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Analysis of Fluid-Phase Endocytosis in (Intact) Plant Cells

  • Vera Bandmann
  • Peter Haub
  • Tobias MeckelEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1209)

Abstract

Endocytosis is a continuous process at the plasma membrane at least of all eukaryotic cells. Regardless of the molecular machinery, which drives the formation and uptake of endocytic vesicles, it is reasonable to assume that this process inevitably collects external fluid. Hence, at least for the majority of apoplastic solutes, the endocytosis of the fluid phase is likely to be an inevitable process. Due to its independence from the molecular machinery and low selectivity with respect to the cargo, it is thus perfectly suited to be used as a tracer to follow the activity of all endocytic events. Here we describe simple protocols based on fluorescence microscopy, which yield quantitative information about endocytic vesicle sizes—with sub-diffraction accuracy—as well as the size exclusion limits for these uptake routes.

Key words

Fluid-phase endocytosis Fluorescence Microscopy Confocal Single-molecule Diffraction limit Fluorescence intensity Size exclusion limit 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Plant Cell Biology, Department of BiologyINM-Leibniz-Institute for New Materials66123 SaarbrückenGermany
  2. 2.DIPsystems.deAltlussheimGermany
  3. 3.Membrane Dynamics, Department of BiologyTechnische Universität Darmstadt65287 DarmstadtGermany

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