Atomic Force Microscopy of Biological Samples

MRS Bulletin ◽  
2004 ◽  
Vol 29 (7) ◽  
pp. 449-455 ◽  
Author(s):  
P.L.T.M. Frederix ◽  
B.W. Hoogenboom ◽  
D. Fotiadis ◽  
D.J. Müller ◽  
A. Engel
Keyword(s):  
Mechanical Stability ◽  
Signal To Noise Ratio ◽  
Single Molecules ◽  
Scanning Speed ◽  
Surface Dynamics ◽  
Buffer Solution ◽  
Force Extension ◽  
Force Microscopy ◽  
Force Sensitivity ◽  
Atomic Force

AbstractThe atomic force microscope (AFM) allows biomolecules to be observed and manipulated under native conditions. It produces images with an outstanding signal-to-noise ratio and addresses single molecules while the sample is in a buffer solution. Progress in sample preparation and instrumentation has led to topographs that reveal subnanometer details and the surface dynamics of biomolecules. Tethering single molecules between a support and a retracting AFM tip produces force–extension curves, giving information about the mechanical stability of secondary structural elements. For both imaging and force spectroscopy, the cantilever and its tip are critical:the mechanical properties of the cantilever dictate the force sensitivity and the scanning speed, whereas the tip shape determines the achievable lateral resolution.

Clathrin-Inspired Coating and Stabilization of Liposomes by DNA Self-Assembly

2019 ◽  
Author(s):  
Kevin N. Baumann ◽  
Luca Piantanida ◽  
Javier García-Nafría ◽  
Diana Sobota ◽  
Kislon Voïtchovsky ◽  
...  
Keyword(s):  
Self Assembly ◽  
Mechanical Stability ◽  
Lipid Vesicles ◽  
The Self ◽  
Force Microscopy ◽  
Atomic Force ◽  
Cryo Electron Microscopy ◽  
Further Development ◽  
Liposome Surface ◽  
Dna Coating

The self-assembly of the protein clathrin on biological membranes facilitates essential processes of endocytosis in biological systems and has provided a source of inspiration for materials design by the highly ordered structural appearance. By mimicking the architecture of clathrin self-assemblies to coat liposomes with biomaterials, new classes of hybrid carriers can be derived. Here we present a method for fabricating DNA-coated liposomes by hydrophobically anchoring and subsequently growing a DNA network on the liposome surface which structurally mimics clathrin assemblies. Dynamic light scattering (DLS), ζ-potential and cryo-electron microscopy (cryo-EM) measurements independently demonstrate successful DNA coating. Nanomechanical measurements conducted with atomic force microscopy (AFM) show that the DNA coating enhances the mechanical stability of the liposomes relative to uncoated ones. Furthermore, we provide the possibility to reverse the coating process by triggering the disassembly of the DNA coating through a toehold-mediated displacement reaction. Our results describe a straightforward, versatile, and reversible approach for coating and stabilizing lipid vesicles by an interlaced DNA network. This method has potential for further development towards the ordered arrangement of tailored functionalities on the surfaces of liposomes and for applications as hybrid nanocarrier.

scholarly journals 3D phonon microscopy with sub-micron axial-resolution

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Richard J. Smith ◽  
Fernando Pérez-Cota ◽  
Leonel Marques ◽  
Matt Clark
Keyword(s):  
Signal To Noise Ratio ◽  
Single Cells ◽  
Optical Resolution ◽  
Acquisition Time ◽  
Label Free ◽  
Axial Resolution ◽  
Force Microscopy ◽  
Atomic Force ◽  
Accuracy And Precision ◽  
Good Agreement

