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A novel ex vivo, in situ method to study the human brain through MRI and histology
Josefina Maranzano1, Mahsa Dadar2, Antony Bertrand-Grenier3
1Department of Anatomy, Université du Québec à Trois-Rivières (UQTR), Trois-Rivières, Québec, Canada(2); McConnell Brain Imaging Centre, Montreal Neurological Institute, McGill University, Montréal, Québec, Canada.
Background:
MRI-histology correlation studies of the ex vivo brain mostly employ fresh, extracted (ex situ) specimens, aldehyde fixed by immersion, which has several disadvantages for MRI scanning (e.g. deformation of the organ). A minority of studies are done ex vivo-in situ (unfixed brain), requiring an MRI scanner readily available within a few hours of the time of death.
New Method:
We propose a new technique, exploited by anatomists, for scanning the ex vivo brain: fixation by whole body perfusion, which implies fixation of the brain in situ. This allows scanning the brain surrounded by fluids, meninges, and skull, preserving the structural relationships of the brain in vivo. To evaluate the proposed method, five heads perfused-fixed with a saturated sodium chloride solution were employed. Three sequences were acquired on a 1.5 T MRI scanner: T1weighted, T2weighted-FLAIR, and Gradient-echo. Histology analysis included immunofluorescence for myelin basic protein and neuronal nuclei.
Results:
All MRIs were successfully processed through a validated pipeline used with in vivo MRIs. All cases exhibited positive antigenicity for myelin and neuronal nuclei.
Comparison With Existing Methods:
All scans registered to a standard neuroanatomical template in pseudo-Talairach space more accurately than an ex vivo-ex situ scan. The time interval to scan the ex vivo brain in situ was increased to at least 10 months.
Conclusions:
MRI and histology study of the ex vivo-in situ brain fixed by perfusion is an alternative approach that has important procedural and practical advantages over the two standard methods to study the ex vivo brain.
Insights
Whole body perfusion fixation enables ex vivo brain MRI scanning in situ, preserving in vivo structural relationships. This novel method offers advantages over traditional ex situ techniques for brain imaging and histology correlation.
Area of Science:
- Neuroimaging
- Anatomical Pathology
- Medical Imaging Techniques
Background:
- Ex vivo brain MRI studies commonly use fresh or immersion-fixed specimens, leading to deformation and limited scanning windows.
- Current methods for ex vivo brain imaging include ex situ (fresh) or ex vivo-in situ (unfixed) approaches, each with significant limitations.
Purpose of the Study:
- To introduce and evaluate a novel technique for ex vivo brain MRI scanning using whole-body perfusion fixation for in situ preservation.
- To assess the feasibility and advantages of perfusion fixation for maintaining brain structure and enabling advanced MRI analysis.
Main Methods:
- Whole body perfusion fixation with saturated sodium chloride solution was employed for ex vivo brain specimens.
- Magnetic Resonance Imaging (MRI) was performed using T1-weighted, T2-weighted-FLAIR, and Gradient-echo sequences on a 1.5T scanner.
- Histology analysis included immunofluorescence for myelin basic protein and neuronal nuclei.
Main Results:
- All MRI scans were successfully processed using a validated pipeline, comparable to in vivo data.
- Perfusion-fixed ex vivo brains demonstrated positive antigenicity for myelin and neuronal nuclei.
- Scans registered more accurately to a neuroanatomical template compared to ex situ specimens, with extended scanning viability up to 10 months.
Conclusions:
- Whole body perfusion fixation offers a superior alternative for ex vivo brain MRI and histology correlation.
- This in situ fixation method preserves crucial structural relationships and overcomes limitations of traditional ex vivo techniques.
- The technique provides procedural and practical advantages for studying the ex vivo brain.
