@inbook{0078f376b3f949ecb7c80d02989a8db8,
title = "Quantifying tissue-specific overexpression of FOXO in Drosophila via mRNA fluorescence in situ hybridization using branched DNA probe technology",
abstract = "While the highly conserved FOXO transcription factors have been studied in Drosophila melanogaster for decades, the ability to accurately control and measure their tissue-specific expression is often cumbersome due to a lack of reagents and to limited, nonhomogeneous samples. The need for quantitation within a distinct cell type is particularly important because transcription factors must be expressed in specific amounts to perform their functions properly. However, the inherent heterogeneity of many samples can make evaluating cell-specific FOXO and/or FOXO load difficult. Here, we describe an extremely sensitive fluorescence in situ hybridization (FISH) approach for visualizing and quantifying multiple mRNAs with single-cell resolution in adult Drosophila cardiomyocytes. The procedure relies upon branched DNA technology, which allows several fluorescent molecules to label an individual transcript, drastically increasing the signal-to-noise ratio compared to other FISH assays. This protocol can be modified for use in various small animal models, tissue types, and for assorted nucleic acids.",
keywords = "Branched DNA, Dorsal vessel, Drosophila melanogaster, FISH, Fluorescence in situ hybridization, Heart tube, RNAscope, ViewRNA, bDNA",
author = "Blice-Baum, {Anna C.} and Georg Vogler and Viswanathan, {Meera C.} and Bosco Trinh and Limpitikul, {Worawan B.} and Anthony Cammarato",
note = "Funding Information: We thank Holly Howarth for excellent technical assistance. This work was supported by R01HL124091 (to A.C.) and Cabrini University Academic Affairs funds (to A.C.B.). For questions, please contact A.C.B. (anna.c.blice-baum@cabrini.edu) or G.V. (gvogler@sbpdiscovery.org). Publisher Copyright: {\textcopyright} 2019, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2019",
doi = "10.1007/978-1-4939-8900-3_15",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "171--190",
booktitle = "Methods in Molecular Biology",
}