Preservation of cellular structure via immersion fixation in brain banking
Immersing the brain in a solution containing formaldehyde is a commonly used method for preserving the structure of human brain tissue in brain banking. However, there are questions about the quality of preservation using this method, as formaldehyde takes a relatively long period of time to penetr...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
University of Münster / Open Journals System
2025-02-01
|
| Series: | Free Neuropathology |
| Subjects: | |
| Online Access: | https://unimuenster.de/Ejournals/index.php/fnp/article/view/6104 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832539999554240512 |
|---|---|
| author | Macy Garrood Emma L. Thorn Adam Goldstein Allison Sowa William Janssen Alyssa Wilson Claudia S. López Raakhee Shankar Erin S. Stempinski Kurt Farrell John F. Crary Andrew T. McKenzie |
| author_facet | Macy Garrood Emma L. Thorn Adam Goldstein Allison Sowa William Janssen Alyssa Wilson Claudia S. López Raakhee Shankar Erin S. Stempinski Kurt Farrell John F. Crary Andrew T. McKenzie |
| author_sort | Macy Garrood |
| collection | DOAJ |
| description |
Immersing the brain in a solution containing formaldehyde is a commonly used method for preserving the structure of human brain tissue in brain banking. However, there are questions about the quality of preservation using this method, as formaldehyde takes a relatively long period of time to penetrate a large organ such as the human brain. As a result, there is a critical need to determine whether immersion fixation is an adequate initial preservation method. To address this, we present exploratory histologic findings from our brain bank following the immersion fixation of hemi-sectioned brain specimens under refrigeration. Using light microscopy, we found that there was no significant change in the size of pericellular or perivascular rarefaction areas based on the postmortem interval (PMI) or on the progression from the outer (frontal cortex) to the inner (striatum) brain regions. Additionally, we did not identify any significant number of ghost cells – a state of late-stage cellular necrosis – in the light micrographs analyzed. Using transmission electron microscopy of tissue from the frontal cortex, we found that synapses could still be visualized, but there was vacuolization and variable degrees of myelin disbanding identified. Using serial section transmission electron microscopy, we found that identified synapses could be traced from one section to the next. Using serial block face scanning electron microscopy, we also found that myelinated axons on 2D images can be traced with high fidelity from one image to the next, even at PMIs of up to 27 hours. Collectively, our data corroborate previous findings that immersion fixation is effective for prevention of cellular necrosis and for visualizing many ultrastructural features in at least the surface areas of the brain. However, how structural preservation quality should best be assessed in brain banking is an open question that depends on the intended research applications.
|
| format | Article |
| id | doaj-art-d8a499b225204cb8a9725b245949530b |
| institution | Kabale University |
| issn | 2699-4445 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | University of Münster / Open Journals System |
| record_format | Article |
| series | Free Neuropathology |
| spelling | doaj-art-d8a499b225204cb8a9725b245949530b2025-02-05T07:05:27ZengUniversity of Münster / Open Journals SystemFree Neuropathology2699-44452025-02-01610.17879/freeneuropathology-2025-6104Preservation of cellular structure via immersion fixation in brain bankingMacy Garrood0Emma L. Thorn1Adam Goldstein2Allison Sowa3William Janssen4Alyssa Wilson5Claudia S. López6Raakhee Shankar7Erin S. Stempinski8Kurt Farrell9John F. Crary10Andrew T. McKenzie11Apex Neuroscience, Salem, Oregon, 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, New York, USA Friedman 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, New York, USA Microscopy 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, USADepartments of Neurology and Psychiatry, Icahn School of Medicine at Mount Sinai, New York, New York, USA Multiscale Microscopy Core, Oregon Health and Science University, Portland, Oregon, USA; 7 Department of Biomedical Engineering, Oregon Health and Science University, Portland, Oregon, USAMultiscale Microscopy Core, Oregon Health and Science University, Portland, Oregon, USAMultiscale