Revealing The Cell-Material Interface With Nanometer Resolution By FIB-SEM

Santoro, F.; Zhao, W.; Joubert, L.-M.; Duan, L.; Schnitker, J.; van de Burgt, Y.; Lou, H.-Y.; Liu, B.; Salleo, A.; Cui, L.; Cui, Y.; Cui, B.

doi:10.1101/123794

bioRxiv · 10.1101/123794

Revealing The Cell-Material Interface With Nanometer Resolution By FIB-SEM

Santoro, F.·Zhao, W.·Joubert, L.-M.·Duan, L.·Schnitker, J.·van de Burgt, Y.·Lou, H.-Y.·Liu, B.·Salleo, A.·Cui, L.·Cui, Y.·Cui, B.

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Abstract

The interface between biological cells and non-biological surfaces profoundly influences cellular activities, chronic tissue responses, and ultimately the success of medical implants. Materials in contact with cells can be plastics, metal, ceramics or other synthetic materials, and their surfaces vary widely in chemical compositions, stiffness, topography and levels of roughness. To understand the molecular mechanism of how cells and tissues respond to different materials, it is of critical importance to directly visualize the cell-material interface at the relevant length scale of nanometers. Conventional ultrastructural analysis by transmission electron microscopy (TEM) often requires substrate removal before microtome sectioning, which is not only challenging for most substrates but also can cause structural distortions of the interface. Here, we present a new method for in situ examination of the cell-to-material interface at any desired cellular location, based on focused-ion beam milling and scanning electron microscopy imaging (FIB-SEM). This method involves a thin-layer plastification procedure that preserves adherent cells as well as enhances the contrast of biological specimen. We demonstrate that this unique procedure allows the visualization of cell-to-material interface and intracellular structures with 10nm resolution, compatible with a variety of materials and surface topographies, and capable of volume and multi-directional imaging. We expect that this method will be very useful for studies of cell-to-material interactions and also suitable for in vivo studies such as examining osteoblast adhesion and new bone formation in response to titanium implants.

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Santoro, F., Zhao, W., Joubert, L.-M., Duan, L., Schnitker, J., van de Burgt, Y., Lou, H.-Y., Liu, B., Salleo, A., Cui, L., Cui, Y., Cui, B.. 2017-04-04. Revealing The Cell-Material Interface With Nanometer Resolution By FIB-SEM. https://doi.org/10.1101/123794

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