Journal
BIOLOGY OPEN
Volume 11, Issue 11, Pages -Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/bio.059417
Keywords
Cell migration; Correlative microscopy; Electron microscopy; Fluorescence microscopy; Focal adhesions
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Focal adhesions (FAs) are important cellular structures that connect the intracellular cytoskeleton to the extracellular matrix. This study used a combination of fluorescence and scanning electron microscopy (SEM) to examine FAs and their associated actin fibres. The results revealed the presence of a highly abundant, novel fork structure at the FA-actin interface in a majority of FAs.
Focal adhesions (FAs) are the main cellular structures to link the intracellular cytoskeleton to the extracellular matrix. FAs mediate cell adhesion, are important for cell migration and are involved in many (patho)-physiological processes. Here we examined FAs and their associated actin fibres using correlative fluorescence and scanning electron microscopy (SEM). We used fluorescence images of cells expressing paxillin-GFP to define the boundaries of FA complexes in SEM images, without using SEM contrast enhancing stains. We observed that SEM contrast was increased around the actin fibre entry site in 98% of FAs, indicating increases in protein density and possibly also phosphorylation levels in this area. In nearly three quarters of the FAs, these nanostructures had a fork shape, with the actin forming the stem and the high-contrast FA areas the fork. In conclusion, the combination of fluorescent and electron microscopy allowed accurate localisation of a highly abundant, novel fork structure at the FA-actin interface.
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