TY - JOUR
T1 - Fast in vitro protocol for the visualization and quantitative high-throughput analysis of sprouting angiogenesis by confocal microscopy
AU - Kempers, Lanette
AU - van der Bijl, Ivo
AU - van Stalborch, Anne-Marieke D
AU - Ponsioen, Bas
AU - Margadant, Coert
N1 - © 2021 The Author(s).
PY - 2021/9/17
Y1 - 2021/9/17
N2 - We describe an optimized, cost-effective, reproducible, and robust protocol to study sprouting angiogenesis in glass-bottom 96-well plates by confocal microscopy, ideal for screening of drug or shRNA libraries. Effective and stable knockdown of gene expression in primary endothelial cells is achieved by lentiviral transduction. Dynamic behavior of individual cells and fluorescent proteins is analyzed by time-lapse imaging, while competitive advantages in tip cell formation are assessed using mixtures of differentially labeled cell populations. Finally, we present a macro for high-throughput analysis. For complete information on the use and execution of this protocol, please refer to van der Bijl et al. (2020) and Kempers et al. (2021).
AB - We describe an optimized, cost-effective, reproducible, and robust protocol to study sprouting angiogenesis in glass-bottom 96-well plates by confocal microscopy, ideal for screening of drug or shRNA libraries. Effective and stable knockdown of gene expression in primary endothelial cells is achieved by lentiviral transduction. Dynamic behavior of individual cells and fluorescent proteins is analyzed by time-lapse imaging, while competitive advantages in tip cell formation are assessed using mixtures of differentially labeled cell populations. Finally, we present a macro for high-throughput analysis. For complete information on the use and execution of this protocol, please refer to van der Bijl et al. (2020) and Kempers et al. (2021).
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85122804641&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/34557696
U2 - https://doi.org/10.1016/j.xpro.2021.100690
DO - https://doi.org/10.1016/j.xpro.2021.100690
M3 - Article
C2 - 34557696
SN - 2666-1667
VL - 2
SP - 100690
JO - STAR Protocols
JF - STAR Protocols
IS - 3
M1 - 100690
ER -