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Towards the imaging of Weibel-Palade body biogenesis by serial block
face-scanning electron microscopy
#MMPMID25644989
Mourik MJ
; Faas FGA
; Zimmermann H
; Eikenboom J
; Koster AJ
J Microsc
2015[Aug]; 259
(2
): 97-104
PMID25644989
show ga
Electron microscopy is used in biological research to study the ultrastructure at
high resolution to obtain information on specific cellular processes. Serial
block face-scanning electron microscopy is a relatively novel electron microscopy
imaging technique that allows three-dimensional characterization of the
ultrastructure in both tissues and cells by measuring volumes of thousands of
cubic micrometres yet at nanometre-scale resolution. In the scanning electron
microscope, repeatedly an image is acquired followed by the removal of a thin
layer resin embedded biological material by either a microtome or a focused ion
beam. In this way, each recorded image contains novel structural information
which can be used for three-dimensional analysis. Here, we explore focused ion
beam facilitated serial block face-scanning electron microscopy to study the
endothelial cell-specific storage organelles, the Weibel-Palade bodies, during
their biogenesis at the Golgi apparatus. Weibel-Palade bodies predominantly
contain the coagulation protein Von Willebrand factor which is secreted by the
cell upon vascular damage. Using focused ion beam facilitated serial block
face-scanning electron microscopy we show that the technique has the sensitivity
to clearly reveal subcellular details like mitochondrial cristae and small
vesicles with a diameter of about 50 nm. Also, we reveal numerous associations
between Weibel-Palade bodies and Golgi stacks which became conceivable in
large-scale three-dimensional data. We demonstrate that serial block
face-scanning electron microscopy is a promising tool that offers an alternative
for electron tomography to study subcellular organelle interactions in the
context of a complete cell.
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