Abstract
Connectomics allows mapping of cells and their circuits at the nanometer scale in
volumes of approximately 1 mm3. Given that the human cerebral cortex can be 3 mm in thickness, larger volumes are
required. Larger-volume circuit reconstructions of human brain are limited by 1) the
availability of fresh biopsies; 2) the need for excellent preservation of ultrastructure,
including extracellular space; and 3) the requirement of uniform staining throughout
the sample, among other technical challenges. Cerebral cortical samples from neurosurgical
patients are available owing to lead placement for deep brain stimulation. Described
here is an immersion fixation, heavy metal staining, and tissue processing method
that consistently provides excellent ultrastructure throughout human and rodent surgical
brain samples of volumes 2 × 2 × 2 mm3 and up to 37 mm3 with one dimension ≤2 mm. This method should allow synapse-level circuit analysis
in samples from patients with psychiatric and neurologic disorders.
Keywords
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Article info
Publication history
Published online: February 03, 2023
Accepted:
January 27,
2023
Received in revised form:
January 25,
2023
Received:
August 6,
2022
Handling Editor: Dr. J.R. KrystalPublication stage
In Press Journal Pre-ProofIdentification
Copyright
© 2023 Society of Biological Psychiatry.
