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Multimodal super-resolution optical microscopy visualizes the close connection between membrane and the cytoskeleton in liver sinusoidal endothelial cell fenestrations

Permanent lenke
https://hdl.handle.net/10037/8747
DOI
https://doi.org/10.1038/srep16279
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Åpne
article.pdf (1.728Mb)
publisher's pdf (PDF)
Dato
2015
Type
Journal article
Tidsskriftartikkel
Peer reviewed

Forfatter
Mönkemöller, Viola; Øie, Cristina Ionica; Hubner, Wolfgang; Huser, Thomas Rolf; McCourt, Peter Anthony
Sammendrag
Liver sinusoidal endothelial cells (LSECs) act as a filter between blood and the hepatocytes. LSECs are highly fenestrated cells; they contain transcellular pores with diameters between 50 to 200 nm. The small sizes of the fenestrae have so far prohibited any functional analysis with standard and advanced light microscopy techniques. Only the advent of super-resolution optical fluorescence microscopy now permits the recording of such small cellular structures. Here, we demonstrate the complementary use of two different super-resolution optical microscopy modalities, 3D structured illumination microscopy (3D-SIM) and single molecule localization microscopy in a common optical platform to obtain new insights into the association between the cytoskeleton and the plasma membrane that supports the formation of fenestrations. We applied 3D-SIM to multi-color stained LSECs to acquire highly resolved overviews of large sample areas. We then further increased the spatial resolution for imaging fenestrations by single molecule localization microscopy applied to select small locations of interest in the same sample on the same microscope setup. We optimized the use of fluorescent membrane stains for these imaging conditions. The combination of these techniques offers a unique opportunity to significantly improve studies of subcellular ultrastructures such as LSEC fenestrations.
Beskrivelse
Published version. Source at http://doi.org/10.1038/srep16279.
Forlag
Nature Publishing Group
Sitering
Scientific Reports 2015, 5
Metadata
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  • Artikler, rapporter og annet (medisinsk biologi) [1105]

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