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Glyoxal fixation: An approach to solve immunohistochemical problem in neuroscience research
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- Kohtarou Konno
- Department of Anatomy, Faculty of Medicine, Hokkaido University, Sapporo 060-8638, Japan.
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- Masahiko Watanabe
- Department of Anatomy, Faculty of Medicine, Hokkaido University, Sapporo 060-8638, Japan.
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- Taisuke Miyazaki
- Department of Functioning and Disability, Faculty of Health Sciences, Hokkaido University, Sapporo 060-8638, Japan.
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- Miwako Yamasaki
- Department of Anatomy, Faculty of Medicine, Hokkaido University, Sapporo 060-8638, Japan.
Bibliographic Information
- Published
- 2023-07-14
- Resource Type
- journal article
- DOI
-
- 10.1126/sciadv.adf7084
- Publisher
- American Association for the Advancement of Science (AAAS)
Description
<jats:p>The gold-standard fixative for immunohistochemistry is 4% formaldehyde; however, it limits antibody access to target molecules that are buried within specialized neuronal components, such as ionotropic receptors at the postsynapse and voltage-gated ion channels at the axon initial segment, often requiring additional antigen-exposing techniques to detect their authentic signals. To solve this problem, we used glyoxal, a two-carbon atom di-aldehyde. We found that glyoxal fixation greatly improved antibody penetration and immunoreactivity, uncovering signals for buried molecules by conventional immunohistochemical procedures at light and electron microscopic levels. It also enhanced immunosignals of most other molecules, which are known to be detectable in formaldehyde-fixed sections. Furthermore, we unearthed several specific primary antibodies that were once judged to be unusable in formaldehyde-fixed tissues, allowing us to successfully localize so far controversial synaptic adhesion molecule Neuroligin 1. Thus, glyoxal is a highly effective fixative for immunostaining, and a side-by-side comparison of glyoxal and formaldehyde fixation is recommended for routine immunostaining in neuroscience research.</jats:p>
Journal
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- Science Advances
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Science Advances 9 (28), eadf7084-, 2023-07-14
American Association for the Advancement of Science (AAAS)
