Imaging fenestrations in liver sinusoidal endothelial cells by optical localization microscopy

Viola Mönkemöller, Mark Schüttpelz, Peter McCourt, Karen Sørensen, Bård Smedsrød, Thomas R Huser

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18 Scopus citations

Abstract

Liver sinusoidal endothelial cells (LSEC) are an important class of endothelial cells facilitating the translocation of lipoproteins and small molecules between the liver and blood. A number of clinical conditions, especially metabolic and aging-related disorders, are implicated by improper function of LSECs. Despite their importance, research into these cells is limited because the primary ultrastructures involved in their function are transcellular pores, called fenestrations, with diameters in a size range between 50-200 nm, i.e. well below the optical diffraction limit. Here, we show that we are able to resolve fenestrations with a spatial resolution of ∼20 nm by direct stochastic optical reconstruction microscopy (dSTORM). The cellular plasma membrane was labeled at high fluorophore density with CellMask Deep Red and imaged using a reducing buffer system. We compare the higher degree of structural detail that dSTORM provides to results obtained by 3D structured illumination microscopy (3D-SIM). Our results open up a path to image these physiologically important cells in vitro using highly resolving localization microscopy techniques that could be implemented on non-specialized fluorescence microscopes, enabling their investigation in most biomedical laboratories without the need for electron microscopy.

Original languageEnglish (US)
Pages (from-to)12576-12581
Number of pages6
JournalPhysical Chemistry Chemical Physics
Volume16
Issue number24
DOIs
StatePublished - Jun 28 2014

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ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Physics and Astronomy(all)

Cite this

Mönkemöller, V., Schüttpelz, M., McCourt, P., Sørensen, K., Smedsrød, B., & Huser, T. R. (2014). Imaging fenestrations in liver sinusoidal endothelial cells by optical localization microscopy. Physical Chemistry Chemical Physics, 16(24), 12576-12581. https://doi.org/10.1039/c4cp01574f