title: Studying synapses in human brain with array tomography and electron microscopy.
abstract: Postmortem studies of synapses in human brain are problematic because of the axial resolution limit of light microscopy and the difficulty in preserving and analyzing ultrastructure with electron microscopy (EM). Array tomography (AT) overcomes these problems by embedding autopsy tissue in resin and cutting ribbons of ultrathin serial sections. Ribbons are imaged with immunofluorescence, allowing high-throughput imaging of tens of thousands of synapses to assess synapse density and protein composition. The protocol takes ~3 d per case, excluding image analysis, which is done at the end of the study. Parallel processing for transmission electron microscopy (TEM) using a protocol modified to preserve the structure in human samples allows complementary ultrastructural studies. Incorporation of AT and TEM into brain banking is a potent way of phenotyping synapses in well-characterized clinical cohorts in order to develop clinicopathological correlations at the synapse level. This will be important for research in neurodegenerative disease, developmental disease and psychiatric illness.
authors.affiliation: Massachusetts General Hospital and Harvard Medical School, MassGeneral Institute for Neurodegenerative Disease, Charlestown, MA, USA.
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Kay
authors.forename: Colin
authors.initials: C
authors.lastname: Smith
authors.forename: Ann K
authors.initials: AK
authors.lastname: Wright
authors.forename: Alberto
authors.initials: A
authors.lastname: Serrano-Pozo
authors.forename: Amy M
authors.initials: AM
authors.lastname: Pooler
authors.forename: Robert
authors.initials: R
authors.lastname: Koffie
authors.forename: Mark E
authors.initials: ME
authors.lastname: Bastin
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Bak
authors.forename: Sharon
authors.initials: S
authors.lastname: Abrahams
authors.forename: Katherine J
authors.initials: KJ
authors.lastname: Kopeikina
authors.forename: Declan
authors.initials: D
authors.lastname: McGuone
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Frosch
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Gillingwater
authors.forename: Bradley T
authors.initials: BT
authors.lastname: Hyman
authors.forename: Tara L
authors.initials: TL
authors.lastname: Spires-Jones
mesh_terms: D019412:Anatomy, Cross-Sectional / Q000379:methods; D001344:Autopsy; D001921:Brain / Q000166:cytology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D013569:Synapses / Q000648:ultrastructure*; D014054:Tomography / Q000379:methods*