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Published on: August 1, 2022
Assessment of murine brain tissue shrinkage caused by different histological fixatives using magnetic resonance and
Hans F Wehrl1, Ilja Bezrukov2, Stefan Wiehr1
1Werner Siemens Imaging Center, Department of Preclinical Imaging and Radiopharmacy, Eberhard Karls University Tuebingen, Tuebingen, Germany.
Abstract:
Especially for neuroscience and the development of new biomarkers, a direct correlation between in vivo imaging and histology is essential. However, this comparison is hampered by deformation and shrinkage of tissue samples caused by fixation, dehydration and paraffin embedding. We used magnetic resonance (MR) imaging and computed tomography (CT) imaging to analyze the degree of shrinkage on murine brains for various fixatives. After in vivo imaging using 7 T MRI, animals were sacrificed and the brains were dissected and immediately placed in different fixatives, respectively: zinc-based fixative, neutral buffered formalin (NBF), paraformaldehyde (PFA), Bouin-Holland fixative and paraformaldehyde-lysine-periodate (PLP). The degree of shrinkage based on mouse brain volumes, radiodensity in Hounsfield units (HU), as well as non-linear deformations were obtained. The highest degree of shrinkage was observed for PLP (68.1%, P < 0.001), followed by PFA (60.2%, P<0.001) and NBF (58.6%, P<0.001). The zinc-based fixative revealed a low shrinkage with only 33.5% (P<0.001). Compared to NBF, the zinc-based fixative shows a slightly higher degree of deformations, but is still more homogenous than PFA. Tissue shrinkage can be monitored non-invasively with CT and MR. Zinc-based fixative causes the smallest degree of brain shrinkage and only small deformations and is therefore recommended for in vivo ex vivo comparison studies.
Insights
A zinc-based fixative minimizes mouse brain shrinkage and deformation during histological processing, enabling better correlation between in vivo imaging and ex vivo analysis for neuroscience research.
Area of Science:
- Neuroscience
- Biomarker Development
- Medical Imaging
Background:
- Accurate correlation between in vivo imaging and histology is crucial for neuroscience and biomarker discovery.
- Tissue processing artifacts, including shrinkage and deformation, hinder direct comparison of imaging and histological data.
Purpose of the Study:
- To quantify and compare the degree of shrinkage and deformation in murine brains using various fixatives after in vivo imaging.
- To identify optimal fixatives for preserving brain tissue integrity for correlative imaging studies.
Main Methods:
- Murine brains underwent in vivo 7 Tesla MRI, followed by dissection and fixation in zinc-based fixative, neutral buffered formalin (NBF), paraformaldehyde (PFA), Bouin-Holland, or paraformaldehyde-lysine-periodate (PLP).
- Computed tomography (CT) and MRI were used to assess brain volume changes, radiodensity (Hounsfield units), and non-linear deformations.
- Statistical analysis was performed to determine the significance of observed shrinkage and deformation differences.
Main Results:
- Paraformaldehyde-lysine-periodate (PLP) caused the highest shrinkage (68.1%), followed by PFA (60.2%) and NBF (58.6%).
- Zinc-based fixative exhibited significantly lower shrinkage (33.5%) and comparable deformation levels to NBF, with better homogeneity than PFA.
- CT and MRI effectively monitored tissue shrinkage non-invasively.
Conclusions:
- Zinc-based fixative is recommended for in vivo-ex vivo comparison studies due to minimal brain shrinkage and controlled deformations.
- Minimizing tissue artifacts is essential for accurate histopathology and biomarker validation in neuroscience research.
