| Is optimal cutting temperature compound essential embedding solution treatment to cryo-sectioning of brain tissue? |
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Hye Kyung Baek, Ji Ae Song, Sun Shin Yi |
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Department of Biomedical Laboratory Science, College of Medical Sciences, Soonchunhyang University |
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| Abstract |
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We tested a set of conditions for obtaining optimal tissue quality in preparation for histology in samples of mouse brain. C57BL/6J mice were sacrificed and perfused with 4% paraformaldehyde, after which the brains were removed and dehydrated in 30% sucrose solution. The brains were then divided into four groups according to freezing temperature and usage of optimal cutting temperature (OCT) compound. Next, we stained the sectioned brain tissues with Harris hematoxylin and eosin Y and immunohistochemistry was performed for doublecortin. The best quality tissue was obtained at $-25^{circ}C$ and by not embedding with the OCT compound. When frozen at $-25^{circ}C$, the embedded tissue was significantly damaged by crystals, while at $-80^{circ}C$ there were no meaningful differences between qualities of embedded- and non-embedded tissues. Overall, we identified a set of conditions to obtain quality frozen brain sections. Our developed protocol will help resolve matters associated with damage caused to sectioned brain tissue by crystal formation during freezing. |
| Key Words:
brain tissue, cryosection, freezing temperature, optimal cutting temperature compound, tissue quality |
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