scholarly journals Nanodissected elastically loaded clathrin lattices relax to increased curvature

2021 ◽  
Vol 7 (33) ◽  
pp. eabg9934
Author(s):  
Grigory Tagiltsev ◽  
Christoph A. Haselwandter ◽  
Simon Scheuring
Keyword(s):  
Elastic Energy ◽  
High Speed ◽  
Lattice Relaxation ◽  
Force Microscopy ◽  
Atomic Force ◽  
Afm Imaging ◽  
Constant Area ◽  
Endocytosis Pathway ◽  
Work Supports

Clathrin-mediated endocytosis (CME) is the major endocytosis pathway for the specific internalization of large compounds, growth factors, and receptors. Formation of internalized vesicles from the flat plasma membrane is accompanied by maturation of cytoplasmic clathrin coats. How clathrin coats mature and the mechanistic role of clathrin coats are still largely unknown. Maturation models proposed clathrin coats to mature at constant radius or constant area, driven by molecular actions or elastic energy. Here, combining high-speed atomic force microscopy (HS-AFM) imaging, HS-AFM nanodissection, and elasticity theory, we show that clathrin lattices deviating from the intrinsic curvature of clathrin form elastically loaded assemblies. Upon nanodissection of the clathrin network, the stored elastic energy in these lattices drives lattice relaxation to accommodate an ideal area-curvature ratio toward the formation of closed clathrin-coated vesicles. Our work supports that the release of elastic energy stored in curvature-frustrated clathrin lattices could play a major role in CME.

scholarly journals Quantitative comparison of wideband low-latency phase-locked loop circuit designs for high-speed frequency modulation atomic force microscopy

2018 ◽  
Vol 9 ◽  
pp. 1844-1855 ◽  
Author(s):  
Kazuki Miyata ◽  
Takeshi Fukuma
Keyword(s):  
Atomic Force Microscopy ◽  
Frequency Modulation ◽  
High Speed ◽  
Feedback Loop ◽  
Response Speed ◽  
Low Latency ◽  
Pass Filter ◽  
Force Microscopy ◽  
Atomic Force ◽  
Afm Imaging

A phase-locked loop (PLL) circuit is the central component of frequency modulation atomic force microscopy (FM-AFM). However, its response speed is often insufficient, and limits the FM-AFM imaging speed. To overcome this issue, we propose a PLL design that enables high-speed FM-AFM. We discuss the main problems with the conventional PLL design and their possible solutions. In the conventional design, a low-pass filter with relatively high latency is used in the phase feedback loop, leading to a slow response of the PLL. In the proposed design, a phase detector with a low-latency high-pass filter is located outside the phase feedback loop, while a subtraction-based phase comparator with negligible latency is located inside the loop. This design minimizes the latency within the phase feedback loop and significantly improves the PLL response speed. In addition, we implemented PLLs with the conventional and proposed designs in the same field programmable gate array chip and quantitatively compared their performances. The results demonstrate that the performance of the proposed PLL is superior to that of the conventional PLL: 165 kHz bandwidth and 3.2 μs latency in water. Using this setup, we performed FM-AFM imaging of calcite dissolution in water at 0.5 s/frame with true atomic resolution. The high-speed and high-resolution imaging capabilities of the proposed design will enable a wide range of studies to be conducted on various atomic-scale dynamic phenomena at solid–liquid interfaces.

scholarly journals High‐Speed Atomic Force Microscopy: Large‐Range HS‐AFM Imaging of DNA Self‐Assembly through In Situ Data‐Driven Control (Small Methods 7/2019)

Small Methods ◽  
2019 ◽  
Vol 3 (7) ◽  
pp. 1970022
Author(s):  
Adrian P. Nievergelt ◽  
Christoph Kammer ◽  
Charlène Brillard ◽  
Eva Kurisinkal ◽  
Maartje M. C. Bastings ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Self Assembly ◽  
High Speed ◽  
Large Range ◽  
Data Driven ◽  
Force Microscopy ◽  
In Situ Data ◽  
Atomic Force ◽  
Afm Imaging

A Novel Non-Raster Scan Method for AFM Imaging

2018 ◽  
Author(s):  
Nastaran Nikooienejad ◽  
Mohammad Maroufi ◽  
S. O. Reza Moheimani
Keyword(s):  
High Speed ◽  
High Performance ◽  
Tracking Error ◽  
Contact Mode ◽  
Force Microscopy ◽  
Internal Model Principle ◽  
Atomic Force ◽  
Constant Height ◽  
Afm Imaging ◽  
Raster Scan

We report a new non-raster scan method based on a rosette pattern for high-speed atomic force microscopy (AFM). In this method, the lateral axes of the scanner are driven by the sum of two sinusoids with identical amplitudes and different frequencies. We formulate the problem so as to generate the rosette pattern and calculate scan parameters and resolution. To achieve high performance tracking, a controller is designed based on the internal model principle. The controller includes the dynamic modes of the reference signals and higher harmonics to cope with the system nonlinearities. We conduct an experiment employing the proposed method and a two degree of freedom microelectromechanical system nanopositioner to scan a circular-shaped area with a diameter of 6μm in 0.2 sec. The steady state tracking error is less than 4.48nm, i.e. only 9% of the selected resolution. AFM scanning is performed in contact mode constant height and high quality images are obtained.

scholarly journals Structure and Dynamics of Dinucleosomes Assessed by Atomic Force Microscopy

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Nina A. Filenko ◽  
Dmytro B. Palets ◽  
Yuri L. Lyubchenko
Keyword(s):  
Dynamic Range ◽  
Chromatin Assembly ◽  
Force Microscopy ◽  
Structure And Dynamics ◽  
Atomic Force ◽  
Afm Imaging ◽  
Close Proximity ◽  
Nucleosome Array ◽  
Dna Template

