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Antigenicity is preserved with fixative solutions used in human gross anatomy: A mice brain immunohistochemistry
Eve-Marie Frigon1, Mahsa Dadar2, Denis Boire1
1Department of Anatomy, University of Quebec in Trois-Rivières, Trois-Rivières, QC, Canada.
Insights
Preserving mouse brains with solutions used for gross anatomy dissections maintains tissue quality and antigenicity. This finding supports using human gross anatomy specimens for neuroscientific research and studying brain conditions.
Area of Science:
- Neuroscience
- Histology
- Anatomical Pathology
Background:
- Histology is the gold standard for assessing human brain biology.
- Ex vivo studies utilize brain bank specimens, but gross anatomy labs offer more potential samples.
- The antigenicity preservation of tissues fixed for gross anatomy is not well understood.
Purpose of the Study:
- To evaluate the tissue quality and antigenicity of mouse brains perfused with solutions used for human gross anatomy preservation.
- To compare formaldehyde (FAS), salt-saturated solution (SSS), and alcohol solution (AS) fixatives.
Main Methods:
- Perfusion of 28 C57BL/6J mice brains with FAS (N=9), SSS (N=9), and AS (N=10).
- Sectioning of brains into 40 μm slices for immunohistochemistry.
- Assessment of four key antigens: NeuN (neurons), GFAP (astrocytes), Iba1 (microglia), and PLP (myelin).
Main Results:
- Formaldehyde solution (FAS) yielded better perfusion quality than alcohol solution (AS).
- Salt-saturated solution (SSS) resulted in poor manipulation and lower tissue/immunohistochemistry quality.
- All tested fixatives preserved antigenicity for neuronal nuclei (NeuN), glial fibrillary acidic protein (GFAP), ionized calcium-binding adaptor molecule 1 (Iba1), and myelin proteolipid protein (PLP).
Conclusions:
- Solutions used for gross anatomy specimen preservation maintain brain tissue antigenicity.
- This research validates the potential use of human brains from gross anatomy labs for neuroscientific studies.
- Gross anatomy specimens can be valuable for assessing normal and pathological brain conditions.
Background:
Histology remains the gold-standard to assess human brain biology, so ex vivo studies using tissue from brain banks are standard practice in neuroscientific research. However, a larger number of specimens could be obtained from gross anatomy laboratories. These specimens are fixed with solutions appropriate for dissections, but whether they also preserve brain tissue antigenicity is unclear. Therefore, we perfused mice brains with solutions used for human body preservation to assess and compare the tissue quality and antigenicity of the main cell populations.
Materials And Methods:
Twenty-eight C57BL/6J mice were perfused with 4% formaldehyde (FAS, N = 9), salt-saturated solution (SSS, N = 9), and alcohol solution (AS, N = 10). The brains were cut into 40 μm sections for antigenicity analysis and were assessed by immunohistochemistry of four antigens: neuronal nuclei (NeuN), glial fibrillary acidic protein (GFAP astrocytes), ionized calcium-binding adaptor molecule 1 (Iba1-microglia), and myelin proteolipid protein (PLP). We compared the fixatives according to multiple variables: perfusion quality, ease of manipulation, tissue quality, immunohistochemistry quality, and antigenicity preservation.
Results:
The perfusion quality was better using FAS and worse using AS. The manipulation was very poor in SSS brains. FAS- and AS-fixed brains showed higher tissue and immunohistochemistry quality than the SSS brains. All antigens were readily observed in every specimen, regardless of the fixative solution.
Conclusion:
Solutions designed to preserve specimens for human gross anatomy dissections also preserve tissue antigenicity in different brain cells. This offers opportunities for the use of human brains fixed in gross anatomy laboratories to assess normal or pathological conditions.
