Studying synapses in human brain with array tomography and electron microscopy

Kevin R Kay; Colin Smith; Ann K Wright; Alberto Serrano-Pozo; Amy M Pooler; Robert Koffie; Mark E Bastin; Thomas H Bak; Sharon Abrahams; Katherine J Kopeikina; Declan McGuone; Matthew P Frosch; Thomas H Gillingwater; Bradley T Hyman; Tara L Spires-Jones

doi:10.1038/nprot.2013.078

Studying synapses in human brain with array tomography and electron microscopy

Kevin R Kay, Colin Smith, Ann K Wright, Alberto Serrano-Pozo, Amy M Pooler, Robert Koffie, Mark E Bastin, Thomas H Bak, Sharon Abrahams, Katherine J Kopeikina, Declan McGuone, Matthew P Frosch, Thomas H Gillingwater, Bradley T Hyman, Tara L Spires-Jones

Research output: Contribution to journal › Article › peer-review

81 Citations (Scopus)

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.

Original language	English
Pages (from-to)	1366-1380
Number of pages	15
Journal	Nature Protocols
Volume	8
Issue number	7
DOIs	https://doi.org/10.1038/nprot.2013.078
Publication status	Published - 20 Jun 2013

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1038/nprot.2013.078

Cite this

Kay, K. R., Smith, C., Wright, A. K., Serrano-Pozo, A., Pooler, A. M., Koffie, R., Bastin, M. E., Bak, T. H., Abrahams, S., Kopeikina, K. J., McGuone, D., Frosch, M. P., Gillingwater, T. H., Hyman, B. T., & Spires-Jones, T. L. (2013). Studying synapses in human brain with array tomography and electron microscopy. Nature Protocols, 8(7), 1366-1380. https://doi.org/10.1038/nprot.2013.078

@article{f01149706b1d420fb17c34b2ae6662b4,

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.",

author = "Kay, {Kevin R} and Colin Smith and Wright, {Ann K} and Alberto Serrano-Pozo and Pooler, {Amy M} and Robert Koffie and Bastin, {Mark E} and Bak, {Thomas H} and Sharon Abrahams and Kopeikina, {Katherine J} and Declan McGuone and Frosch, {Matthew P} and Gillingwater, {Thomas H} and Hyman, {Bradley T} and Spires-Jones, {Tara L}",

year = "2013",

month = jun,

day = "20",

doi = "10.1038/nprot.2013.078",

language = "English",

volume = "8",

pages = "1366--1380",

journal = "Nature Protocols",

issn = "1754-2189",

publisher = "Springer Nature",

number = "7",

}

TY - JOUR

T1 - Studying synapses in human brain with array tomography and electron microscopy

AU - Kay, Kevin R

AU - Smith, Colin

AU - Wright, Ann K

AU - Serrano-Pozo, Alberto

AU - Pooler, Amy M

AU - Koffie, Robert

AU - Bastin, Mark E

AU - Bak, Thomas H

AU - Abrahams, Sharon

AU - Kopeikina, Katherine J

AU - McGuone, Declan

AU - Frosch, Matthew P

AU - Gillingwater, Thomas H

AU - Hyman, Bradley T

AU - Spires-Jones, Tara L

PY - 2013/6/20

Y1 - 2013/6/20

N2 - 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.

AB - 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.

U2 - 10.1038/nprot.2013.078

DO - 10.1038/nprot.2013.078

M3 - Article

C2 - 23787894

SN - 1754-2189

VL - 8

SP - 1366

EP - 1380

JO - Nature Protocols

JF - Nature Protocols

IS - 7

ER -

Studying synapses in human brain with array tomography and electron microscopy

Abstract

UN SDGs

Access to Document

Fingerprint

Cite this