Dynamics of nucleosomes and their interactions are important for understanding the mechanism of chromatin assembly. Internucleosomal interaction is required for the formation of higher-order chromatin structures. Although H1 histone is critically involved in the process of chromatin assembly, direct internucleosomal interactions contribute to this process as well. To characterize the interactions of nucleosomes within the nucleosome array, we designed a dinucleosome and performed direct AFM imaging. The analysis of the AFM data showed dinucleosomes are very dynamic systems, enabling the nucleosomes to move in a broad range along the DNA template. Di-nucleosomes in close proximity were observed, but their population was low. The use of the zwitterionic detergent, CHAPS, increased the dynamic range of the di-nucleosome, facilitating the formation of tight di-nucleosomes. The role of CHAPS and similar natural products in chromatin structure and dynamics is also discussed.

scholarly journals Structural variability and dynamics in the ectodomain of an ancestral-type classical cadherin revealed by AFM imaging

2021 ◽  
Author(s):  
Shigetaka Nishiguchi ◽  
Hiroki Oda
Keyword(s):  
Atomic Force Microscopy ◽  
Structure Function ◽  
High Speed ◽  
Adhesive Properties ◽  
Force Microscopy ◽  
Type Iii ◽  
Atomic Force ◽  
Afm Imaging ◽  
Comparative Structure ◽  
Ancestral Type

Type III cadherin represents the ancestral form of classical cadherin in bilaterian metazoans. Drosophila possesses type III and type IVa cadherins, known as DN- and DE-cadherins, respectively. Mature DN- and DE-cadherins have 15 and 7 extracellular cadherin domain (EC) repeats, respectively, with DN-cadherin EC6–11 homologous to DE-cadherin EC1–6. These EC repeats contain predicted complete or partial Ca2+-free inter-EC linkers that potentially contribute to adhesion. Comparative structure-function studies of DN- and DE-cadherins may help us understand the ancestral and derived states of classical cadherin-mediated adhesion mechanisms. Here, using bead aggregation assays, we found that DN-cadherin EC1–11 and DE-cadherin EC1–6 exhibit Ca2+-dependent adhesive properties. Using high-speed atomic force microscopy (HS-AFM) imaging in solution, we showed that both DN- and DE-cadherin ectodomains share a common morphological framework consisting of a strand-like and a globule-like portion. Furthermore, the DN-cadherin EC repeats were highly variable, flexible in morphology, and with at least three bendable sites, one of which is located in EC6–11 and can act as a flexible hinge. Our findings provide insights into diversification of classical cadherin-mediated adhesion mechanisms. (180 words or less)

scholarly journals Role of trimer–trimer interaction of bacteriorhodopsin studied by optical spectroscopy and high-speed atomic force microscopy

2013 ◽  
Vol 184 (1) ◽  
pp. 2-11 ◽  
Author(s):  
Hayato Yamashita ◽  
Keiichi Inoue ◽  
Mikihiro Shibata ◽  
Takayuki Uchihashi ◽  
Jun Sasaki ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Optical Spectroscopy ◽  
High Speed ◽  
Force Microscopy ◽  
Atomic Force

Brittle Behaviour in Aspirin Crystals: Evidence of Spalling Fracture

2020 ◽  
Author(s):  
Benjamin P. A. Gabriele ◽  
Craig J. Williams ◽  
Douglas Stauffer ◽  
Brian Derby ◽  
Aurora J. Cruz-Cabeza
Keyword(s):  
Surface Roughness ◽  
Atomic Force Microscopy ◽  
Single Crystals ◽  
Spalling Fracture ◽  
Force Microscopy ◽  
Atomic Force ◽  
Afm Imaging

<div> <div> <div> <p>Single crystals of aspirin form I were cleaved and indented on their dominant face. Upon inspection, it was possible to observe strongly anisotropic shallow lateral cracks due to the extreme low surface roughness after cleavage. Atomic Force Microscopy (AFM) imaging showed spalling fractures nucleating from the indent corners, forming terraces with a height of one or two interplanar spacings d100. The formation of such spalling fractures in aspirin was rationalised using basic calculations of attachment energies, showing how (100) layers are poorly bonded when compared to their relatively higher intralayer bonding. An attempt at explaining the preferential propagation of these fractures along the [010] direction is discussed. </p> </div> </div> </div>

Brittle Behaviour in Aspirin Crystals: Evidence of Spalling Fracture

2020 ◽  
Author(s):  
Benjamin P. A. Gabriele ◽  
Craig J. Williams ◽  
Douglas Stauffer ◽  
Brian Derby ◽  
Aurora J. Cruz-Cabeza
Keyword(s):  
Surface Roughness ◽  
Atomic Force Microscopy ◽  
Single Crystals ◽  
Spalling Fracture ◽  
Force Microscopy ◽  
Atomic Force ◽  
Afm Imaging

<div> <div> <div> <p>Single crystals of aspirin form I were cleaved and indented on their dominant face. Upon inspection, it was possible to observe strongly anisotropic shallow lateral cracks due to the extreme low surface roughness after cleavage. Atomic Force Microscopy (AFM) imaging showed spalling fractures nucleating from the indent corners, forming terraces with a height of one or two interplanar spacings d100. The formation of such spalling fractures in aspirin was rationalised using basic calculations of attachment energies, showing how (100) layers are poorly bonded when compared to their relatively higher intralayer bonding. An attempt at explaining the preferential propagation of these fractures along the [010] direction is discussed. </p> </div> </div> </div>

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