Atomic Force Microscopy Images Research Articles

In situ creation of aramid fiber surface coating with polymerizable deep eutectic solvents and their interfacial bonding properties with rubber

AbstractThe coating was created on the surface of aramid fibers (AFs) by solution dip‐coating and in situ UV curable method, using a polymerizable deep eutectic solvent (PDES), dissolved poly(ethylene imine), and poly(ethylene glycol) diacrylate (PEGDA). Fourier transform infrared spectroscopy and thermogravimetry confirmed the successful construction of the coating on the AF surface, which accounted for 12%–17% of the total fiber mass. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) images revealed a 2 μm increase in fiber diameter and a maximum fiber surface average roughness (Ra) of 263.7 nm when the polyethyleneimine content was 0.5% (AF‐PDES‐0.5). Based on these results, H pull‐out samples of fiber/rubber composites were fabricated. The results indicated that the H pull‐out force of AF‐PDES‐0.5 samples increased by 109.74% than that of untreated fiber, and reached 12.92 N of the maximum strength. In summary, the method is simple, rapid, non‐polluting, capable of continuous, and intelligent construction of coatings on the surface of AF. Thus, making it highly promising for application in the field of aviation tires.Highlights Using PDES, green, and low‐carbon preparation processes. Solution dip‐coating and UV reaction for improved reaction speed and workability. Quantitative of interface thickness and modulus using EDS and AFM.

Effect of Substrate Temperature and Deposition Power on the Surface Morphology and Optical Properties of ZnO:Mg Thin Films Deposited by DC Magnetron Sputtering

Magnesium doped zinc oxide (ZnO:Mg) thin films were deposited on soda lime glass slides by DC magnetron sputtering method. Atomic force microscope (AFM), and UV/VIS spectrophotometer were used to investigate the effect of sputtering power and substrate temperature on the surface morphology and optical properties of ZnO:Mg thin films. AFM images revealed that sputtering power and deposition temperature have significant influence on surface morphology of the ZnO:Mg thin films. For all sputtering powers and substrate temperatures investigated, ZnO:Mg films had peak transmittance above 85%. Samples deposited at 110 W sputtering power and 450 °C substrate temperature showed the best peak transmittance of > 90% at 560 nm (visible range). Optical band gap of ZnO:Mg films was in the range of 3.44– 3.69 eV depending on the substrate temperature. The results indicated the potential of the films for transparent conductor applications.

LLDPE film development surface with nucleating agents to reduce fat stains from food adherence on the film surface for easy rinsing before recycling

The purpose of this research was to enhance the surface of linear low-density polyethylene (LLDPE) film by using single and hybrid nucleating agents to reduce fat stains from food adherence on the film surface for easy rinsing before recycling, consequently reducing energy consumption before recycling. The process begins with the preparation of a high-concentration masterbatch of LLDPE with nucleating agents and polyethylene glycol (PEG), followed by blown film and test analysis. The results showed that both single and hybrid nucleating agents resulted in substantial modifications in the surface characteristics of the film. The altered surface texture of the film makes fat stains easy to remove. In addition, the use of hybrid nucleating agents resulted in considerable roughness on the film's surface. It can calculate the average roughness (Ra) from atomic force microscopy (AFM) images, which is very low at 4.7 nm, and the roughness is consistent. The contact angle was 99.17 degrees, and the percentage of crystallinity was clearly higher than that of a single nucleating agent. As a result, the fat stains on the film surface are easier to remove. The visual appearance, morphology, functional group, and mechanical properties of the films, on the other hand, were not significantly different. In summary, using a nucleating agent helped to rinse fat stains from the film easily.

Quantitative analysis of [formula omitted] thin film micrographs with machine learning

Isolating the features associated with different materials growth conditions is important to facilitate the tuning of these conditions for effective materials growth and characterization. This study presents machine learning models for classifying atomic force microscopy (AFM) images of thin film MoS2 based on their growth temperatures. By employing nine different algorithms and leveraging transfer learning through a pretrained ResNet model, we identify an effective approach for accurately discerning the characteristics related to growth temperature within the AFM micrographs. Robust models with test accuracies of up to 70% were obtained, with the best performing algorithm being an end-to-end ResNet fine-tuned on our image domain. Class activation maps and occlusion attribution reveal that crystal quality and domain boundaries play crucial roles in classification, with models exhibiting the ability to identify latent features that humans could potentially miss. Overall, the models demonstrated high accuracy in identifying thin films grown at different temperatures despite limited and imbalanced training data as well as variation in growth parameters besides temperature, showing that our models and training protocols are suitable for this and similar predictive tasks for accelerated 2D materials characterization.

