Tag: biology AFM probes

Carbon nanotube porin diffusion in mixed composition supported lipid bilayers

Lipid membranes play a key role in living systems by providing a structural barrier that separates cellular compartments. Bilayer fluidity in the lateral plane is a key property of lipid membranes, that allows the membrane to have sufficient flexibility to accommodate dynamic stresses, shape changes and rearrangements accompanying the cellular lifecycle.*

In the article “Carbon nanotube porin diffusion in mixed composition supported lipid bilayers” Kylee Sullivan, Yuliang Zhang, Joseph Lopez, Mary Lowe and Aleksandr Noy describe how they used high-speed atomic force microscopy (HS-AFM) and all-atom molecular dynamics (MD) simulations to study the behavior of CNTPs in a mixed lipid membrane consisting of DOPC lipid with a variable percentage of DMPC lipid added to it. HS-AFM data reveal that the CNTPs undergo diffusive motion in the bilayer plane.*

Motion trajectories extracted from the HS-AFM movies indicate that CNTPs exhibit diffusion coefficient values broadly similar to values reported for membrane proteins in supported lipid bilayers. The data also indicate that increasing the percentage of DMPC leads to a marked slowing of CNTP diffusion. MD simulations reveal a CNTP-lipid assembly that diffuses in the membrane and show trends that are consistent with the experimental observations. *

The above-mentioned study confirms that CNTPs mimic the major features of the diffusive movement of biological pores in lipid membranes and shows how the increase in bilayer viscosity leads to a corresponding slowdown in protein motion. It should be possible to extend this approach to studies of other membrane protein dynamics in supported lipid bilayers. The authors note that those studies, however, will need to be mindful of the challenge of unambiguous visualization of the membrane components, especially in systems that incorporate smaller proteins, such as antimicrobial peptides. Another challenge that could complicate these studies would be microscopic phase separation of the lipid matrix that could lead to complicated pore dynamics in the membrane. *

NanoWorld Ultra-Short AFM cantilevers with high-density carbon/diamond-like carbon (HDC/DLC) AFM tips of the USC-F1.2-k0.15 type were used for the high-speed atomic force microscopy described in the article. *

Figure 2 from “Carbon nanotube porin diffusion in mixed composition supported lipid bilayers” by Kylee Sullivan et al.: CNTP motion in supported lipid bilayers. (a) Representative frames (with times in seconds indicated on each image) from an HS-AFM movie showing a CNTP diffusing in a supported lipid bilayer with 80:20 DOPC-DMPC ratio (see also Supplementary Movie 2). (b) A representative trajectory for CNTP diffusion in the bilayer. The time step between each datapoint is 0.5 s. NanoWorld Ultra-Short AFM Cantilvers USC-F1.2-k0.15 were used for the HS-AFM imaging
Figure 2 a and b from “Carbon nanotube porin diffusion in mixed composition supported lipid bilayers” by Kylee Sullivan et al.:

CNTP motion in supported lipid bilayers. (a) Representative frames (with times in seconds indicated on each image) from an HS-AFM movie showing a CNTP diffusing in a supported lipid bilayer with 80:20 DOPC-DMPC ratio (see also Supplementary Movie 2). (b) A representative trajectory for CNTP diffusion in the bilayer. The time step between each datapoint is 0.5 s.
Please refer to the full article cited below for the full figure.

*Kylee Sullivan, Yuliang Zhang, Joseph Lopez, Mary Lowe and Aleksandr Noy
Carbon nanotube porin diffusion in mixed composition supported lipid bilayers
Nature Scientific Reports volume 10, Article number: 11908 (2020)
DOI: https://doi.org/10.1038/s41598-020-68059-2

Please follow this external link to read the full article: https://rdcu.be/b69wj

Open Access : The article “Carbon nanotube porin diffusion in mixed composition supported lipid bilayers” by Kylee Sullivan, Yuliang Zhang, Joseph Lopez, Mary Lowe and Aleksandr Noy 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 https://creativecommons.org/licenses/by/4.0/.

NanoWorld Screencast on Ultra-Short Cantilevers (USC) for High-Speed AFM (HS-AFM) and video rate Atomic Force Microscopy passes 1000 views mark

The screencast on Ultra-Short Cantilevers (USC) for High Speed AFM (HS-AFM)  and video rate Atomic Force Microscopy held by Mathieu Burri  has just reached the 1000 views mark. Congratulations Mathieu!

