@article{3e60834e645f47519cbc2d4767f51708,
title = "Glyoxal-based fixation of Drosophila embryos for immunofluorescence staining and RNA in situ hybridization",
abstract = "The dialdehyde glyoxal is an alternative chemical fixative that cross-links tissues faster than formaldehyde, retains higher antigenicity, and is less hazardous than either formaldehyde or glutaraldehyde. Here we present a glyoxal-based fixation protocol for use with Drosophila embryos. We describe steps to prepare acid-free glyoxal, fix embryos, and then stain with antibodies for immunofluorescence (IF). We also describe methods for RNA fluorescence in situ hybridization (FISH) and FISH plus IF (FISH-IF) using glyoxal-fixed embryos. This protocol was adapted for Drosophila embryos from the methods of Bussolati et al.1 and Richter et al.2",
keywords = "Cell Biology, Developmental Biology, Microscopy",
author = "Shrunali Amin and Malika Basu and Valeria Buzinova and Anthony Delgado and Tejas Mahadevan and Sanya Mishra and Sarah Zaida and Xi Wang and Sokac, {Anna Marie}",
note = "This work was largely the effort of a team of undergrads working in the Sokac Lab. We wish to thank the more senior lab members who either served as undergrad bench mentors or road-tested and then adopted the method, thus giving us more confidence in its robustness. These senior lab members include Lauren Figard, Natalie Biel, and Poonam Sehgal. We also thank the faculty members who coordinate undergrad research experiences at the students{\textquoteright} respective undergraduate institutions, including Dereth Phillips (Rice University) and Deb Bielser and Stephen Downie (University of Illinois at Urbana Champaign). This work was supported by grants from the National Institutes of Health , to A.M.S. ( R01 GM115111 and R35 GM136384 ). Graphical abstract and Figure 1 were created with BioRender.com .",
year = "2023",
month = sep,
day = "15",
doi = "10.1016/j.xpro.2023.102385",
language = "English (US)",
volume = "4",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "3",
}