Self-Healing Composite Coating Fabricated with a Cystamine Cross-Linked Cellulose Nanocrystal-Stabilized Pickering Emulsion.

A gelled Pickering emulsion system was fabricated by first stabilizing linseed oil droplets in water with dialdehyde cellulose nanocrystals (DACNCs) and then cross-linking with cystamine. Cross-linking of the DACNCs was shown to occur by a reaction between the amine groups on cystamine and the aldehyde groups on the CNCs, causing gelation of the nanocellulose suspension. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy were used to characterize the cystamine-cross-linked CNCs (cysCNCs), demonstrating their presence. Transmission electron microscopy images evidenced that cross-linking between cysCNCs took place. This cross-linking was utilized in a linseed oil-in-water Pickering emulsion system, creating a novel gelled Pickering emulsion system. The rheological properties of both DACNC suspensions and nanocellulose-stabilized Pickering emulsions were monitored during the cross-linking reaction. Dynamic light scattering and confocal laser scanning microscopy (CLSM) of the Pickering emulsion before gelling imaged CNC-stabilized oil droplets along with isolated CNC rods and CNC clusters, which had not been adsorbed to the oil droplet surfaces. Atomic force microscopy imaging of the air-dried gelled Pickering emulsion also demonstrated the presence of free CNCs alongside the oil droplets and the cross-linked CNC network directly at the oil-water interface on the oil droplet surfaces. Finally, these gelled Pickering emulsions were mixed with poly(vinyl alcohol) solutions and fabricated into self-healing composite coating systems. These self-healing composite coatings were then scratched and viewed under both an optical microscope and a scanning electron microscope before and after self-healing. The linseed oil was demonstrated to leak into the scratches, healing the gap automatically and giving a practical approach for a variety of potential applications.

Optical, structural, morphological, and gas sensing properties of Mg-doped α-Fe2O3 thin films deposited by RF and DC magnetron Co-sputtering technique

In this research, Mg-doped hematite (α-Fe2O3) thin films with different Mg-doping concentrations were synthesized on glass substrates using the direct current (DC) and radio frequency (RF) magnetron co-sputtering technique, and the changes of some physical properties due to the concentration of the dopant were investigated. The optical, structural, morphological, and elemental properties of the obtained Mg-doped α-Fe2O3 thin films were determined by X-ray diffraction (XRD) analysis, UV–Vis spectroscopy, Raman spectroscopy, scanning electron microscopy (SEM), dispersive X-ray spectroscopy and atomic force microscopy (AFM). XRD analysis revealed that the investigated thin films have a rhombohedral crystal structure. The deposition of films at different DC sputtering voltages caused significant variations in stoichiometry and nanostructure. The thin films' band gap energy values were estimated based on absorption measurements, and results ranged from 2.15 eV to 2.69 eV. Raman peaks were observed between 218 cm−1 and 1305 cm−1. The thin films exhibit uniform surface morphology throughout the substrate according to the SEM images. The low RMS roughness values obtained from AFM images showed that the surfaces of Mg:Fe2O3 thin films are smooth. The Mg-doped α-Fe2O3 thin film doped by 150 W DC voltage exhibited a good gas-sensing response at 300 °C. A remarkably quick response/recovery time was achieved.

Molecular identification with atomic force microscopy and conditional generative adversarial networks

Frequency modulation (FM) atomic force microscopy (AFM) with metal tips functionalized with a CO molecule at the tip apex (referred as High-Resolution AFM, HR-AFM) has provided access to the internal structure of molecules with totally unprecedented resolution. We propose a model to extract the chemical information from those AFM images in order to achieve a complete identification of the imaged molecule. Our Conditional Generative Adversarial Network (CGAN) converts a stack of constant-height HR-AFM images at various tip-sample distances into a ball-and-stick depiction, where balls of different color and size represent the chemical species and sticks represent the bonds, providing complete information on the structure and chemical composition. The CGAN has been trained and tested with the QUAM-AFM data set, that contains simulated AFM images for a collection of 686000 organic molecules that include all the chemical species relevant in organic chemistry. Tests with a large set of theoretical images and few experimental examples demonstrate the accuracy and potential of our approach for molecular identification.

