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

Scientific reports, 2015-11, Vol.5 (1), p.16279-16279, Article 16279 [Peer Reviewed Journal]

Copyright Nature Publishing Group Nov 2015 ;info:eu-repo/semantics/openAccess ;Copyright © 2015, Macmillan Publishers Limited 2015 Macmillan Publishers Limited ;ISSN: 2045-2322 ;EISSN: 2045-2322 ;DOI: 10.1038/srep16279 ;PMID: 26549018

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Title:
Multimodal super-resolution optical microscopy visualizes the close connection between membrane and the cytoskeleton in liver sinusoidal endothelial cell fenestrations
Author: Mönkemöller, Viola ; Øie, Cristina ; Hübner, Wolfgang ; Huser, Thomas ; McCourt, Peter
Subjects: Actin ; Animals ; Applied optics ; Basale biofag: 470 ; Basic biosciences: 470 ; Biofysikk: 477 ; Biophysics: 477 ; Cell Membrane - ultrastructure ; Color ; Cytoskeleton ; Cytoskeleton - ultrastructure ; Endothelial cells ; Endothelial Cells - ultrastructure ; Hepatocytes ; Hepatocytes - ultrastructure ; Illumination ; Liver - ultrastructure ; Localization ; Matematikk og Naturvitenskap: 400 ; Mathematics and natural science: 400 ; Microscopes ; Microscopy ; Microscopy, Fluorescence ; Molecular biophysics ; Multimodal Imaging ; Rats ; Spatial discrimination ; Super-resolution microscopy ; VDP
Is Part Of: Scientific reports, 2015-11, Vol.5 (1), p.16279-16279, Article 16279
Description: 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.
Publisher: England: Nature Publishing Group
Language: English
Identifier: ISSN: 2045-2322
EISSN: 2045-2322
DOI: 10.1038/srep16279
PMID: 26549018
Source: MEDLINE
NORA Norwegian Open Research Archives
PubMed Central
ProQuest Central
DOAJ Directory of Open Access Journals

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

Copyright Nature Publishing Group Nov 2015 ;info:eu-repo/semantics/openAccess ;Copyright © 2015, Macmillan Publishers Limited 2015 Macmillan Publishers Limited ;ISSN: 2045-2322 ;EISSN: 2045-2322 ;DOI: 10.1038/srep16279 ;PMID: 26549018

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