XB-IMG-171966
Xenbase Image ID: 171966
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Fig. 3. Cell stiffness changes during cell spreading of single cells on fibronectin. (A) Comparing cell stiffness in rounded-up, pre-spread and spread cells by AFM indentation. Cells were explanted onto fibronectin substrates and classified according to their spreading state using an optical side-view setup before elasticity measurements. Cells were considered as having a spread morphology (right image) if the cell diameter at the basal cell side was at least twice the maximal cell height. Furthermore, spread cells formed cellular protrusions on the substrate and displayed no membrane blebbing. Young’s moduli (mean ± SD) were extracted from the AFM indentation force curves by applying a Hertz model fit. Numbers in the data bars denote the number of cells tested per condition. Significance according to a Mann-Whitney-Test, with * = p < 0.05. (B) and (C) Tracking cell spreading and stiffness over time using continuous AFM indentation measurements and time lapse light microscopy. Five single NCCs were seeded side-by-side onto fibronectin and tested continuously by AFM indention in intervals of ∼5 min over several hours. Two still images taken at time 0 min (B) and 240 min (C) from a time lapse series spanning 6 h (contained in Movie S4). Cells initiate cell spreading at different time points. Bar 50 μM. (D) Plotting cell spread area (mean ± SD) and (E) Young’s modulus (mean ± SD) plotted over time for seven representative individual cells measured in experiments exemplified in (B) and (C). Colors indicate same cells in both panels. Linear fits demonstrate an inverse correlation between spread area and stiffness over time for all tested cells. Numbers do not denote same cells in (B), (C) and (D), (E). Image published in: Blaue C et al. (2018) Copyright © 2018. This image is reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View |