Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains
Summary: Visualizing the expression of mRNAs using traditional in situ hybridization is often hampered by obstacles including weak signal, high background, and poor probe specificity. Here, we present a protocol utilizing RNAscope (ACD) to overcome these obstacles and detect multiple types of mRNAs...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725000218 |
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| author | Meilin Wu Vanessa Lambatan Peng Guo William J. Joiner |
| author_facet | Meilin Wu Vanessa Lambatan Peng Guo William J. Joiner |
| author_sort | Meilin Wu |
| collection | DOAJ |
| description | Summary: Visualizing the expression of mRNAs using traditional in situ hybridization is often hampered by obstacles including weak signal, high background, and poor probe specificity. Here, we present a protocol utilizing RNAscope (ACD) to overcome these obstacles and detect multiple types of mRNAs simultaneously in whole-mount adult Drosophila brains. We further describe how mRNAs can be reliably quantified in any cells that can be targeted by common binary expression systems such as Gal4/UAS and labeled by immunohistochemistry.For complete details on the use and execution of this protocol, please refer to De et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-f195becbe6ed4b7b9a8b0c84b29b83bf |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-f195becbe6ed4b7b9a8b0c84b29b83bf2025-02-02T05:29:21ZengElsevierSTAR Protocols2666-16672025-03-0161103615Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brainsMeilin Wu0Vanessa Lambatan1Peng Guo2William J. Joiner3Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USADepartment of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USADepartment of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Nikon Imaging Center, University of California, San Diego, La Jolla, CA 92093, USADepartment of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA; Center for Circadian Biology, University of California, San Diego, La Jolla, CA 92093, USA; Corresponding authorSummary: Visualizing the expression of mRNAs using traditional in situ hybridization is often hampered by obstacles including weak signal, high background, and poor probe specificity. Here, we present a protocol utilizing RNAscope (ACD) to overcome these obstacles and detect multiple types of mRNAs simultaneously in whole-mount adult Drosophila brains. We further describe how mRNAs can be reliably quantified in any cells that can be targeted by common binary expression systems such as Gal4/UAS and labeled by immunohistochemistry.For complete details on the use and execution of this protocol, please refer to De et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725000218single cellmicroscopygene expression |
| spellingShingle | Meilin Wu Vanessa Lambatan Peng Guo William J. Joiner Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains STAR Protocols single cell microscopy gene expression |
| title | Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains |
| title_full | Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains |
| title_fullStr | Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains |
| title_full_unstemmed | Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains |
| title_short | Protocol for multiplexed RNAscope-based imaging of mRNAs in whole-mount adult Drosophila brains |
| title_sort | protocol for multiplexed rnascope based imaging of mrnas in whole mount adult drosophila brains |
| topic | single cell microscopy gene expression |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725000218 |
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