Mycogenic synthesis of iron nanoparticles using thermophilic mould Myceliophthora thermophila and their applicability in environmental remediation

Culture extract of thermophilic mould Myceliophthora thermophila BJTLRMDU7 played important role in the green synthesis of iron nanoparticles (FeNPs) using ferrous sulphate (FeSO4). The mould was grown in potato dextrose broth (PDB) at 45 °C and 200 rpm for 72 h. A colour change from transparent to reddish brown showed the synthesis of FeNPs due to the surface plasmon resonance by the fungal culture extract. The process for the synthesis of nanoparticles was optimized using different reaction conditions. Biogenic iron nanoparticles were synthesized maximally at 50 °C and pH 5.0 after 24 h using 2 mM salt solution and one ml culture extract. Furthermore, formation of iron nanoparticles was significantly accelerated in the presence of light. The absorption peak was maximum around 250–350 nm in UV–vis spectrum. The absorption peaks of Fourier-transform infrared spectroscopy (FTIR) at 619.99 cm−1, 793.57 cm−1, 882.32 cm−1, 1121.46 cm−1, 1627.71 cm−1, 2923.17 cm−1 and 3430.70 cm−1, indicated functional groups of active biomolecules involved in the synthesis of FeNPs. Atomic force microscopy (AFM) images of green synthesized iron nanoparticles indicated the average size of 80 nm, while X-ray diffraction (XRD) data revealed the crystalline nature of FeNPs. These FeNPs efficiently reduced p-nitrophenol (PNP) into p-aminophenol (PAP), which was further confirmed by thin layer chromatography. Biogenic FeNPs showed 96 and 35 % decolourization of bromphenol blue and malachite green, respectively. Further, addition of H2O2 enhanced the decolourization of malachite green up to 96 %. Biogenic FeNPs synthesized from culture extract of a thermophilic mould have been found effective in remediation of environmental pollutants released from various industries.

Evaluation of antimicrobial photodynamic action of a pluronic and pectin based film loaded with methylene blue against methicillin resistant Staphylococcus aureus

Antimicrobial wound dressings play a crucial role in treatment of wound infections. However, existing commercial options fall short due to antibiotic resistance and the limited spectrum of activity of newly emerging antimicrobials against bacteria that are frequently encountered in wound infections. Antimicrobial photodynamic therapy (aPDT) is very promising alternative therapeutic approach against antibiotic resistant microbes such as methicillin resistant Staphylococcus aureus (MRSA). However, delivery of the photosensitizer (PS) homogeneously to the wound site is a challenge. Though polymeric wound dressings based on synthetic and biopolymers are being explored for aPDT, there is paucity of data regarding their in vivo efficacy. Moreover, there are no studies on use of PS loaded, pluoronic (PL) and pectin (PC) based films for aPDT. We report development of a polymeric film for potential use in aPDT. The film was prepared using PL and PC via solvent casting approach and impregnated with methylene blue (MB) for photodynamic inactivation of MRSA in vitro and in vivo. Atomic force microscopic imaging of the films yielded vivid pictures of surface topography, with rough surfaces, pores, and furrows. The PL:PC ratio (2:3) was optimized that would result in an intact film but exhibit rapid release of MB in time scale suitable for aPDT. The film showed good antibacterial activity against planktonic suspension, biofilm of MRSA upon exposure to red light. Investigations on MRSA infected excisional wounds of mice reveal that topical application of MB loaded film for 30 min followed by red light exposure for 5 min (fluence; ∼30 J cm−2) or 10 min (fluence; ∼60 J cm−2) reduces ∼80% or ∼92% of bioburden, respectively. Importantly, the film elicits no significant cytotoxicity against keratinocytes and human adipose derived mesenchymal stem cells. Taken together, our data demonstrate that PS-loaded PL-PC based films are a promising new tool for treatment of MRSA infected wounds.

Dewetting upside-down: two-sided solid state dewetting of thin gold film on soft KBr substrate

Abstract We deposited a 30 nm-thick Au film on single crystalline KBr substrate and studied the solid state dewetting behavior of the film at a temperature of 350 °C. At this temperature, the ions of the KBr compound exhibit significant mobility along the Au–KBr interface, which affects the morphology and kinetics of the solid state dewetting. We performed statistical morphology analysis of the Au–KBr interface by selectively dissolving the KBr substrate after the dewetting heat treatments and subsequent atomic force microscopy imaging of the “upside-down” oriented Au film. We demonstrated that atomic mobility at the interface leads to embedding of the partially dewetted Au film into the KBr substrate. We proposed a quantitative model of the shape evolution of a disc-shaped Au particle on the KBr substrate under the condition of finite interface mobility of the substrate species. The model predictions were consistent with the experimentally observed sinking rates of Au nanostructures.

