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A freeze-substitution approach with solvent-based glyoxal fixative to prevent distortion of ocular structures.
- Source :
- Journal of Histotechnology; Dec2022, Vol. 45 Issue 4, p172-181, 10p
- Publication Year :
- 2022
-
Abstract
- Investigating the function of delicate mammalian eyes often requires chemical fixation, histological sectioning, immunohistochemistry (IHC) and in situ hybridization (ISH). One of the long-standing challenges in the ocular histology field is the limited success of maintaining intact morphology via cryo- or paraffin procedures. Although our latest protocol significantly improved the morphology of mouse eyeball sections, the window technique is time-consuming and requires extensive practice to avoid damage while making windows. In this study, we present a novel glyoxal fixative that is suitable for a freeze-substitution approach to improve both morphology and molecular target preservation of mouse eyes. The method prevents morphology distortion in all tested eyeballs. Therefore, it suits a variety of research needs from morphological examination to investigation of single-molecule RNA expression, using hematoxylin and eosin (H&E) stain, IHC, and ISH assays on either frozen (cryo) or paraffin-infiltrated tissue sections. In addition, this method can be easily performed in many histology laboratories. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01478885
- Volume :
- 45
- Issue :
- 4
- Database :
- Complementary Index
- Journal :
- Journal of Histotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 160402940
- Full Text :
- https://doi.org/10.1080/01478885.2022.2111744