Abstract
Lightsheet microscopy is a form of fluorescence microscopy that can be used to visualize specimen with high resolution, a large depth-of-field, and minimal photodamage and photobleaching as compared to traditional confocal microscopy. As this technology becomes much more readily available, it will be useful in revealing new findings in the cardiovascular development field that may be hidden or difficult to image. In this manuscript, we describe an approach for mounting and culturing postimplantation mouse embryos to visualize blood vessel development with a lightsheet microscope.
Key words
- Lightsheet microscopy
- SPIM
- Mouse
- Embryo culture
- Blood vessel
- Angiogenesis
- Vasculogenesis
This is a preview of subscription content, access via your institution.
Buying options
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Drake CJ, Fleming PA (2000) Vasculogenesis in the day 6.5 to 9.5 mouse embryo. Blood 95:1671–1679
Lucitti JL, Jones EA, Huang C, Chen J, Fraser SE, Dickinson ME (2007) Vascular remodeling of the mouse yolk sac requires hemodynamic force. Development 134:3317–3326
Jain RK (2003) Molecular regulation of vessel maturation. Nat Med 9:685–693
Santi PA (2011) Light sheet fluorescence microscopy: a review. J Histochem Cytochem 59:129–138
Huisken J, Swoger J, Del Bene F, Wittbrodt J, Stelzer EH (2004) Optical sectioning deep inside live embryos by selective plane illumination microscopy. Science 305:1007–1009
Winter PW, Shroff H (2014) Faster fluorescence microscopy: advances in high speed biological imaging. Curr Opin Chem Biol 20:46–53
Keller PJ, Schmidt AD, Santella A, Khairy K, Bao Z, Wittbrodt J, Stelzer EH (2010) Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy. Nat Methods 7:637–642
Reeves GT, Trisnadi N, Truong TV, Nahmad M, Katz S, Stathopoulos A (2012) Dorsal-ventral gene expression in the drosophila embryo reflects the dynamics and precision of the dorsal nuclear gradient. Dev Cell 22:544–557
Rieckher M, Kyparissidis-Kokkinidis I, Zacharopoulos A, Kourmoulakis G, Tavernarakis N, Ripoll J, Zacharakis G (2015) A customized light sheet microscope to measure spatio-temporal protein dynamics in small model organisms. PLoS One 10:e0127869
Ichikawa T, Nakazato K, Keller PJ, Kajiura-Kobayashi H, Stelzer EH, Mochizuki A, Nonaka S (2013) Live imaging of whole mouse embryos during gastrulation: migration analyses of epiblast and mesodermal cells. PLoS One 8:e64506
Udan RS, Piazza VG, Hsu CW, Hadjantonakis AK, Dickinson ME (2014) Quantitative imaging of cell dynamics in mouse embryos using light-sheet microscopy. Development 141:4406–4414
Larina IV, Shen W, Kelly OG, Hadjantonakis AK, Baron MH, Dickinson ME (2009) A membrane associated mCherry fluorescent reporter line for studying vascular remodeling and cardiac function during murine embryonic development. Anat Rec 292:333–341
Poche RA, Larina IV, Scott ML, Saik JE, West JL, Dickinson ME (2009) The Flk1-myr::mCherry mouse as a useful reporter to characterize multiple aspects of ocular blood vessel development and disease. Dev Dyn 238:2318–2326
Fraser ST, Hadjantonakis AK, Sahr KE, Willey S, Kelly OG, Jones EA, Dickinson ME, Baron MH (2005) Using a histone yellow fluorescent protein fusion for tagging and tracking endothelial cells in ES cells and mice. Genesis 42:162–171
Dyer MA, Farrington SM, Mohn D, Munday JR, Baron MH (2001) Indian hedgehog activates hematopoiesis and vasculogenesis and can respecify prospective neurectodermal cell fate in the mouse embryo. Development 128:1717–1730
Bambino K, Lacko LA, Hajjar KA, Stuhlmann H (2014) Epidermal growth factor-like domain 7 is a marker of the endothelial lineage and active angiogenesis. Genesis 52:657–670
Jones EA, Crotty D, Kulesa PM, Waters CW, Baron MH, Fraser SE, Dickinson ME (2002) Dynamic in vivo imaging of postimplantation mammalian embryos using whole embryo culture. Genesis 34:228–235
Author information
Authors and Affiliations
Corresponding author
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
Fredrickson, S.J., Hoog, T.G., Udan, R.S. (2021). Visualizing Blood Vessel Development in Cultured Mouse Embryos Using Lightsheet Microscopy. In: Peng, X., Zimmer, W.E. (eds) Cardiovascular Development. Methods in Molecular Biology, vol 2319. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1480-8_11
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1480-8_11
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1479-2
Online ISBN: 978-1-0716-1480-8
eBook Packages: Springer Protocols