Effects of formalin fixation and temperature on MR relaxation times in the human brain
Abstract
Post‐mortem MRI of the brain is increasingly applied in neuroscience for a better understanding of the contrast mechanisms of disease induced tissue changes. However, the influence of chemical processes caused by formalin fixation and differences in temperature may hamper the comparability with results from in vivo MRI. In this study we investigated how formalin fixation and temperature affect T 1 , T 2 and T 2 * relaxation times of brain tissue. Fixation effects were examined with respect to changes in water content and crosslinking. Relaxometry was performed in brain slices from five deceased subjects at different temperatures. All measurements were repeated after 190 days of formaldehyde immersion. The water content of unfixed and fixed tissue was determined using the wet‐to‐dry ratio following drying. Protein weight was determined with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE). Fixation caused a strong decrease of all relaxation times, the strongest effect being seen on T 1 , with a reduction of up to 76%. The temperature coefficient of T 1 was lower in the fixed than unfixed tissue, which was in contrast to T 2 , where an increase of the temperature coefficient was observed following fixation. The reduction of the water content after fixation was in the range of 1–6% and thus not sufficient to explain the changes in relaxation time. Results from SDS‐PAGE indicated a strong increase of the protein size above 260 kDa in all brain structures examined. Our results suggest that crosslinking induced changes of the macromolecular matrix are responsible for T 1 shortening and a decreased temperature dependency. The relaxation times provided in this work should allow optimization of post‐mortem MRI protocols for the brain. Copyright © 2016 John Wiley & Sons, Ltd.
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Fixation can alter tissue microstructure.
“Fixation can alter tissue microstructure47, 48; therefore, NODDI metrics as shown here may not be fully representative of their in vivo counterparts.”
From Neurite dispersion: a new marker of multiple sclerosis spinal cord pathology? (2017) · cites this paper as [48]
Temperature has an effect on MR relaxation times and diffusion.
“As temperature has an effect on MR relaxation times (Birkl et al., 2016) and diffusion (Dhital et al., 2016), tissue temperature during scanning was extracted.”
From Can MRI measure myelin? Systematic review, qualitative assessment, and meta-analysis of studies validating microstructural imaging with myelin histology (2021) · cites this paper as Birkl et al., 2016
Temperature affects MRI parameters such as diffusion and relaxation.
“Another factor to consider is temperature, which affects MRI parameters such as diffusion and relaxation (e.g. Birkl, Langkammer, Golob-Schwarzl, Leoni, Haybaeck, Goessler, Fazekas, Ropele, 2016, Dhital, Labadie, Stallmach, Möller, Turner, 2016).”
From Can MRI measure myelin? Systematic review, qualitative assessment, and meta-analysis of studies validating microstructural imaging with myelin histology (2021) · cites this paper as Birkl, Langkammer, Golob-Schwarzl, Leoni, Haybaeck, Goessler, Fazekas, Ropele, 2016
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Demonstration of Human Fetal Bone Morphology with MR Imaging: A Preliminary Study
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Imaging of the human subthalamic nucleus
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Effects of fixatives on histomagnetic evaluation of iron in rodent spleen
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Deposits of iron oxides in the humanglobus pallidus
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Post-mortem Examination of the Nervous System: Fresh versus Fixed
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Quantification of cortical damage in multiple sclerosis using DTI remains a challenge
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Analytical Dual Flip Angle R1 Calculation Outside the Small‐Angle Regime
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- Authors
- Birkl, C., Langkammer, C., Golob‐Schwarzl, N., Leoni, M., Haybaeck, J., Goessler, W., Fazekas, F., & Ropele, S.
- Affiliations
- Medical University of Graz (AT)
- Harvard University (US)
- University of Graz (AT)
- Corresponding author
- Ropele, Stefan
- ORCID
- Venue
- NMR in Biomedicine
- Publisher
- Wiley
- Published
- 2016-02-02
- Volume
- 29
- Issue
- 4
- Pages
- 458-465
- ISSN
- 0952-3480 (ISSN-L)
- 1099-1492
- OpenAlex
- W2271161903
- Cited by
- 111
- References
- 40
- Citations by year
- 2026: 9 · 2025: 7 · 2024: 13 · 2023: 10 · 2022: 9 · 2021: 23 · 2020: 13 · 2019: 12 · 2018: 8 · 2017: 6 · 2016: 1
- FWCI
- 7.81
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- NMR in Biomedicine (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Oesterreichische Nationalbank (14601)
- Austrian Science Fund (P23576)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Advanced MRI Techniques and Applications
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Advanced Neuroimaging Techniques and Applications
- Physical Sciences › Physics and Astronomy › Nuclear and High Energy Physics › NMR spectroscopy and applications
- Keywords
- formalin fixation, T1 relaxation time, T2 relaxation time, temperature dependence of relaxation, brain water content, protein size, relaxometry, brain slices
- MeSH terms
- Aged, Aged, 80 and over, Brain, Electrophoresis, Polyacrylamide Gel, Female, Formaldehyde, Humans, Magnetic Resonance Imaging, Male, Middle Aged, Temperature, Time Factors, Water, Tissue Fixation
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