Investigation of human hair fibers using lateral force microscopy

Atomic force microscopy (AFM) and lateral force microscopy (LFM) were used to investigate the morphologic and surface changes associated with various surface modifications to human hair. These included extraction with a series of solvents, bleaching, and treatment with a cationic copolymer. The study assessed the ability of these techniques to distinguish the changes in surface properties, including morphology and friction coefficient, as manifested in changes brought about by the indicated surface modifications. While topographic morphology can easily be investigated with contact AFM. LFM offers an additional tool for probing the surface distribution of oils and waxes. The removal of surface lipids from the fiber surface was accomplished using soxhlet extraction with t-butanol and n-hexane, while the free internal lipids (within the fiber structure) were removed by extraction with a mixture of chloroform and methanol (70:30, v/v). In addition, the surface of hair was modified with the cationic polymer, co(vinyl pyrrolidone-methacrylamidopropyl trimethylammonium chloride [PVP/MAPTAC]), and its distribution on the surface was monitored. Ambient AFM and LFM studies of surface modified and native fibers clearly indicate that when investigated as a function of tip loading force, the different modifications result in changes of the friction coefficient, which increase in this order: native, bleached, solvent extracted, and polymer-treated hair. Friction images show surface variations that are interpreted as areas of varying lipid film coverage. In addition, topographic images of the fibers show the presence of small pores, which become increasingly prevalent upon solvent extraction.     

Atomic force microscopy as a tool for study of human hair

Atomic force microscopy (AFM) is a newly developed microscopic technique that offers high-resolution power, less intrusive measurement, and requires little sample pretreatment for elucidating structures of biological materials in three dimensions and in their natural environment. In this study, AFM has been used not only as an imaging technique for examining human hair structure at high resolution, but also as a tool for quantitative assessment of the effect of treatment in 10 mM phosphate buffered saline of pHs 3.0, 7.0, and 11.0 and heating on human hair structure. It is observed that the hair cuticle is a sensitive indicator of external influences on hair structure, and that its height can be used as a parameter for quantitative assessment. The experimental results obtained show that the swelling of hair caused by the incubation in the buffer decreases with the increase of the pH values and that, depending on the duration of heating, the hair undergoes structural expansion and shrinkage. This study demonstrates that AFM can be used as a valuable alternative to conventional microscopic techniques for hair research.     

Characterization of ultrastructure and collagen composition of the teratoma membrane: Comparison to the amniotic membrane

The structural and morphological properties of the teratoma membrane were investigated to better understand the pathogenesis of ovarian teratomas. A mature cystic teratoma and amnion were obtained from patients who underwent laparoscopic cystectomy and uncomplicated delivery, respectively. The teratoma membrane was divided into three layers according to the results of the histological analysis. Each layer showed distinct morphological properties, including an outer layer that was uniformly arranged, a middle layer with an irregular pattern of fibers, and an inner layer that was structurally dense with a wavy pattern of fibers. The morphology of the layers of the amniotic membrane was the reverse that of the teratoma membrane. In the teratoma membrane, the outer layer was primarily composed of type III collagen and the inner layer had a large amount of type III and IV collagen. The amniotic membrane showed a small amount of type III collagen in the outer layer, whereas the inner layer had large amounts of type I, III, and IV collagen. In the teratoma membrane, the collagen fibrils were arranged regularly in the outer layer, but irregularly in the inner layer. In the amniotic membrane, the arrangement of collagen fibrils was the reverse that of the teratoma membrane. Additionally, the collagen fibrils in the teratoma membrane were thinner than those of the amniotic membrane and had slightly shorter d‐spacing. Two membranes showed the differences in collagen fibril arrangement, which may caused by the different functional roles. Microsc. Res. Tech. 76:432–441, 2013. © 2013 Wiley Periodicals, Inc.     

A quantitative method for analysing AFM images of the outer surfaces of human hair

Cuticle step height is an important parameter for the quantitative assessment of human hair. This paper describes a novel, computational method for the rapid calculation of step heights from atomic force microscope images obtained from large numbers of specimens. Such an approach is necessary to allow a statistical analysis of the inherently wide distribution of cuticle step heights characteristic of a single hair sample. The method described will be of use to cosmetic formulation chemists and forensic scientists and also to dermatologists in the field of disease diagnosis.     

