Evaluating ultrastructural preservation quality in banked brain tissue

The ultrastructural analysis of postmortem brain tissue can provide important insights into cellular architecture and disease-related changes. For example, connectomics studies offer a powerful emerging approach for understanding neural circuit organization. However, electron microscopy (EM) data i...

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Main Authors: Macy Garrood, Alicia Keberle, Allison Sowa, William Janssen, Emma Thorn, Claudia De Sanctis, Kurt Farrell, John Crary, Andrew McKenzie
Format: Article
Language:English
Published: University of Münster / Open Journals System 2025-06-01
Series:Free Neuropathology
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Online Access:https://www.uni-muenster.de/Ejournals/index.php/fnp/article/view/6763
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author Macy Garrood
Alicia Keberle
Allison Sowa
William Janssen
Emma Thorn
Claudia De Sanctis
Kurt Farrell
John Crary
Andrew McKenzie
author_facet Macy Garrood
Alicia Keberle
Allison Sowa
William Janssen
Emma Thorn
Claudia De Sanctis
Kurt Farrell
John Crary
Andrew McKenzie
author_sort Macy Garrood
collection DOAJ
description The ultrastructural analysis of postmortem brain tissue can provide important insights into cellular architecture and disease-related changes. For example, connectomics studies offer a powerful emerging approach for understanding neural circuit organization. However, electron microscopy (EM) data is difficult to interpret when the preservation quality is imperfect, which is common in brain banking and may render it unsuitable for certain research applications. One common issue is that EM images of postmortem brain tissue can have an expansion of regions that appear to be made up of extracellular space and / or degraded cellular material, which we call ambiguous interstitial zones. In this study, we report a method to assess whether EM images have ambiguous interstitial zone artifacts in a cohort of 10 postmortem brains with samples from each of the cortex and thalamus. Next, in matched samples from the contralateral hemisphere of the same brains, we evaluate the structural preservation quality of light microscopy images, including immunostaining for cytoskeletal proteins. Through this analysis, we show that on light microscopy, cell membrane morphology can be largely maintained, and neurite trajectory visualized over micrometer distances, even in specimens for which there are ambiguous interstitial zone artifacts on EM. Additionally, we demonstrate that synaptic structures can be successfully traced across serial EM sections in some postmortem samples, indicating the potential for connectivity studies in banked human brain tissue when appropriate preservation and visualization protocols are employed. Taken together, our analysis may assist in maximizing the usefulness of donated brain tissue by informing tissue selection and preparation protocols for various research goals.
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spelling doaj-art-9c61219f351348b286eeb78d1dceec702025-08-20T03:32:11ZengUniversity of Münster / Open Journals SystemFree Neuropathology2699-44452025-06-01610.17879/freeneuropathology-2025-6763Evaluating ultrastructural preservation quality in banked brain tissueMacy Garrood0Alicia Keberle1Allison Sowa2William Janssen3Emma Thorn4Claudia De Sanctis5Kurt Farrell6John Crary7Andrew McKenzie8Apex Neuroscience, Salem, Oregon, USAApex Neuroscience, Salem, Oregon, USAMicroscopy and Advanced Bioimaging Core, Icahn School of Medicine at Mount Sinai, New York, New York, USAMicroscopy and Advanced Bioimaging Core, Icahn School of Medicine at Mount Sinai, New York, New York, USAFriedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USAFriedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USAFriedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USAFriedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USAApex Neuroscience, Salem, Oregon, USA The ultrastructural analysis of postmortem brain tissue can provide important insights into cellular architecture and disease-related changes. For example, connectomics studies offer a powerful emerging approach for understanding neural circuit organization. However, electron microscopy (EM) data is difficult to interpret when the preservation quality is imperfect, which is common in brain banking and may render it unsuitable for certain research applications. One common issue is that EM images of postmortem brain tissue can have an expansion of regions that appear to be made up of extracellular space and / or degraded cellular material, which we call ambiguous interstitial zones. In this study, we report a method to assess whether EM images have ambiguous interstitial zone artifacts in a cohort of 10 postmortem brains with samples from each of the cortex and thalamus. Next, in matched samples from the contralateral hemisphere of the same brains, we evaluate the structural preservation quality of light microscopy images, including immunostaining for cytoskeletal proteins. Through this analysis, we show that on light microscopy, cell membrane morphology can be largely maintained, and neurite trajectory visualized over micrometer distances, even in specimens for which there are ambiguous interstitial zone artifacts on EM. Additionally, we demonstrate that synaptic structures can be successfully traced across serial EM sections in some postmortem samples, indicating the potential for connectivity studies in banked human brain tissue when appropriate preservation and visualization protocols are employed. Taken together, our analysis may assist in maximizing the usefulness of donated brain tissue by informing tissue selection and preparation protocols for various research goals. https://www.uni-muenster.de/Ejournals/index.php/fnp/article/view/6763Brain bankingPostmortem changesPerfusion fixationNeurofilamentsUltrastructural qualityConnectomics
spellingShingle Macy Garrood
Alicia Keberle
Allison Sowa
William Janssen
Emma Thorn
Claudia De Sanctis
Kurt Farrell
John Crary
Andrew McKenzie
Evaluating ultrastructural preservation quality in banked brain tissue
Free Neuropathology
Brain banking
Postmortem changes
Perfusion fixation
Neurofilaments
Ultrastructural quality
Connectomics
title Evaluating ultrastructural preservation quality in banked brain tissue
title_full Evaluating ultrastructural preservation quality in banked brain tissue
title_fullStr Evaluating ultrastructural preservation quality in banked brain tissue
title_full_unstemmed Evaluating ultrastructural preservation quality in banked brain tissue
title_short Evaluating ultrastructural preservation quality in banked brain tissue
title_sort evaluating ultrastructural preservation quality in banked brain tissue
topic Brain banking
Postmortem changes
Perfusion fixation
Neurofilaments
Ultrastructural quality
Connectomics
url https://www.uni-muenster.de/Ejournals/index.php/fnp/article/view/6763
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