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Alkaline phosphatase-based chromogenic and fluorescence detection method for BaseScope™ In Situ hybridization.
- Source :
-
Journal of Histotechnology . Dec2019, Vol. 42 Issue 4, p193-201. 9p. - Publication Year :
- 2019
-
Abstract
- The BaseScope™ assay is a novel, highly sensitive RNA in situ hybridization (ISH) technique, allowing detection of short RNA sequences as well as discrimination between single-nucleotide alterations. Multiplexing BaseScope™ ISH with immunofluorescence assay has proven challenging because the diffusion of colorimetric dyes such as Fast Red in aqueous solutions degrades spatial resolution. In this study, we explore alkaline phosphatase-based fluorescent signal detection methods and integrate it with BaseScope™ RNA ISH. We found that Fast Blue BB/NAMP can be used in BaseScope™ ISH for signal detection. Additionally, we found that the calcium binding fluorochromes calcein and xylenol orange can be used to localize alkaline phosphatase activity in both immunohistochemistry (IHC) and BaseScope™ ISH assays. When applied to mouse brain and intestine tissue sections, we successfully detected circular RNA molecules and cell proliferation antigen Ki-67 proteins. This technological advance expanded the substrate selection of alkaline phosphatase-based BaseScope™ RNA ISH to allow researchers and clinical professionals accurately assess RNA targets with immunofluorescent multiplexing. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01478885
- Volume :
- 42
- Issue :
- 4
- Database :
- Academic Search Index
- Journal :
- Journal of Histotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 141134229
- Full Text :
- https://doi.org/10.1080/01478885.2019.1620906