PMID- 37405926
OWN - NLM
STAT- MEDLINE
DCOM- 20240308
LR  - 20240308
IS  - 2666-1667 (Electronic)
IS  - 2666-1667 (Linking)
VI  - 4
IP  - 3
DP  - 2023 Sep 15
TI  - Glyoxal-based fixation of Drosophila embryos for immunofluorescence staining and 
      RNA in situ hybridization.
PG  - 102385
LID - S2666-1667(23)00352-0 [pii]
LID - 10.1016/j.xpro.2023.102385 [doi]
LID - 102385
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).
CI  - Copyright (c) 2023 The Author(s). Published by Elsevier Inc. All rights reserved.
FAU - Amin, Shrunali
AU  - Amin S
AD  - 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.
FAU - Basu, Malika
AU  - Basu M
AD  - 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.
FAU - Buzinova, Valeria
AU  - Buzinova V
AD  - 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.
FAU - Delgado, Anthony
AU  - Delgado A
AD  - 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.
FAU - Mahadevan, Tejas
AU  - Mahadevan T
AD  - 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.
FAU - Mishra, Sanya
AU  - Mishra S
AD  - 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.
FAU - Zaida, Sarah
AU  - Zaida S
AD  - University of Illinois at Urbana Champaign, School of Molecular & Cellular 
      Biology, Urbana, IL 61801, USA; Northwestern Medicine, Chicago, IL 60605, USA.
FAU - Wang, Xi
AU  - Wang X
AD  - 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.
FAU - Sokac, Anna Marie
AU  - Sokac AM
AD  - 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.
LA  - eng
GR  - R01 GM115111/GM/NIGMS NIH HHS/United States
GR  - R35 GM136384/GM/NIGMS NIH HHS/United States
PT  - Journal Article
DEP - 20230704
PL  - United States
TA  - STAR Protoc
JT  - STAR protocols
JID - 101769501
RN  - 63231-63-0 (RNA)
RN  - 50NP6JJ975 (Glyoxal)
RN  - 1HG84L3525 (Formaldehyde)
SB  - IM
MH  - Animals
MH  - *Drosophila/genetics
MH  - *RNA
MH  - Tissue Fixation/methods
MH  - Glyoxal
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Formaldehyde
MH  - Fluorescent Antibody Technique
PMC - PMC10345161
OTO - NOTNLM
OT  - Cell Biology
OT  - Developmental Biology
OT  - Microscopy
COIS- Declaration of interests The authors declare no competing interests.
EDAT- 2023/07/05 19:12
MHDA- 2024/03/08 06:43
PMCR- 2023/07/04
CRDT- 2023/07/05 12:36
PHST- 2023/02/24 00:00 [received]
PHST- 2023/05/25 00:00 [revised]
PHST- 2023/05/25 00:00 [accepted]
PHST- 2024/03/08 06:43 [medline]
PHST- 2023/07/05 19:12 [pubmed]
PHST- 2023/07/05 12:36 [entrez]
PHST- 2023/07/04 00:00 [pmc-release]
AID - S2666-1667(23)00352-0 [pii]
AID - 102385 [pii]
AID - 10.1016/j.xpro.2023.102385 [doi]
PST - ppublish
SO  - STAR Protoc. 2023 Sep 15;4(3):102385. doi: 10.1016/j.xpro.2023.102385. Epub 2023 
      Jul 4.