AbstractBrillouin light scattering (BLS) is an emerging method for cell imaging and characterisation. It allows elasticity-related contrast, optical resolution and label-free operation. Phonon microscopy detects BLS from laser generated coherent phonon fields to offer an attractive route for imaging since, at GHz frequencies, the phonon wavelength is sub-optical. Using phonon fields to image single cells is challenging as the signal to noise ratio and acquisition time are often poor. However, recent advances in the instrumentation have enabled imaging of fixed and living cells. This work presents the first experimental characterisation of phonon-based axial resolution provided by the response to a sharp edge. The obtained axial resolution is up to 10 times higher than that of the optical system used to take the measurements. Validation of the results are obtained with various polymer objects, which are in good agreement with those obtained using atomic force microscopy. Edge localisation, and hence profilometry, of a phantom boundary is measured with accuracy and precision of approximately 60 nm and 100 nm respectively. Finally, 3D imaging of fixed cells in culture medium is demonstrated.

Approximate Real-Time Force Spectroscopy Within Amplitude-Modulation Atomic Force Microscopy Topographical Imaging Using Few Harmonics and Fourier Methods

2021 ◽  
Author(s):  
Berkin Uluutku ◽  
Santiago D. Solares
Keyword(s):  
Atomic Force Microscopy ◽  
Signal To Noise Ratio ◽  
Gaussian Model ◽  
Force Curve ◽  
Simple Method ◽  
Force Microscopy ◽  
Electromagnetic Interactions ◽  
Atomic Force ◽  
Mechanical Deformations ◽  
Specialized Hardware

Abstract Quantitative measurement of the probe-sample interaction forces as a function of distance and time during imaging has been at the forefront of atomic force microscopy (AFM) research. This type of information is extremely valuable for understanding the material response to a variety of stimuli and interactions, such as mechanical deformations that vary in magnitude and rate of application, chemical interactions, or electromagnetic interactions. A variety of methods for performing such measurements simultaneously with topographical imaging is available, including methods based on Fourier analysis. Within these methods, reconstruction of the tip-sample force curve generally requires measurement of a large number of harmonics of the probe oscillation, which presents challenges such as the need for specialized hardware, low signal-to-noise ratio, and the need for extensive user expertise. In this paper, we present a simple method to perform a Gaussian-model-based fit of the tip-sample force curve across the surface, simultaneously with imaging, which requires measurement of only the first two or three harmonics for elastic materials. While such an approach only offers an approximate representation of the force curve, it can be highly accurate and fast, and has low instrumentation requirements, such that it can be relatively simple to implement on most commercial AFM setups.

scholarly journals Tunable control of antibody immobilization using electric field

2015 ◽  
Vol 112 (7) ◽  
pp. 1995-1999 ◽  
Author(s):  
Sam Emaminejad ◽  
Mehdi Javanmard ◽  
Chaitanya Gupta ◽  
Shuai Chang ◽  
Ronald W. Davis ◽  
...  
Keyword(s):  
Solid State ◽  
Electric Field ◽  
Signal To Noise Ratio ◽  
Physical Adsorption ◽  
Antibody Immobilization ◽  
Proof Of Concept ◽  
Force Microscopy ◽  
Atomic Force ◽  
Sensing Platforms ◽  
Glass Surfaces

The controlled immobilization of proteins on solid-state surfaces can play an important role in enhancing the sensitivity of both affinity-based biosensors and probe-free sensing platforms. Typical methods of controlling the orientation of probe proteins on a sensor surface involve surface chemistry-based techniques. Here, we present a method of tunably controlling the immobilization of proteins on a solid-state surface using electric field. We study the ability to orient molecules by immobilizing IgG molecules in microchannels while applying lateral fields. We use atomic force microscopy to both qualitatively and quantitatively study the orientation of antibodies on glass surfaces. We apply this ability for controlled orientation to enhance the performance of affinity-based assays. As a proof of concept, we use fluorescence detection to indirectly verify the modulation of the orientation of proteins bound to the surface. We studied the interaction of fluorescently tagged anti-IgG with surface immobilized IgG controlled by electric field. Our study demonstrates that the use of electric field can result in more than 100% enhancement in signal-to-noise ratio compared with normal physical adsorption.