Electron beam lithography-assisted fabrication of Au nano-dot array as a substrate of a correlated AFM and confocal Raman spectroscopy

This paper documents a study of an Au nano-dot array that was fabricated by electron beam lithography on a glass wafer. The patterns that had features of 100nm dots in diameter with a 2-mum pitch comprised a total area of 200x200mum(2). The dot-shaped Cr underlayer was open to the air after developing Poly(methyl methacrylate) (PMMA). When dipped into the Cr etchant, the exposed Cr layer was eliminated from the glass wafer in a short period of time. In order to ultimately fabricate the Ti/Au dot arrays, Ti and Au were deposited onto the arrays with a thickness of 2 and 40nm, respectively. The lift-off procedure was carried out in the Cr etchant using sonication in order to completely remove the residual Cr/PMMA layer. The fabricated Au nano-dot array was then immersed in an Ag enhancing solution and then into an ethanol solution containing (N-(6-(Biotinamido)hexyl)-3'-(2'-pyridyldithio)-propionamide (Biotin-HPDP). The substrate was analyzed using a correlated atomic force microscopy (AFM) and confocal Raman spectroscopy. Through this procedure, position-dependent surface-enhanced Raman spectroscopy (SERS) signals could be obtained.     

Verification of surfactant CHAPS effect using AFM for making biomemory device consisting of recombinant azurin monolayer

In this study, a protein-based biomemory device was developed using a surface modified recombinant azurin layer and its surface characteristics were analyzed by atomic force microscopy. The cysteine-modified azurin used for this purpose was a metalloprotein that had redox properties. To immobilize the metalloprotein on the Au substrates, the cysteine-modified azurin layer was self-assembled on the Au surface through a covalent bond between the thiol group on the cysteine and the Au surface. In our previous work, we showed that this protein layer was formed as cohesive clusters on Au surface through physical adsorption. To reduce the formation of these cohesion clusters, a zwitterionic surfactant, (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate) (CHAPS) was introduced to modify the surface properties. Using this approach, we found that CHAPS significantly reduced the amount of cysteine-modified azurin aggregates that nonspecifically adsorbed to the Au substrate. Atomic force microscopy was used to analyze the modified-surface. Based on this analysis, the size of the recombinant azurin clusters when CHAPS was used were about 15–25 nm whereas aggregates of 150–200 nm were observed in the absence of CHAPS. In addition, Raman spectroscopy was performed to confirm the retention of azurin molecules self-assembled on the Au surface. Electrochemical results using cyclic voltammetry indicated that recombinant azurin was successfully immobilized onto the Au surface with CHAPS and its redox property remained intact. Chronoamperometry was then used to demonstrate the memory characteristics of this azurin-based fabricated memory device. The combined results of this study show that CHAPS can significantly reduce the size of protein aggregates that become immobilized on the surface without a loss of the electrochemical properties of the protein.     

Characterization of biomolecule immobilization by scanning force microscopy using a wet-masking technique

The assemblage of molecular layers was investigated using scanning force microscopy (SFM). A wet-masking technique was used for the preparation of monolayer steps by partial masking of the substrate with an elastomeric mask during incubation in the modification buffers. The subsequent adsorption of biotin, streptavidin, and biotiny lated beads onto a gold substrate was investigated by SFM visualization of the created steps. The molecular layers were characterized based on measurements of step height and surface roughness. The simultaneous visualization of the surface before and after modification minimizes artifacts introduced by changes in tip shape or imaging parameters.     

Characterization of an ultraviolet irradiation chamber to monitor molecular photodegradation by Raman spectroscopy

A homemade ultraviolet chamber is reported to induce photochemical changes with characterization by Raman spectroscopy. The equipment has compartments for ultraviolet-A (UV-A) (8 lamps of 8 W) and for UV-A + ultraviolet-B (UV-B) irradiation (4 lamps of 26 W and 1 lamp of 15 W). The irradiance was measured 3, 5, and 10 cm from the light sources. The maximum irradiance was obtained at 3 cm (UV-A: 2.66 mW/cm² and UV-A + UV-B: 4.30 mW/cm²). The chamber internal temperature was stabilized at 30°C after 1 hr of operation with an internal relative humidity of approximately 45%. 10% Collagen was irradiated with UV-A at 2.0 mW/cm² for 3 hr with changes in Raman peaks at 1253, 1271, 1453, and 1660 cm^?1 indicating changes in conformation. 5% Atenolol was irradiated with UV-A + UV-B at 4.30 mW/cm² for 8 hours with changes to Raman peaks at 822, 1186, 1206, 1248, and 1618 cm^?1. A commercial insect repellent was irradiated with UV-A + UV-B at 4.30 mW/cm² for 8 hr and decreases in Raman intensity were observed at 526, 690, 1003, and 1606 cm^?1 due to degradation of N, N-diethyl-m-toluamide. The results demonstrate proper operation of the irradiation chamber with Raman spectroscopic monitoring.     

