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

Shrunali Amin; Malika Basu; Valeria Buzinova; Anthony Delgado; Tejas Mahadevan; Sanya Mishra; Sarah Zaida; Xi Wang; Anna Marie Sokac

doi:10.1016/j.xpro.2023.102385

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

Shrunali Amin, Malika Basu, Valeria Buzinova, Anthony Delgado, Tejas Mahadevan, Sanya Mishra, Sarah Zaida, Xi Wang, Anna Marie Sokac

Cell and Developmental Biology

Research output: Contribution to journal › Article › peer-review

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

Original language	English (US)
Article number	102385
Journal	STAR Protocols
Volume	4
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2023.102385
State	Published - Sep 15 2023

Keywords

Cell Biology
Developmental Biology
Microscopy

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

Online availability

10.1016/j.xpro.2023.102385

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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",

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AU - Amin, Shrunali

AU - Basu, Malika

AU - Buzinova, Valeria

AU - Delgado, Anthony

AU - Mahadevan, Tejas

AU - Mishra, Sanya

AU - Zaida, Sarah

AU - Wang, Xi

AU - Sokac, Anna Marie

PY - 2023/9/15

Y1 - 2023/9/15

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AB - 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

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KW - Developmental Biology

KW - Microscopy

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

Abstract

Keywords

ASJC Scopus subject areas

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