Atomic Force Microscopy is a powerful tool for evaluating cell mechanics.
In the recent article “Vertical Light Sheet Enhanced Side-View Imaging for AFM Cell Mechanics Studies” by Kellie Beicker, E. Timothy O’Brien III, Michael R. Falvo, Richard Superfine published in Nature Scientific Reports, the authors combine sideways imaging and a vertical light sheet illumination system integrated with AFM to achieve their results.
5 µm polystyrene beads attached to NanoWorld Arrow-TL1 tipless AFM probes were used.
Figure 5 from Beicker et. al Vertical Light Sheet Enhanced Side-View Imaging for AFM Cell Mechanics Studies: Membrane and nuclear displacements observed in response to force-rupture events between the AFM-tip and cell membrane. (a) Retraction portion of force-indentation curve with important points (A-G) identified. A, the point of zero force application to the cell, B-F, force-rupture peaks, and G, after bead releases from cell. (b) A closer examination of peaks E and F with sub-peaks of the E rupture event identified. No point is shown for E1 because this is the frame immediately following Peak E0. Inset indicates regions where displacement is measured between points E and F highlighted in green. These regions were determined through difference imaging using frames taken at E and F. (c) Regions of cell displacements determined through difference imaging highlighted in green for the sub-peaks indicated in (b). Yellow dashed lines indicate outline of AFM mounted bead. Scale bars = 5 um.
Kellie Beicker, E. Timothy O’Brien III, Michael R. Falvo, Richard Superfine
Vertical Light Sheet Enhanced Side-View Imaging for AFM Cell MechanicsStudies Nature Scientific Reports, volume 8, Article number: 1504 (2018)
DOI: https://doi.org/10.1038/s41598-018-19791-3
The article Beicker et. al, Vertical Light Sheet Enhanced Side-View Imaging for AFM Cell MechanicsStudies is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
We have a month with “R” again and the shellfish season has started in the Northern Hemisphere. So we’d like to share the Nature Communications article by Petrone et. al “Mussel adhesion is dictated by time-regulated secretion and molecular conformation of mussel adhesive proteins” with you.
A NanoWorld Pointprobe®NCSTR AFM probe was used for the AFM images in this paper. This AFM probe is designed to give extra stability and accuracy during soft tapping mode imaging in order to produce higher quality AFM images while minimizing sample damage.
Supplementary Figure 16 from Petrone et. al “Mussel adhesion is dictated by time-regulated secretion and molecular conformation of mussel adhesive proteins”: Atomic Force Microscopy (AFM) of mussel adhesive proteins on mica. AFM images of dry Pvfp-3α and Pvfp-5β adsorbed from 0.02 mg ml-1 solution in 5% acetic acid and 0.25 MO3 on mica. After 20 min adsorption, the mica surfaces were washed with protein -free buffer, and the AFM images show the homogenous distribution of the resulting adsorbed proteins. The height profiles for both proteins are shown in the graphs below, corresponding to the dotted red and blue lines in the respective AFM images (see black arrows).
Luigi Petrone, Akshita Kumar, Clarinda N. Sutanto, Navinkumar J. Patil, Srinivasaraghavan Kannan, Alagappan Palaniappan, Shahrouz Amini, Bruno Zappone, Chandra Verma, Ali Miserez Mussel adhesion is dictated by time-regulated secretion and molecular conformation of mussel adhesive proteins
Nature Communications volume 6, Article number: 8737 (2015)
DOI https://doi.org/10.1038/ncomms9737
The article by Petrone, L.et al. “Mussel adhesion is dictated by time-regulated secretion and molecular conformation of mussel adhesive proteins” is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
For the AFM measurements in the article “Direct observation of the dynamics of single metal ions at the interface with solids in aqueous solutions” by Ricci, M. et al. a NanoWorld Arrow-UHFAuD AFM probe was used. Congratulations to the authors!
Figure 3 from: “Ricci, M. et al. Direct observation of the dynamics of single metal ions at the interface with solids in aqueous solutions.“: Kinetic experiments conducted in pure water (a) show mainly two levels (arrows) when compared to Fig. 2a. Height variations are less pronounced than in RbCl solution and analysis of the surface dynamics (inset) reveals slower timescales with a relatively strong dependence on the choice of threshold. The profile shown in the inset is taken after site averaging (see e.g. Fig. 2d), hence the small height variations. More reliable results were obtained for lower threshold values (here −20 pm, see Supplementary Fig. S10). The overall ratio between the two levels visible in (a) can be changed by adjusting the pH of the water with HCl (b–g), suggesting the higher level to be related to hydration water and the lower level to reflect adsorption of H3O+, as detected by the AFM tip. For each of the pH value studied, the raw kinetic experiments (b,e) are site-averaged (c,f) as in Fig. 2d to remove the mica corrugation and imaging noise. The height distribution of the site-averaged data is then binarised automatically (d,g) depending on whether the surface height is higher or lower than the average between the surface’s highest and lowest points. The fraction of surface interpreted as covered with H3O+ (purple in d and g) changes from 55 ± 3% to 75 ± 2%. (b,e) were acquired with a same tip. The mica samples have been rinsed with the imaging solution after being cleaved and the presence of K+ ions on the surface can be neglected (concentration <10 nM). The scale bar is 3 nm in all experiments.
Abstract:
The dynamics of ions adsorbed at the surface of immersed charged solids plays a central role in countless natural and industrial processes such as crystal growth, heterogeneous catalysis, electrochemistry, or biological function. Electrokinetic measurements typically distinguish between a so-called Stern layer of ions and water molecules directly adsorbed on to the solid’s surface, and a diffuse layer of ions further away from the surface. Dynamics within the Stern layer remain poorly understood, largely owing to a lack of in-situ atomic-level insights. Here we follow the dynamics of single Rb+ and H3O+ ions at the surface of mica in water using high-resolution atomic force microscopy with 25 ms resolution. Our results suggest that single hydrated Rb+ions reside τ1 = 104 ± 5 ms at a given location, but this is dependent on the hydration state of the surface which evolves on a slower timescale of τ2 = 610 ± 30 ms depending on H3O+ adsorption. Increasing the liquid’s temperature from 5 °C to 65 °C predictably decreases the apparent glassiness of the interfacial water, but no clear effect on the ions’ dynamics was observed, indicating a diffusion-dominated process. These timescales are remarkably slow for individual monovalent ions and could have important implications for interfacial processes in electrolytes.
Maria Ricci, William Trewby, Clodomiro Cafolla, Kislon Voïtchovsky Direct observation of the dynamics of single metal ions at the interface with solids in aqueous solutions Nature Scientific Reportsvolume 7, Article number: 43234 (2017)
doi: https://doi.org/10.1038/srep43234