Atom Filtering Algorithm and GPU-Accelerated Calculation of Simulation Atomic Force Microscopy Images

Simulation of atomic force microscopy (AFM) computationally emulates experimental scanning of a biomolecular structure to produce topographic images that can be correlated with measured images. Its application to the enormous amount of available high-resolution structures, as well as to molecular dynamics modelling data, facilitates the quantitative interpretation of experimental observations by inferring atomistic information from resolution-limited measured topographies. The computation required to generate a simulated AFM image generally includes the calculation of contacts between the scanning tip and all atoms from the biomolecular structure. However, since only contacts with surface atoms are relevant, a filtering method shall highly improve the efficiency of simulated AFM computations. In this report, we address this issue and present an elegant solution based on graphics processing unit (GPU) computations that significantly accelerates the computation of simulated AFM images. This method not only allows for the visualization of biomolecular structures combined with ultra-fast synchronized calculation and graphical representation of corresponding simulated AFM images (live simulation AFM), but, as we demonstrate, it can also reduce the computational effort during the automatized fitting of atomistic structures into measured AFM topographies by orders of magnitude. Hence, the developed method will play an important role in post-experimental computational analysis involving simulated AFM, including expected applications in machine learning approaches. The implementation is realized in our BioAFMviewer software (ver. 3) package for simulated AFM of biomolecular structures and dynamics.

Sputter grown CuO thin films: Impact of growth pressure and annealing temperature on their microstructural architectures

High-quality copper oxide (CuO) thin films were deposited on the silicon (Si) substrate at the room temperature using the physical vapour deposition (PVD) technique named radio frequency (RF) sputtering. The copper-oxide thin-films were single crystalline and of uniform thickness. Subsequently, the influence of growth pressure (low gas pressure - 3 mTorr and high gas pressure - 100 mTorr) and post growth annealing at different temperatures (300 °C to 700 °C) were investigated to understand the microstructural and morphological changes of the thin film. With the influence of growth pressure and post thermal annealing temperature, significant changes in crystallinity, surface roughness, and surface oxidation rate of the CuO thin film were detected, which were adequately analyzed via several characterization techniques. X-ray diffraction (XRD) patterns revealed the phase formation with good crystallinity of the film, which is substantiated by Raman spectroscopy and X-ray photoelectron spectroscopy (XPS) characterization. Atomic force microscopy (AFM) images disclosed that the surface roughness of the film and grain size. By gaining insights into the structural and surface properties of CuO/Si thin films, this research presents new prospects for tuning of CuO phases, structures, and compositions for multifunctional applications.

Combined Role of Interleukin-15 Stimulated Natural Killer Cell-Derived Extracellular Vesicles and Carboplatin in Osimertinib-Resistant H1975 Lung Cancer Cells with EGFR Mutations.

In this study, we evaluated IL-15 stimulated natural killer cell-derived EVs (NK-EVs) as therapeutic agents in vitro and in vivo in Osimertinib-resistant lung cancer (H1975R) with EGFR mutations (L858R) in combination with carboplatin (CBP). NK-EVs were isolated by ultracentrifugation and characterized by nanoparticle tracking analysis, and atomic force microscopy imaging revealed vesicles with a spherical form and sizes meeting the criteria of exosomal EVs. Further, Western blot studies demonstrated the presence of regular EV markers along with specific NK markers (perforin and granzyme). EVs were also characterized by proteomic analysis, which demonstrated that EVs had proteins for natural killer cell-mediated cytotoxicity (Granzyme B) and T cell activation (perforin and plastin-2). Gene oncology analysis showed that these differentially expressed proteins are involved in programmed cell death and positive regulation of cell death. Further, isolated NK-EVs were cytotoxic to H1975R cells in vitro in 2D and 3D cell cultures. CBP's IC50 was reduced by approximately in 2D and 3D cell cultures when combined with NK-EVs. The EVs were then combined with CBP and administered by i.p. route to H1975R tumor xenografts, and a significant reduction in tumor volume in vivo was observed. Our findings show for the first time that NK-EVs target the PD-L1/PD-1 immunological checkpoint to induce apoptosis and anti-inflammatory response by downregulation of SOD2, PARP, BCL2, SET, NF-κB, and TGF-ß. The ability to isolate functional NK-EVs on a large scale and use them with platinum-based drugs may lead to new clinical applications. The results of the present study suggest the possibility of the combination of NK-cell-derived EVs and CBP as a viable immunochemotherapeutic strategy for resistant cancers.

