Glyoxal-based fixation of Drosophila embryos for immunofluorescence staining and RNA in situ hybridization.

Autor:	Amin S; University of Illinois at Urbana Champaign, Department of Cell & Developmental Biology, Urbana, IL 61801, USA; University of Illinois at Urbana Champaign, School of Molecular & Cellular Biology, Urbana, IL 61801, USA. Electronic address: sjamin2@illinois.edu., Basu M; University of Illinois at Urbana Champaign, School of Integrative Biology, Urbana, IL 61801, USA; Johns Hopkins University, Department of Molecular Microbiology & Immunology, Baltimore, MD 21205, USA., Buzinova V; University of Illinois at Urbana Champaign, School of Molecular & Cellular Biology, Urbana, IL 61801, USA; University of Kentucky, Department of Molecular & Cellular Biochemistry, Lexington, KY 40536, USA., Delgado A; University of Illinois at Urbana Champaign, School of Molecular & Cellular Biology, Urbana, IL 61801, USA; University of Illinois at Chicago, College of Medicine, Chicago, IL 60612, USA., Mahadevan T; University of Illinois at Urbana Champaign, Department of Cell & Developmental Biology, Urbana, IL 61801, USA; University of Illinois at Urbana Champaign, School of Molecular & Cellular Biology, Urbana, IL 61801, USA., Mishra S; University of Illinois at Urbana Champaign, School of Molecular & Cellular Biology, Urbana, IL 61801, USA; University of Texas Health Science Center, School of Biomedical Sciences, Katy, TX 77494, USA., Zaida S; University of Illinois at Urbana Champaign, School of Molecular & Cellular Biology, Urbana, IL 61801, USA; Northwestern Medicine, Chicago, IL 60605, USA., Wang X; Department of BioSciences, Rice University, Houston, TX 77251, USA; Baylor College of Medicine, Department of Biochemistry, Houston, TX 77030, USA; University of Mississippi Medical Center, Department of Pharmacology & Toxicology, Jackson, MS 39216, USA., Sokac AM; University of Illinois at Urbana Champaign, Department of Cell & Developmental Biology, Urbana, IL 61801, USA; University of Illinois at Urbana Champaign, School of Molecular & Cellular Biology, Urbana, IL 61801, USA; Baylor College of Medicine, Department of Biochemistry, Houston, TX 77030, USA. Electronic address: asokac@illinois.edu.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2023 Sep 15; Vol. 4 (3), pp. 102385. Date of Electronic Publication: 2023 Jul 04.
DOI:	10.1016/j.xpro.2023.102385
Abstrakt:	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 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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