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

Research output: Contribution to journalArticlepeer-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 languageEnglish (US)
Article number102385
JournalSTAR Protocols
Volume4
Issue number3
DOIs
StatePublished - Sep 15 2023

Keywords

  • Cell Biology
  • Developmental Biology
  • Microscopy

ASJC Scopus subject areas

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

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