The Ultra-Short Cantilevers series combines very small cantilevers capable of resonating in the MHz regime and a very sharp and wear resistant tip and is dedicated to High-Speed AFM (HS-AFM). The Ultra-Short Cantilevers series consists of six different types of probes which cover the complete range of high speed scanning applications, from non-contact mode to contact mode, from measurements in air to measurements in liquid.

More information such as high speed videos, an image gallery and a regularly updated reference list of scientific articles on high speed AFM can be found on the dedicated website: https://www.highspeedscanning.com/ . You can also find references to scientific articles mentioning the use of NanoWorld USC AFM probes on the NanoWorld blog.

If you haven’t seen the USC screencast yet have a look at:

NanoWorld ULTRA-SHORT CANTILEVERS for High-Speed AFM

A Chinese Version of this screencast is also available:

Human ESCRT-III polymers assemble on positively curved membranes and induce helical membrane tube formation

The Endosomal Sorting Complex Required for Transport-III (ESCRT-III) is part of a conserved membrane remodeling machine. ESCRT-III employs polymer formation to catalyze inside-out membrane fission processes in a large variety of cellular processes, including budding of endosomal vesicles and enveloped viruses, cytokinesis, nuclear envelope reformation, plasma membrane repair, exosome formation, neuron pruning, dendritic spine maintenance, and preperoxisomal vesicle biogenesis.*

How membrane shape influences ESCRT-III polymerization and how ESCRT-III shapes membranes is yet unclear.*

In the article “Human ESCRT-III polymers assemble on positively curved membranes and induce helical membrane tube formation” Aurélie Bertin, Nicola de Franceschi, Eugenio de la Mora, Sourav Maity, Maryam Alqabandi, Nolwen Miguet, Aurélie di Cicco, Wouter H. Roos, Stéphanie Mangenot, Winfried Weissenhorn and Patricia Bassereau describe how human core ESCRT-III proteins, CHMP4B, CHMP2A, CHMP2B and CHMP3 are used to address this issue in vitro by combining membrane nanotube pulling experiments, cryo-electron tomography and Atomic Force Microscopy.*

The authors show that CHMP4B filaments preferentially bind to flat membranes or to tubes with positive mean curvature.*

The results presented in the article cited above underline the versatile membrane remodeling activity of ESCRT-III that may be a general feature required for cellular membrane remodeling processes.*

The authors provide novel insight on how mechanics and geometry of the membrane and of ESCRT-III assemblies can generate forces to shape a membrane neck.*

NanoWorld Ultra-Short AFM Cantilevers USC-F1.2-k0.15 were used for the High-speed Atomic Force Microscopy ( HS-AFM ) experiments presented in this article.*

Figure 1 from «Human ESCRT-III polymers assemble on positively curved membranes and induce helical membrane tube formation” by Aurélie Bertin et al.: CHMP4-ΔC flattens LUVs and binds preferentially to flat membranes or to membranes with a positive mean curvature. 1a CHMP4B-ΔC spirals observed by HS-AFM on a lipid bilayer. Scale bar: 50 nm. Please refer to the full article for the complete figure: https://rdcu.be/b5rOe
Figure 1 from «Human ESCRT-III polymers assemble on positively curved membranes and induce helical membrane tube formation” by Aurélie Bertin et al.:
CHMP4-ΔC flattens LUVs and binds preferentially to flat membranes or to membranes with a positive mean curvature.
1a CHMP4B-ΔC spirals observed by HS-AFM on a lipid bilayer. Scale bar: 50 nm.
Please refer to the full article for the complete figure: https://rdcu.be/b5rOe

*Aurélie Bertin, Nicola de Franceschi, Eugenio de la Mora, Sourav Maity, Maryam Alqabandi, Nolwen Miguet, Aurélie di Cicco, Wouter H. Roos, Stéphanie Mangenot, Winfried Weissenhorn and Patricia Bassereau
Human ESCRT-III polymers assemble on positively curved membranes and induce helical membrane tube formation
Nature Communications volume 11, Article number: 2663 (2020)
DOI: https://doi.org/10.1038/s41467-020-16368-5

Please follow this external link to read the full article: https://rdcu.be/b5rOe

Open Access The article “ Human ESCRT-III polymers assemble on positively curved membranes and induce helical membrane tube formation “ by Aurélie Bertin, Nicola de Franceschi, Eugenio de la Mora, Sourav Maity, Maryam Alqabandi, Nolwen Miguet, Aurélie di Cicco, Wouter H. Roos, Stéphanie Mangenot, Winfried Weissenhorn and Patricia Bassereau 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/.