Array Tomography for Cortical Tissue Analysis

Array tomography analyses were performed as previously described (Hudry et al, 2013 (link); Kay et al, 2013 (link)). Five to six pieces of cortical tissue (1 mm³) were dissected and fixed for 3 h in 4% paraformaldehyde and 2.5% sucrose in 0.01 M PBS. After dehydration in ethanol, the samples were incubated in LR White resin (Electron Microscopy Sciences) overnight at 4°C before polymerization at 53°C. Ribbons of 20–40 ultrathin (70 nm thick) serial sections were cut with a Histo Jumbo diamond knife (Diatome) on an ultracut microtome (Leica) and mounted on glass coverslips.

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Hudry E., Klickstein J., Cannavo C., Jackson R., Muzikansky A., Gandhi S., Urick D., Sargent T., Wrobleski L., Roe A.D., Hou S.S., Kuchibhotla K.V., Betensky R.A., Spires-Jones T, & Hyman B.T. (2019). Opposing Roles of apolipoprotein E in aging and neurodegeneration. Life Science Alliance, 2(1), e201900325.

Publication 2019

LR White resin Histo Jumbo diamond knife ultracut microtome

Cortical Dehydration ethanol Diamond Electron microscopy Lr white resin Microtome Paraformaldehyde Polymerization Sucrose Tissue Tomography

Corresponding Organization : Harvard University

Other organizations : UK Dementia Research Institute, University of Edinburgh, Johns Hopkins University

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Variable analysis

independent variables

Dissection of 5-6 pieces of cortical tissue (1 mm^3)
Fixation of the samples for 3 h in 4% paraformaldehyde and 2.5% sucrose in 0.01 M PBS
Dehydration of the samples in ethanol
Incubation of the samples in LR White resin (Electron Microscopy Sciences) overnight at 4°C
Polymerization of the samples at 53°C
Cutting of ribbons of 20-40 ultrathin (70 nm thick) serial sections with a Histo Jumbo diamond knife (Diatome) on an ultracut microtome (Leica)

dependent variables

Not explicitly mentioned

control variables

Consistent use of 0.01 M PBS for sample preparation
Consistent use of LR White resin for sample embedding
Consistent use of Histo Jumbo diamond knife and ultracut microtome for sectioning

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