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Method Article
Ceniz Zihni
Matter&Balda Lab, UCL
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Current methodology to quantitatively study microvilli induction by epithelia involves analysis by transmission electron microscopy (TEM) and measurement of microvilli length perpendicular to the membrane. Since this technique involves cutting ultrathin sections of the cell or tissue, the entire surface area of the membrane is not covered and would require a large number of ultrathin sections to reconstruct the surface in order to accurately determine microvilli status. Here we describe a protocol that takes advantage of scanning electron microscopy (SEM) to cover the entire epithelial cell surface of groups of cells and reveals detailed ultrastructure of microvilli. These images can be quantitated using Nikon Imaging Software (NIS) that distinguishes microvilli structures, and even substructures, from flat apical membrane surfaces. The protocol described constitutes a robust assay to quantitatively analyse microvilli generated by cultured epithelia but can potentially also be applied to epithelial tissues and potentially other tissues and cell types that generate microvilli-like structures.
Keywords
epithelia, polarity, differentiation, microvilli
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No conflicting interests
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Associated Publications
An apical MRCK-driven morphogenetic pathway controls epithelial polarity
by Ceniz Zihni, Evi Vlassaks, Stephen Terry, +6
Nature Cell Biology (03 October, 2017)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Ceniz Zihni
Matter&Balda Lab, UCL
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 02 Jan, 2018
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Current methodology to quantitatively study microvilli induction by epithelia involves analysis by transmission electron microscopy (TEM) and measurement of microvilli length perpendicular to the membrane. Since this technique involves cutting ultrathin sections of the cell or tissue, the entire surface area of the membrane is not covered and would require a large number of ultrathin sections to reconstruct the surface in order to accurately determine microvilli status. Here we describe a protocol that takes advantage of scanning electron microscopy (SEM) to cover the entire epithelial cell surface of groups of cells and reveals detailed ultrastructure of microvilli. These images can be quantitated using Nikon Imaging Software (NIS) that distinguishes microvilli structures, and even substructures, from flat apical membrane surfaces. The protocol described constitutes a robust assay to quantitatively analyse microvilli generated by cultured epithelia but can potentially also be applied to epithelial tissues and potentially other tissues and cell types that generate microvilli-like structures.
Figure 1
Figure 2
No conflicting interests
Loading...
Associated Publications
An apical MRCK-driven morphogenetic pathway controls epithelial polarity
by Ceniz Zihni, Evi Vlassaks, Stephen Terry, +6
Nature Cell Biology (03 October, 2017)