Thermally evaporated vanadium-based phthalocyanine for low moisture detection in humidity sensors

Aluminium / vanadyl 3,10,17,24-tetra-tert-butyl-1,8,15,22-tetrakis(dimethylamino)− 29 H,31 H-phthalocyanine (VTP) / Aluminium, (Al/VTP/Al) has been successfully studied for capacitive humidity sensing. The metal-containing vanadium central atom in the metal phthalocyanine has contributed towards excellent capacitance response at low humidity conditions as low as 10%RH. In addition, high surface roughness revealed by atomic force microscopy (AFM) images has also given insight into the low hysteresis lag. The film's large surface area helped prevent the water molecules from accumulating in the VTP pores and capillaries. Grazing-incidence wide and small-angle X-ray scattering (GIWAXS-GISAXS) indicates some face-on crystallite orientation produced by the partially ordered film, impacting the transient performance of the sensor.

Probing the interactions of the HIV-1 matrix protein-derived polybasic region with lipid bilayers: insights from AFM imaging and force spectroscopy.

The human immunodeficiency virus type 1 (HIV-1) matrix protein contains a highly basic region, MA-HBR, crucial for various stages of viral replication. To elucidate the interactions between the polybasic peptide MA-HBR and lipid bilayers, we employed liquid-based atomic force microscopy (AFM) imaging and force spectroscopy on lipid bilayers of differing compositions. In 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayers, AFM imaging revealed the formation of annulus-shaped protrusions upon exposure to the polybasic peptide, accompanied by distinctive mechanical responses characterized by enhanced bilayer puncture forces. Importantly, our AFM-based force spectroscopy measurements unveiled that MA-HBR induces interleaflet decoupling within the cohesive bilayer organization. This is evidenced by a force discontinuity observed within the bilayer's elastic deformation regime. In POPC/cholesterol bilayers, MA-HBR caused similar yet smaller annular protrusions, demonstrating an intriguing interplay with cholesterol-rich membranes. In contrast, in bilayers containing anionic 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine (POPS) lipids, MA-HBR induced unique annular protrusions, granular nanoparticles, and nanotubules, showcasing its distinctive effects in anionic lipid-enriched environments. Notably, our force spectroscopy data revealed that anionic POPS lipids weakened interleaflet adhesion within the bilayer, resulting in interleaflet decoupling, which potentially contributes to the specific bilayer perturbations induced by MA-HBR. Collectively, our findings highlight the remarkable variations in how the polybasic peptide, MA-HBR, interacts with lipid bilayers of differing compositions, shedding light on its role in host membrane restructuring during HIV-1 infection.

Investigation on the electrocatalytic oxidation of alcohol using zinc oxide thin films deposited by pulsed electrodeposition on an indium tin oxide surface

To perform electro-catalytic oxidation of alcohols in an alkaline media, zinc oxide (ZnO) thin films were effectively deposited on indium tin oxide (ITO) substrates using the pulsed electrodeposition (PE) method in a zinc nitrate aqueous solution. Analyses of microstructural, photoelectrochemical, and optical characteristics of ZnO thin films were performed as a function of pulsed electrodeposition parameters. XRD analysis was applied to determine structural properties, and SEM and AFM analysis were used to investigate morphological characteristics. X-ray diffraction analyses revealed that the produced films were polycrystalline and had a (002) preferentially oriented, hexagonal wurtzite structure. The morphology of ZnO has improved in the direction of nanorod films, as evidenced by scanning electron microscopy (SEM) and atomic force microscopy (AFM) images. UV transmittance data was used using Tauc's relationship to determine the film's bandgap to be 3.26 eV. The photocurrent response shows that ZnO films have high values, at 305.17 A/cm2, and good optical characteristics. The electrocatalytic oxidation of methanol was finally evaluated on the films. After being optimized in the pulsed electrodeposition mode, ZnO thin film characteristics and methanol electrocatalytic oxidation both saw substantial improvements.

N-Formylation modifies membrane damage associated with PSMα3 interfacial fibrillation.