Diabetes increases stiffness of live cardiomyocytes measured by atomic force microscopy nanoindentation

2014 ◽  
Vol 307 (10) ◽  
pp. C910-C919 ◽  
Author(s):  
Juan C. Benech ◽  
Nicolás Benech ◽  
Ana I. Zambrana ◽  
Inés Rauschert ◽  
Verónica Bervejillo ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Physiological Condition ◽  
Diabetic Mice ◽  
Cell Nuclei ◽  
Buffer Solution ◽  
Isolated Cardiomyocytes ◽  
Force Microscopy ◽  
Atomic Force ◽  
Myocardial Fibers

Stiffness of live cardiomyocytes isolated from control and diabetic mice was measured using the atomic force microscopy nanoindentation method. Type 1 diabetes was induced in mice by streptozotocin administration. Histological images of myocardium from mice that were diabetic for 3 mo showed disorderly lineup of myocardial cells, irregularly sized cell nuclei, and fragmented and disordered myocardial fibers with interstitial collagen accumulation. Phalloidin-stained cardiomyocytes isolated from diabetic mice showed altered (i.e., more irregular and diffuse) actin filament organization compared with cardiomyocytes from control mice. Sarco/endoplasmic reticulum Ca2+-ATPase (SERCA2a) pump expression was reduced in homogenates obtained from the left ventricle of diabetic animals compared with age-matched controls. The apparent elastic modulus (AEM) for live control or diabetic isolated cardiomyocytes was measured using the atomic force microscopy nanoindentation method in Tyrode buffer solution containing 1.8 mM Ca2+ and 5.4 mM KCl (physiological condition), 100 nM Ca2+ and 5.4 mM KCl (low extracellular Ca2+ condition), or 1.8 mM Ca2+ and 140 mM KCl (contraction condition). In the physiological condition, the mean AEM was 112% higher for live diabetic than control isolated cardiomyocytes (91 ± 14 vs. 43 ± 7 kPa). The AEM was also significantly higher in diabetic than control cardiomyocytes in the low extracellular Ca2+ and contraction conditions. These findings suggest that the material properties of live cardiomyocytes were affected by diabetes, resulting in stiffer cells, which very likely contribute to high diastolic LV stiffness, which has been observed in vivo in some diabetes mellitus patients.

scholarly journals UV Irradiated Graphene-Based Nanocomposites: Change in the Mechanical Properties by Local HarmoniX Atomic Force Microscopy Detection

Materials ◽  
2019 ◽  
Vol 12 (6) ◽  
pp. 962 ◽  
Author(s):  
Liberata Guadagno ◽  
Carlo Naddeo ◽  
Marialuigia Raimondo ◽  
Vito Speranza ◽  
Roberto Pantani ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
Atomic Force Microscopy ◽  
Mechanical Stability ◽  
Uv Light ◽  
Ultra Violet ◽  
Graphene Nanoplatelets ◽  
Free Standing ◽  
Force Microscopy ◽  
Atomic Force ◽  
Acquisition Mode

Epoxy based coatings are susceptible to ultra violet (UV) damage and their durability can be significantly reduced in outdoor environments. This paper highlights a relevant property of graphene-based nanoparticles: Graphene Nanoplatelets (GNPs) incorporated in an epoxy-based free-standing film determine a strong decrease of the mechanical damages caused by UV irradiation. The effects of UV light on the morphology and mechanical properties of the solidified nanocharged epoxy films are investigated by Atomic Force Microscopy (AFM), in the acquisition mode “HarmoniX.” Nanometric-resolved maps of the mechanical properties of the multi-phase material evidence that the incorporation of low percentages, between 0.1% and 1.0% by weight, of graphene nanoplatelets (GNPs) in the polymeric film causes a relevant enhancement in the mechanical stability of the irradiated films. The beneficial effect progressively increases with increasing GNP percentage. The paper also highlights the potentiality of AFM microscopy, in the acquisition mode “HarmoniX” for studying multiphase polymeric systems.

