Abstract
Immunohistochemistry has been widely used as a robust technique to determine the cellular and subcellular localization of proteins. This information ultimately helps to understand the function of these proteins and how biological processes are regulated. Antibodies applicable for labeling in zebrafish are limited, making immuno-staining challenging. Recently glyoxal fixation was rediscovered in tissue culture, mouse, rat, and Drosophila, expanding the list of effective antibodies for these species. Here, we compare a protocol for zebrafish staining using glyoxal as a fixative agent with PFA. We demonstrate that glyoxal fixation improves the antigenicity of some epitopes thereby increasing the number of useful antibodies in zebrafish.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Han R, Li Z, Fan Y, Jiang Y (2013) Recent advances in super-resolution fluorescence imaging and its applications in biology. J Genet Genomics 40:583–595. https://doi.org/10.1016/j.jgg.2013.11.003
Waldvogel HJ, Curtis MA, Baer K et al (2007) Immunohistochemical staining of post-mortem adult human brain sections. Nat Protoc 1:2719–2732. https://doi.org/10.1038/nprot.2006.354
Inoue D, Wittbrodt J (2011) One for all-a highly efficient and versatile method for fluorescent immunostaining in fish embryos. PLoS One 6:1–7. https://doi.org/10.1371/journal.pone.0019713
Barlow AL, MacLeod A, Noppen S et al (2010) Colocalization analysis in fluorescence micrographs: verification of a more accurate calculation of Pearson’s correlation coefficient. Microsc Microanal 16:710–724. https://doi.org/10.1017/S143192761009389X
Childs GV (2014) History of Immunohistochemistry. Elsevier Inc., Amsterdam
Zhang H, Wen W, Yan J (2017) Application of immunohistochemistry technique in hydrobiological studies. Aquacult Fish 2:140–144. https://doi.org/10.1016/j.aaf.2017.04.004
Perdana (2018) 済無No title. J Chem Inf Model 53:1689–1699. https://doi.org/10.1017/CBO9781107415324.004
Deflorian G, Cinquanta M, Beretta C et al (2009) Monoclonal antibodies isolated by large-scale screening are suitable for labeling adult zebrafish (Danio rerio) tissues and cell structures. J Immunol Methods 346:9–17. https://doi.org/10.1016/j.jim.2009.04.012
Yang J, Xu X (2012) Immunostaining of dissected zebrafish embryonic heart. J Vis Exp 59:e3510. https://doi.org/10.3791/3510
Thomask U (2003) Immunocytochemistry of the amphibian embryo – from overview to ultrastructure. Int J Dev Biol 383:373–383
Moon IS, Cho SJ, Jin IN, Walikonis R (2007) A simple method for combined fluorescence in situ hybridization and immunocytochemistry. Mol Cells 24:76–82
Novak AE, Ribera AB (2003) Immunocytochemistry as a tool for zebrafish developmental neurobiology. Methods Cell Sci 25:79–83. https://doi.org/10.1023/B:MICS.0000006894.43940.b1
Fernández J, Fuentes R (2013) Fixation/permeabilization: new alternative procedure for immunofluorescence and mRNA in situ hybridization of vertebrate and invertebrate embryos. Dev Dyn 242:503–517. https://doi.org/10.1002/dvdy.23943
Richter KN, Revelo NH, Seitz KJ et al (2018) Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy. EMBO J 37:139–159. https://doi.org/10.15252/embj.201695709
Aleström P, D’Angelo L, Midtlyng PJ, et al (2019) Zebrafish: housing and husbandry recommendations. Lab Anim 0:1–12. https://doi.org/10.1177/0023677219869037
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Rostam, N., Dosch, R. (2021). Glyoxal Fixation as an Alternative for Zebrafish Embryo Immunostaining . In: Dosch, R. (eds) Germline Development in the Zebrafish. Methods in Molecular Biology, vol 2218. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0970-5_19
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0970-5_19
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0969-9
Online ISBN: 978-1-0716-0970-5
eBook Packages: Springer Protocols