Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals
Abstract 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...
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| Format: | Article |
| Language: | English |
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BMC
2019-09-01
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| Series: | Acta Neuropathologica Communications |
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| Online Access: | http://link.springer.com/article/10.1186/s40478-019-0794-3 |
| _version_ | 1828402043635630080 |
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| author | Mariella Sele Stefan Wernitznig Saška Lipovšek Snježana Radulović Johannes Haybaeck Anna Maria Birkl-Toeglhofer Christina Wodlej Florian Kleinegger Stephan Sygulla Marlene Leoni Stefan Ropele Gerd Leitinger |
| author_facet | Mariella Sele Stefan Wernitznig Saška Lipovšek Snježana Radulović Johannes Haybaeck Anna Maria Birkl-Toeglhofer Christina Wodlej Florian Kleinegger Stephan Sygulla Marlene Leoni Stefan Ropele Gerd Leitinger |
| author_sort | Mariella Sele |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-10T09:55:02Z |
| format | Article |
| id | doaj.art-45c61f37eaf14e6da6db0071f314161f |
| institution | Directory Open Access Journal |
| issn | 2051-5960 |
| language | English |
| last_indexed | 2024-12-10T09:55:02Z |
| publishDate | 2019-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Acta Neuropathologica Communications |
| spelling | doaj.art-45c61f37eaf14e6da6db0071f314161f2022-12-22T01:53:32ZengBMCActa Neuropathologica Communications2051-59602019-09-017111210.1186/s40478-019-0794-3Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervalsMariella Sele0Stefan Wernitznig1Saška Lipovšek2Snježana Radulović3Johannes Haybaeck4Anna Maria Birkl-Toeglhofer5Christina Wodlej6Florian Kleinegger7Stephan Sygulla8Marlene Leoni9Stefan Ropele10Gerd Leitinger11Research Unit Electron Microscopic Techniques, Division of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of GrazResearch Unit Electron Microscopic Techniques, Division of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of GrazResearch Unit Electron Microscopic Techniques, Division of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of GrazResearch Unit Electron Microscopic Techniques, Division of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of GrazDiagnostic and Research Institute of Pathology, Diagnostic & Research Center for Molecular BioMedicine, Medical University of GrazDiagnostic and Research Institute of Pathology, Diagnostic & Research Center for Molecular BioMedicine, Medical University of GrazDiagnostic and Research Institute of Pathology, Diagnostic & Research Center for Molecular BioMedicine, Medical University of GrazDiagnostic and Research Institute of Pathology, Diagnostic & Research Center for Molecular BioMedicine, Medical University of GrazDiagnostic and Research Institute of Pathology, Diagnostic & Research Center for Molecular BioMedicine, Medical University of GrazDiagnostic and Research Institute of Pathology, Diagnostic & Research Center for Molecular BioMedicine, Medical University of GrazDepartment of Neurology, Medical University of GrazResearch Unit Electron Microscopic Techniques, Division of Cell Biology, Histology and Embryology, Gottfried Schatz Research Center, Medical University of GrazAbstract 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.http://link.springer.com/article/10.1186/s40478-019-0794-3Electron microscopyHigh pressure freezingFreeze substitutionPost-mortem intervalHuman brain |
| spellingShingle | Mariella Sele Stefan Wernitznig Saška Lipovšek Snježana Radulović Johannes Haybaeck Anna Maria Birkl-Toeglhofer Christina Wodlej Florian Kleinegger Stephan Sygulla Marlene Leoni Stefan Ropele Gerd Leitinger Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals Acta Neuropathologica Communications Electron microscopy High pressure freezing Freeze substitution Post-mortem interval Human brain |
| title | Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals |
| title_full | Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals |
| title_fullStr | Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals |
| title_full_unstemmed | Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals |
| title_short | Optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post-mortem intervals |
| title_sort | optimization of ultrastructural preservation of human brain for transmission electron microscopy after long post mortem intervals |
| topic | Electron microscopy High pressure freezing Freeze substitution Post-mortem interval Human brain |
| url | http://link.springer.com/article/10.1186/s40478-019-0794-3 |
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