scholarly journals Functional Mapping of Single Molecules and Gels using Atomic Force Microscopy

2009 ◽  
Vol 96 (3) ◽  
pp. 644a
Author(s):  
Sarah Baos ◽  
Monica Berry ◽  
Peter Heard ◽  
Debra Brayshaw ◽  
Terence J. McMaster
Keyword(s):  
Atomic Force Microscopy ◽  
Single Molecules ◽  
Functional Mapping ◽  
Force Microscopy ◽  
Atomic Force

Cytoskeletal Dynamics of Neurons Measured by Combined Fluorescence and Atomic Force Microscopy

MRS Advances ◽  
2018 ◽  
Vol 3 (26) ◽  
pp. 1463-1468 ◽  
Author(s):  
Peter Moore ◽  
Cristian Staii
Keyword(s):  
Mechanical Properties ◽  
Atomic Force Microscopy ◽  
Ambient Temperature ◽  
Cortical Neurons ◽  
Mechanical Stability ◽  
Neuronal Cells ◽  
Force Microscopy ◽  
Atomic Force ◽  
Key Factor ◽  
External Forces

ABSTRACTMechanical properties of neurons represent a key factor that determines the functionality of neuronal cells and the formation of neural networks. The main source of mechanical stability for the cell is a biopolymer network of microtubules and actin filaments that form the main components of the cellular cytoskeleton. This biopolymer network is responsible for the growth of neuronal cells as they extend neurites to connect with other neurons, forming the nervous system. Here we present experimental results that combine atomic force microscopy (AFM) and fluorescence microscopy to produce systematic, high-resolution elasticity and fluorescence maps of cortical neurons. This approach allows us to apply external forces to neurons, and to monitor the dynamics of the cell cytoskeleton. We measure how the elastic modulus of neurons changes upon changing the ambient temperature, and identify the cytoskeletal components responsible for these changes. These results demonstrate the importance of taking into account the effect of ambient temperature when measuring the mechanical properties of cells.

Imaging of reconstituted biological channels at molecular resolution by atomic force microscopy

1993 ◽  
Vol 265 (3) ◽  
pp. C851-C856 ◽  
Author(s):  
R. Lal ◽  
H. Kim ◽  
R. M. Garavito ◽  
M. F. Arnsdorf
Keyword(s):  
Atomic Force Microscopy ◽  
High Resolution ◽  
Internal Control ◽  
Imaging Method ◽  
Buffer Solution ◽  
Electron Microscopic ◽  
Ompf Porin ◽  
Force Microscopy ◽  
Atomic Force ◽  
Rectangular Pattern

Using atomic force microscopy (AFM), we obtained high-resolution surface images of the bacterial outer membrane channels Escherichia coli OmpF porin and Bordetella pertussis porin that were reconstituted in artificial bilayer membranes as two-dimensional crystalline arrays. These porins were chosen because they are among the most extensively studied proteins of this type and are known for their well-defined crystalline nature in the native membrane. Such reconstituted membrane proteins are ideal specimens to assess the suitability and resolution of AFM for imaging biomembranes and associated proteins. Although OmpF porin often showed a mixed pattern of rectangular and hexagonal arrays with approximately 8.4 x 9.8- and approximately 7.2-nm-spacings, respectively, B. pertussis porin showed mostly a rectangular pattern with an approximately 7.9 x 13.8-nm spacing. The packing patterns of the E. coli OmpF porin in the membrane are very close to those found in electron-microscopic studies. When B. pertussis porin was imaged in a buffer solution, its trimeric subunits were apparently resolved, and the surface of each monomer revealed beadlike structures. This is the first report of such a high-resolution structural analysis of B. pertussis porin by any imaging method. We also imaged the lipid bilayer itself as an internal control for imaging and to further ascertain the resolution. Individual polar head groups of bilayer lipid molecules were resolved, suggesting the intrinsic resolution of AFM for bioimaging.

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