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Super-Resolution Imaging of Subcortical White Matter using Stochastic Optical Reconstruction Microscopy (STORM) and Super-Resolution Optical Fluctuation Imaging (SOFI).

Hainsworth, AH; Lee, S; Foot, P; Patel, A; Poon, WW; Knight, AE (2018) Super-Resolution Imaging of Subcortical White Matter using Stochastic Optical Reconstruction Microscopy (STORM) and Super-Resolution Optical Fluctuation Imaging (SOFI). Neuropathol Appl Neurobiol, 44 (4). pp. 417-426. ISSN 1365-2990 https://doi.org/10.1111/nan.12426
SGUL Authors: Hainsworth, Atticus Henry

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Abstract

AIMS: The spatial resolution of light microscopy is limited by the wavelength of visible light (the "diffraction limit", approximately 250 nm). Resolution of sub-cellular structures, smaller than this limit, is possible with super resolution methods such as stochastic optical reconstruction microscopy (STORM) and super-resolution optical fluctuation imaging (SOFI). We aimed to resolve subcellular structures (axons, myelin sheaths and astrocytic processes) within intact white matter using STORM and SOFI. METHODS: Standard cryostat-cut sections of subcortical white matter from donated human brain tissue and from adult rat and mouse brain were labelled using standard immunohistochemical markers (neurofilament-H, myelin associated glycoprotein, GFAP). Image sequences were processed for STORM (effective pixel size 8-32 nm) and for SOFI (effective pixel size 80 nm). RESULTS: In human, rat and mouse subcortical white matter high quality images for axonal neurofilaments, myelin sheaths and filamentous astrocytic processes were obtained. In quantitative measurements, STORM consistently underestimated width of axons and astrocyte processes (compared with electron microscopy measurements). SOFI provided more accurate width measurements, though with somewhat lower spatial resolution than STORM. CONCLUSIONS: Super resolution imaging of intact cryo-cut human brain tissue is feasible. For quantitation, STORM can under-estimate diameters of thin fluorescent objects. SOFI is more robust. The greatest limitation for super-resolution imaging in brain sections is imposed by sample preparation. We anticipate that improved strategies to reduce autofluorescence and to enhance fluorophore performance will enable rapid expansion of this approach. [232 words] This article is protected by copyright. All rights reserved.

Item Type: Article
Additional Information: This is the peer reviewed version of the following article: A. H. Hainsworth, S. Lee, P. Foot, A. Patel, W. W. Poon and A. E. Knight (2018) Neuropathology and Applied Neurobiology 44, 417–426 Super‐resolution imaging of subcortical white matter using stochastic optical reconstruction microscopy (STORM) and super‐resolution optical fluctuation imaging (SOFI), which has been published in final form at http://dx.doi.org/10.1111/nan.12426. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Keywords: astrocytes, myelin, neurofilaments, super resolution microscopy, white matter, Neurology & Neurosurgery, 1103 Clinical Sciences, 1109 Neurosciences, 1702 Cognitive Science
SGUL Research Institute / Research Centre: Academic Structure > Molecular and Clinical Sciences Research Institute (MCS)
Journal or Publication Title: Neuropathol Appl Neurobiol
ISSN: 1365-2990
Language: eng
Dates:
DateEvent
15 May 2018Published
11 July 2017Published Online
6 July 2017Accepted
Publisher License: Publisher's own licence
Projects:
Project IDFunderFunder ID
ARUK PPG2014A-8Alzheimer’s Research UKhttp://dx.doi.org/10.13039/501100002283
PG146/151Alzheimer's SocietyUNSPECIFIED
20140901Alzheimer's Drug Discovery FoundationUNSPECIFIED
NMRC/CG/NUHS/2010Medical Research Councilhttp://dx.doi.org/10.13039/501100000265
PubMed ID: 28696566
Go to PubMed abstract
URI: https://openaccess.sgul.ac.uk/id/eprint/108999
Publisher's version: https://doi.org/10.1111/nan.12426

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