The virulence of Staphylococcus aureus, a multi-drug resistant pathogen, notably depends on the expression of the phenol soluble modulins α3 (PSMα3) peptides, able to self-assemble into amyloid-like cross-α fibrils. Despite remarkable advances evidencing the crucial, yet insufficient, role of fibrils in PSMα3 cytotoxic activities towards host cells, the relationship between its molecular structures, assembly propensities, and modes of action remains an open intriguing problem. In this study, combining atomic force microscopy (AFM) imaging and infrared spectroscopy, we first demonstrated in vitro that the charge provided by the N-terminal capping of PSMα3 alters its interactions with model membranes of controlled lipid composition without compromising its fibrillation kinetics or morphology. N-formylation eventually dictates PSMα3-membrane binding via electrostatic interactions with the lipid head groups. Furthermore, PSMα3 insertion within the lipid bilayer is favoured by hydrophobic interactions with the lipid acyl chains only in the fluid phase of membranes and not in the gel-like ordered domains. Strikingly, our real-time AFM imaging emphasizes how intermediate protofibrillar entities, formed along PSMα3 self-assembly and promoted at the membrane interface, likely disrupt membrane integrity via peptide accumulation and subsequent membrane thinning in a peptide concentration and lipid-dependent manner. Overall, our multiscale and multimodal approach sheds new light on the key roles of N-formylation and intermediate self-assembling entities, rather than mature fibrils, in dictating deleterious interactions of PSMα3 with membrane lipids, likely underscoring its ultimate cellular toxicity in vivo, and in turn S. aureus pathogenesis.

Imaging of Mucin Networks with Atomic Force Microscopy.

Mucin networks serve as the structural scaffold of mucus and play a significant role in determining its biophysical properties. Thus, characterizing the organization, macromolecular structure, and interactions within these networks is a key step in understanding the parameters that govern mucus functionality in both health and disease. Atomic force microscopy (AFM) is uniquely suited to study mucin networks; AFM can clearly resolve nanometer-sized features, does not require fixation or metallization, and can be performed in air or aqueous solutions. In this chapter we describe protocols to image mucin networks using AFM. First, we describe two protocols to enrich and isolate mucin samples in preparation for AFM imaging. Next, we detail a protocol to deposit the samples onto a mica substrate. Finally, we give general tips to optimize and troubleshoot AFM imaging of mucin networks.

Quantifying ABCA1/apoA-1 Signaling Pathways with AFM Imaging and Lipidomic Analysis.

The role of lipid metabolic pathways in the pathophysiology of primary open-angle glaucoma (POAG) has been thoroughly elucidated, with pathways involved in lipid-related disorders such as hypercholesterolemia and hyperlipoprotein accumulation being of particular interest. The ABCA1/apoA-1 transduction pathway moderates reverse cholesterol transport (RCT), facilitating the transport of free cholesterol (FC) and phospholipids (PL) and preventing intracellular lipid aggregates in retinal ganglion cells (RGCs) due to excess FCs and PLs. A deficiency of ABCA1 transporters, and thus, dysregulation of the ABCA1/apoA-1 transduction pathway, may potentiate cellular lipid accumulation, which affects the structural and mechanical features of the cholesterol-rich RGC membranes. Atomic force microscopy (AFM) is a cutting-edge imaging technique suitable for imaging topographical surfaces of a biological specimen and determining its mechanical properties and structural features. The versatility and precision of this technique may prove beneficial in understanding the effects of ABCA1/apoA-1 pathway downregulation and decreased cholesterol efflux in RGCs and their membranes. In this protocol, ABCA1-/- RGC mouse models are prepared over the course of 3days and are then compared with non-knockout ABCA1 RGC mouse models through AFM imaging of topographical surfaces to examine the difference in membrane dynamics of knockout vs. non-knockout models. Intracellular and extracellular levels of lipids are quantified through high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS).

Investigating ultra-thin Ag and Au layers using spectrophotometry and AFM imaging

A spectrophotometric method is demonstrated for refractive index and thickness determination of thin and ultrathin metallic films. The method involves a three-layer stack where the metallic layer of interest is deposited on an opaque Si wafer coated with SiO2. This stack creates oscillations in the reflectance spectrum, which are highly sensitive to the index of the metallic film, allowing precise determination of the index of layers down to 1 nm. Experimental index values are given for Ag and Au over the wavelength range of 370–835 nm. These results are correlated with Atomic force microscopy (AFM) images of the films, which reveal dramatic changes in structure for layers of different thickness.