Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals.
Electron microscopy
Freeze substitution
High pressure freezing
Human brain
Post-mortem interval
Journal
Acta neuropathologica communications
ISSN: 2051-5960
Titre abrégé: Acta Neuropathol Commun
Pays: England
ID NLM: 101610673
Informations de publication
Date de publication:
03 09 2019
03 09 2019
Historique:
received:
01
07
2019
accepted:
24
08
2019
entrez:
5
9
2019
pubmed:
5
9
2019
medline:
24
6
2020
Statut:
epublish
Résumé
Electron microscopy (EM) provides the necessary resolution to visualize the finer structures of nervous tissue morphology, which is important to understand healthy and pathological conditions in the brain. However, for the interpretation of the micrographs the tissue preservation is crucial. The quality of the tissue structure is mostly influenced by the post mortem interval (PMI), the time of death until the preservation of the tissue. Therefore, the aim of this study was to optimize the preparation-procedure for the human frontal lobe to preserve the ultrastructure as well as possible despite the long PMIs. Combining chemical pre- and post-fixation with cryo-fixation and cryo-substitution ("hybrid freezing"), it was possible to improve the preservation of the neuronal profiles of human brain samples compared to the "standard" epoxy resin embedding method. In conclusion short PMIs are generally desirable but up to a PMI of 16 h the ultrastructure can be preserved on an acceptable level with a high contrast using the "hybrid freezing" protocol described here.
Identifiants
pubmed: 31481118
doi: 10.1186/s40478-019-0794-3
pii: 10.1186/s40478-019-0794-3
pmc: PMC6724377
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
144Subventions
Organisme : Austrian Science Fund FWF
ID : P 29370
Pays : Austria
Organisme : Austrian Science Fund
ID : P-29370B27
Pays : International
Références
Brain Res. 2016 Jun 15;1641(Pt A):92-100
pubmed: 26920467
Cell Biophys. 1991 Feb;18(1):31-55
pubmed: 1725502
Elife. 2018 May 11;7:
pubmed: 29749931
J Biophys Biochem Cytol. 1956 Sep 25;2(5):531-42
pubmed: 13376632
Brain Res. 1997 Feb 21;749(1):82-7
pubmed: 9070630
J Microsc. 2003 Oct;212(Pt 1):3-12
pubmed: 14516356
Methods Cell Biol. 2007;79:7-21
pubmed: 17327150
Ann Anat. 2016 Jul;206:27-33
pubmed: 27136748
J Struct Biol. 2005 May;150(2):144-53
pubmed: 15866737
Acta Neuropathol Commun. 2018 Mar 2;6(1):22
pubmed: 29499767
Traffic. 2018 Aug;19(8):639-649
pubmed: 29673018
Sci Rep. 2018 Mar 5;8(1):3979
pubmed: 29507421
Protist. 2016 Aug;167(4):369-376
pubmed: 27434544
J Histochem Cytochem. 2018 Dec;66(12):903-921
pubmed: 29969056
Neurobiol Aging. 1995 May-Jun;16(3):271-8; discussion 278-84
pubmed: 7566337
J Histochem Cytochem. 2019 Mar;67(3):185-202
pubmed: 30562121
Nature. 2019 Feb;566(7745):538-542
pubmed: 30675058
Acta Neuropathol Commun. 2018 Oct 1;6(1):102
pubmed: 30270816
Protoplasma. 2014 Mar;251(2):429-48
pubmed: 24258967
Methods Cell Biol. 2010;96:475-512
pubmed: 20869535
Ann Anat. 2009 Jun;191(3):231-47
pubmed: 19264467
Brain Struct Funct. 2016 Jun;221(5):2477-86
pubmed: 25905864
J Struct Biol. 2008 Mar;161(3):359-71
pubmed: 17962040
Acta Neuropathol Commun. 2015 Sep 04;3:53
pubmed: 26335101
Methods Cell Biol. 2010;96:671-93
pubmed: 20869543
Histochem Cell Biol. 2008 Nov;130(5):877-89
pubmed: 18795316
Nat Protoc. 2013;8(7):1366-80
pubmed: 23787894
Biotechniques. 2006 Aug;41(2):137, 139, 141 passim
pubmed: 16925014
Elife. 2015 Aug 11;4:
pubmed: 26259873
Mech Ageing Dev. 1980 Sep-Oct;14(1-2):211-22
pubmed: 6163044
Acta Pharmacol Sin. 2011 Sep;32(9):1089-99
pubmed: 21804578
J Microsc. 2008 May;230(Pt 2):268-77
pubmed: 18445157
J Microsc. 1984 May;134(Pt 2):213-6
pubmed: 6429337
Methods Cell Biol. 2007;79:23-56
pubmed: 17327151
Acta Neuropathol Commun. 2014 Apr 11;2:42
pubmed: 24721148
Cell Signal. 2011 Oct;23(10):1534-45
pubmed: 21683788