From fixed-dried to wet-fixed to live-comparative super-resolution microscopy of liver sinusoidal endothelial cell fenestrations
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https://hdl.handle.net/10037/26411Date
2022-04-20Type
Journal articleTidsskriftartikkel
Peer reviewed
Author
Szafranska, Karolina; Neuman, Tanja; Baster, Zbigniew; Rajfur, Zenon; Szelest, Oskar; Holte, Christopher Florian; Kubisiak, Agata; Kus, Edyta; Wolfson, Deanna; Chlopicki, Stefan; Ahluwalia, Balpreet Singh; Lekka, Malgorzata; Szymonski, Marek; McCourt, Peter Anthony; Zapotoczny, BarlomiejAbstract
Fenestrations in liver sinusoidal endothelial cells
(LSEC) are transcellular nanopores of 50–350 nm diameter
that facilitate bidirectional transport of solutes and macromolecules between the bloodstream and the parenchyma of
the liver. Liver diseases, ageing, and various substances such
as nicotine or ethanol can negatively influence LSECs fenestrations and lead to defenestration. Over the years, the
diameter of fenestrations remained the main challenge for
imaging of LSEC in vitro. Several microscopy, or rather
nanoscopy, approaches have been used to quantify fenestrations in LSEC to assess the effect of drugs and, and toxins in
different biological models. All techniques have their limitations, and measurements of the “true” size of fenestrations
are hampered because of this. In this study, we approach the
comparison of different types of microscopy in a correlative
manner. We combine scanning electron microscopy (SEM)
with optical nanoscopy methods such as structured illumination microscopy (SIM) or stimulated emission depletion
(STED) microscopy. In addition, we combined atomic force
microscopy (AFM) with SEM and STED, all to better understand the previously reported differences between the reports
of fenestration dimensions. We conclude that sample dehydration alters fenestration diameters. Finally, we propose the
combination of AFM with conventional microscopy that allows for easy super-resolution observation of the cell dynamics with additional chemical information that can be
traced back for the whole experiment. Overall, by pairing the
various types of imaging techniques that provide topological
2D/3D/label-free/chemical information we get a deeper
insight into both limitations and strengths of each type microscopy when applied to fenestration analysis.
Publisher
De GruyterCitation
Szafranska, Neuman, Baster, Rajfur, Szelest, Holte, Kubisiak, Kus, Wolfson, Chlopicki, Ahluwalia, Lekka, Szymonski, McCourt, Zapotoczny. From fixed-dried to wet-fixed to live-comparative super-resolution microscopy of liver sinusoidal endothelial cell fenestrations. Nanophotonics. 2022Metadata
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