Abstract
Lipid droplets (LDs) are intracellular structures composed of hydrophobic lipids. Their amount in oocytes and embryos varies among the mammalian species and even among different strains of the same species. Here we describe a method to stain LDs, which can be applied to previously fixed mouse oocytes and embryos. This method is based on fluorescent dyes, Nile red and BODIPY, which allow visualization and quantification of LDs using conventional and confocal fluorescence microscopy.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Walther TC, Farese RV Jr (2009) The life of lipid droplets. Biochim Biophys Acta 1791:459–466
Thiam AR, Farese RV Jr, Walther TC (2013) The biophysics and cell biology of lipid droplets. Nat Rev Mol Cell Biol 14:775–786
Valm AM, Cohen S, Legant WR, Melunis J, Hershberg U, Wait E et al (2017) Applying systems-level spectral imaging and analysis to reveal the organelle interactome. Nature 546:162–167
Rambold AS, Cohen S, Lippincott-Schwartz J (2015) Fatty acid trafficking in starved cells: regulation by lipid droplet lipolysis, autophagy and mitochondrial fusion dynamics. Dev Cell 32:678–692
Barbosa AD, Savage DB, Siniossoglou S (2015) Lipid droplet-organelle interactions: emerging roles in lipid metabolism. Curr Opin Cell Biol 35:91–97
Walther TC, Farese RV Jr (2012) Lipid droplets and cellular lipid metabolism. Annu Rev Biochem 81:687–714
McEvoy TG, Coull GD, Broadbent PJ, Hutchinson JSM, Speake BK (2000) Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida. J Reprod Fertil 118:163–170
Musson R, Gasior Ł, Bisogno S, Ptak GE (2022) DNA damage in preimplantation embryos and gametes: specification, clinical relevance and repair strategies. Hum Reprod Update dmab046:1–24
Genicot G, Leroy JLMR, Van SA, Donnay I (2005) The use of a fluorescent dye, Nile red, to evaluate the lipid content of single mammalian oocytes. Theriogenology 63:1181–1194
Yang X, Dunning KR, Wu LL-Y, Hickey TE, Norman RJ, Russell DL et al (2010) Identification of Perilipin-2 as a lipid droplet protein regulated in oocytes during maturation. Reprod Fertil Dev 22:1262–1271
Fam TK, Klymchenko AS, Collot M (2018) Recent advances in fluorescent probes for lipid droplets. Materials (Basel) 11:1768
Bradley J, Pope I, Masia F, Sanusi R, Langbein W, Swann K et al (2016) Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy. Development 143:2238–2247
Held P (2017) Comparison of different cell types for neutral lipid accumulation: using image-based metrics to quantify lipid accumulation in cells. BioTek Instruments- Application note, https://www.biotek.com/assets/tech_resources/NASH%20Cell%20Type%20App%20Note.pdf
Bisogno S, Arena R, Fic K, Gąsior Ł, Ptak G (2020) Lipid droplet utilization by the mouse embryo. Biosci Proc 10:009
Smith R, Wright KL, Ashton L (2016) Raman spectroscopy: an evolving technique for live cell studies. Analyst 141:3590–3600
Perevedentseva E, Krivokharchenko A, Karmenyan AV, Chang HH, Cheng CL (2019) Raman spectroscopy on live mouse early embryo while it continues to develop into blastocyst in vitro. Sci Rep 9:1–12
Di Napoli C, Pope I, Masia F, Watson P, Langbein W, Borri P (2014) Hyperspectral and differential CARS microscopy for quantitative chemical imaging in human adipocytes. Biomed Opt Express 5:1378–1390
Arena R, Bisogno S, Gasior Ł, Rudnicka J, Bernhardt L, Haaf T et al (2021) Lipid droplets in mammalian eggs are utilized during embryonic diapause. Proc Natl Acad Sci U S A 118:e2018362118
Polanski Z, Hoffmann S, Tsurumi C (2005) Oocyte nucleus controls progression through meiotic maturation. Dev Biol 281:184–195
Nagy A, Gertsenstein M, Behringer RR, Vintersten K (2003) Manipulating the mouse embryo: a laboratory manual. NY Cold Spring Harb Lab Press 4:137–146
Acknowledgments
We thank R. Musson for language editing.
Funding
This work was supported by the National Science Centre, Poland (GA no. 2016/21/B/NZ3/03631; GA no. 2019/35/B/NZ4/03547 and 2021/41/B/NZ3/03507 to GEP; and GA no. 2019/35/N/NZ3/03213 to ŁG), and by the European Union’s Horizon 2020 Research and Innovation Programme (GA no. 692185; acronym ERAofART to GEP).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Bisogno, S., Gąsior, Ł., Ptak, G.E. (2023). Nile Red and BODIPY Staining of Lipid Droplets in Mouse Oocytes and Embryos. In: Pellicciari, C., Biggiogera, M., Malatesta, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 2566. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2675-7_16
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2675-7_16
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2674-0
Online ISBN: 978-1-0716-2675-7
eBook Packages: Springer Protocols