Microscopy Core, Oregon Health and Science University, Portland, Oregon, 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, New York, USA Friedman 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, New York, USA Apex Neuroscience, Salem, Oregon, USA Immersing the brain in a solution containing formaldehyde is a commonly used method for preserving the structure of human brain tissue in brain banking. However, there are questions about the quality of preservation using this method, as formaldehyde takes a relatively long period of time to penetrate a large organ such as the human brain. As a result, there is a critical need to determine whether immersion fixation is an adequate initial preservation method. To address this, we present exploratory histologic findings from our brain bank following the immersion fixation of hemi-sectioned brain specimens under refrigeration. Using light microscopy, we found that there was no significant change in the size of pericellular or perivascular rarefaction areas based on the postmortem interval (PMI) or on the progression from the outer (frontal cortex) to the inner (striatum) brain regions. Additionally, we did not identify any significant number of ghost cells – a state of late-stage cellular necrosis – in the light micrographs analyzed. Using transmission electron microscopy of tissue from the frontal cortex, we found that synapses could still be visualized, but there was vacuolization and variable degrees of myelin disbanding identified. Using serial section transmission electron microscopy, we found that identified synapses could be traced from one section to the next. Using serial block face scanning electron microscopy, we also found that myelinated axons on 2D images can be traced with high fidelity from one image to the next, even at PMIs of up to 27 hours. Collectively, our data corroborate previous findings that immersion fixation is effective for prevention of cellular necrosis and for visualizing many ultrastructural features in at least the surface areas of the brain. However, how structural preservation quality should best be assessed in brain banking is an open question that depends on the intended research applications. https://unimuenster.de/Ejournals/index.php/fnp/article/view/6104Brain bankingImmersion fixationHistology qualityPostmortem intervalMyelinSynapse |
| spellingShingle | Macy Garrood Emma L. Thorn Adam Goldstein Allison Sowa William Janssen Alyssa Wilson Claudia S. López Raakhee Shankar Erin S. Stempinski Kurt Farrell John F. Crary Andrew T. McKenzie Preservation of cellular structure via immersion fixation in brain banking Free Neuropathology Brain banking Immersion fixation Histology quality Postmortem interval Myelin Synapse |
| title | Preservation of cellular structure via immersion fixation in brain banking |
| title_full | Preservation of cellular structure via immersion fixation in brain banking |
| title_fullStr | Preservation of cellular structure via immersion fixation in brain banking |
| title_full_unstemmed | Preservation of cellular structure via immersion fixation in brain banking |
| title_short | Preservation of cellular structure via immersion fixation in brain banking |
| title_sort | preservation of cellular structure via immersion fixation in brain banking |
| topic | Brain banking Immersion fixation Histology quality Postmortem interval Myelin Synapse |
| url | https://unimuenster.de/Ejournals/index.php/fnp/article/view/6104 |
| work_keys_str_mv | AT macygarrood preservationofcellularstructureviaimmersionfixationinbrainbanking AT emmalthorn preservationofcellularstructureviaimmersionfixationinbrainbanking AT adamgoldstein preservationofcellularstructureviaimmersionfixationinbrainbanking AT allisonsowa preservationofcellularstructureviaimmersionfixationinbrainbanking AT williamjanssen preservationofcellularstructureviaimmersionfixationinbrainbanking AT alyssawilson preservationofcellularstructureviaimmersionfixationinbrainbanking AT claudiaslopez preservationofcellularstructureviaimmersionfixationinbrainbanking AT raakheeshankar preservationofcellularstructureviaimmersionfixationinbrainbanking AT erinsstempinski preservationofcellularstructureviaimmersionfixationinbrainbanking AT kurtfarrell preservationofcellularstructureviaimmersionfixationinbrainbanking AT johnfcrary preservationofcellularstructureviaimmersionfixationinbrainbanking AT andrewtmckenzie preservationofcellularstructureviaimmersionfixationinbrainbanking |