Induction of Ca2+-Dependent Exocytotic Processes by Laser Ablation of Endothelial Cells

  • Arsila P. K. Ashraf
  • Sophia N. Koerdt
  • Nikita Raj
  • Volker GerkeEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2233)


Ca2+ regulates a variety of cellular processes that are essential to maintain cell integrity and function. Different methods have been used to study these processes by increasing intracellular Ca2+ levels. Here, we describe a protocol to initiate Ca2+-dependent membrane-related events, using laser ablation by near-infrared irradiation. This creates a rupture in the plasma membrane that allows the extracellular Ca2+ to enter the cell and thereby induce a receptor-independent Ca2+ increase. We report laser ablation protocols to study two different Ca2+-induced processes in human endothelial cells—membrane resealing and exocytosis of secretory granules called Weibel-Palade bodies (WPBs). Thus, laser ablation represents a technique that permits the analysis of different Ca2+-regulated processes at high spatiotemporal resolution in a controlled manner.

Key words

Ca2+ Laser ablation Plasma membrane HUVEC FM4-64 WPB exocytosis 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Arsila P. K. Ashraf
    • 1
  • Sophia N. Koerdt
    • 1
  • Nikita Raj
    • 1
  • Volker Gerke
    • 1
    Email author
  1. 1.Institute of Medical Biochemistry, Centre for Molecular Biology of InflammationUniversity of MünsterMünsterGermany

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