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2018 ; 9
(ä): 379
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Novel Microscopic Techniques for Podocyte Research
#MMPMID30050501
Siegerist F
; Endlich K
; Endlich N
Front Endocrinol (Lausanne)
2018[]; 9
(ä): 379
PMID30050501
show ga
Together with endothelial cells and the glomerular basement membrane, podocytes
form the size-specific filtration barrier of the glomerulus with their
interdigitating foot processes. Since glomerulopathies are associated with
so-called foot process effacement-a severe change of well-formed foot processes
into flat and broadened processes-visualization of the three-dimensional podocyte
morphology is a crucial part for diagnosis of nephrotic diseases. However,
interdigitating podocyte foot processes are too narrow to be resolved by classic
light microscopy due to Ernst Abbe's law making electron microscopy necessary.
Although three dimensional electron microscopy approaches like serial block face
and focused ion beam scanning electron microscopy and electron tomography allow
volumetric reconstruction of podocytes, these techniques are very time-consuming
and too specialized for routine use or screening purposes. During the last few
years, different super-resolution microscopic techniques were developed to
overcome the optical resolution limit enabling new insights into podocyte
morphology. Super-resolution microscopy approaches like three dimensional
structured illumination microscopy (3D-SIM), stimulated emission depletion
microscopy (STED) and localization microscopy [stochastic optical reconstruction
microscopy (STORM), photoactivated localization microscopy (PALM)] reach
resolutions down to 80-20 nm and can be used to image and further quantify
podocyte foot process morphology. Furthermore, in vivo imaging of podocytes is
essential to study the behavior of these cells in situ. Therefore, multiphoton
laser microscopy was a breakthrough for in vivo studies of podocytes in
transgenic animal models like rodents and zebrafish larvae because it allows
imaging structures up to several hundred micrometer in depth within the tissue.
Additionally, along with multiphoton microscopy, lightsheet microscopy is
currently used to visualize larger tissue volumes and therefore image complete
glomeruli in their native tissue context. Alongside plain visualization of
cellular structures, atomic force microscopy has been used to study the change of
mechanical properties of podocytes in diseased states which has been shown to be
a culprit in podocyte maintenance. This review discusses recent advances in the
field of microscopic imaging and demonstrates their currently used and other
possible applications for podocyte research.
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