in ferret visual cortex expressing GCaMP6s is performed under visual stimulation. Imaged cells are identified after perfusion and re-imaged with a confocal microscope. Tissue is trimmed and processed for serial-block-face scanning electron microscopy (SBF-SEM). Imaged cells are identified within the block of tissue via biological fiducials and high-resolution SEM is performed. Finally, imaged cells, dendritic spines, and corresponding synaptic features are volumetrically reconstructed for quantification. Dendrite is shown in brown, spine neck and head are shown in blue, post-synaptic density is shown in red, and presynaptic bouton is shown in green.