Returning to the same area of hair surfaces before and after treatment: a longitudinal AFM technique

We report the use of longitudinal (aspect ratio > 1 : 1) scanning atomic force microscopy as an aid in returning to the same area of hair fibres after bleaching, treatment with a commercial shampoo or the application of a ‘leave‐on’ conditioner product. The bleaching treatment used in this study was not found to affect the cuticular architecture and lateral force microscopy (LFM) also showed little difference after treatment, reflecting the homogeneity of the newly revealed surfaces. After treatment with a commercial shampoo, the hair sample again showed very little difference in topography or lateral force characteristics. Hair treated with the leave‐on conditioner product also showed no major topographical changes. LFM traces, however, showed regions between the ghost edge, marking the original position of the scale edge before cuticular erosion, and the existing scale edge, to have higher frictional properties than distal regions of the cuticle. A thin film of the leave‐on product thus seems to form in this region and extends from the foot of the scale edge.     

Effects of ultraviolet B radiation on physicochemical properties of human hair shaft

Changes in physicochemical properties of the human hair shaft induced by UVB during the summer climate were investigated. Using atomic force microscopy and thermogravimetric analyzer measurements, we investigated the changes in hair shaft physical properties including morphology, roughness, and water content. The changes in chemical properties were investigated by using FTIR spectroscopy. Hair samples from 20 females without any chemical or physical stresses were investigated. The samples were exposed to various UVB doses of 400, 2,000, and 4,000 mJ/cm(2), which correspond to the radiation times of 0.44, 2.22, and 4.44 h, respectively. The morphology and surface of the hair shaft was not significantly affected by UVB irradiation. The components of keratin were affected by the UVB irradiation where the amide A bands at 3,263 and 3,246 cm(-1) of the proximal and distal roots, respectively, were shifted to higher band region because α-helix converted to β-sheet, and the stretching modes of C-H lipid alkyl chains at 2,965 and 2,850 cm(-1) in the proximal and distal roots, respectively, were developed. As the UVB intensity increased, the water content of the proximal root decreased.     

STM and AFM investigations of high-Tc superconductors

We have studied the (001) surface of single crystal YBa₂Cu₃O_7-x high-T_c superconductors using scanning tunnelling microscopy (STM) and atomic force microscopy (AFM) at room temperature at ambient pressure. Both methods show flat terraces with steps which are multiples of the c-axis lattice constant (of 1·17 nm) high. Our results show that the bulk crystal structure extends to the surface and that the crystals were formed by island growth. Only occasionally tunnelling was possible with sample bias voltages below +1·0 V. We interpret the observed voltage dependence and the difficulty to get good STM images to be due to the presence of a less-conducting surface layer. Auger spectroscopy indicates that carbon is present at the surface, which is probably related to a contamination layer.     

Nanostructures in silicon investigated by atomic force microscopy and surface photovoltage spectroscopy

Surface photovoltage spectroscopy (SPS) and conductive atomic force microscopy (C-AFM) have been used for the characterization of nanocrystalline hydrogenated Si (nc-Si:H). This is a promising material both for silicon-based opto-electronics as well as for photovoltaic applications. Notwithstanding its interesting properties many issues regarding the material electronic and optical properties are not completely understood. The present contribution reports microscopic and spectroscopic analyses of nc-Si:H films grown for photovoltaic applications by low-energy plasma-enhanced chemical vapor deposition technique. Electronic levels associated with defect states were investigated by SPS, whereas the conduction mechanism at a microscopic level was investigated by C-AFM.     

Large-area topography analysis and near-field Raman spectroscopy using bent fibre probes

We present a method for combined far‐field Raman imaging, topography analysis and near‐field spectroscopy. Surface‐enhanced Raman spectra of Rhodamine 6G (R6G) deposited on silver nanoparticles were recorded using a bent fibre aperture‐type near‐field scanning optical microscope (NSOM) operated in illumination mode. Special measures were taken to enable optical normal‐force detection for control of the tip–sample distance. Comparisons between far‐field Raman images of R6G‐covered Ag particle aggregates with topographic images recorded using atomic force microscopy (AFM) indicate saturation effects due to resonance excitation.     

Imaging and strain analysis of nano-scale SiGe structures by tip-enhanced Raman spectroscopy

The spatial resolution and high sensitivity of tip-enhanced Raman spectroscopy allows the characterization of surface features on a nano-scale. This technique is used to visualize silicon-based structures, which are similar in width to the transistor channels in present leading-edge CMOS devices. The reduction of the intensive far-field background signal is crucial for detecting the weak near-field contributions and requires beside a careful alignment of laser polarization and tip axis also the consideration of the crystalline sample orientation. Despite the chemical identity of the investigated sample surface, the structures can be visualized by the shift of the Raman peak positions due to the patterning induced change of the stress distribution within lines and substrate layer. From the measured peak positions the intrinsic stress within the lines is calculated and compared with results obtained by finite element modeling. The results demonstrate the capability of the tip-enhanced Raman technique for strain analysis on a sub-50 nm scale.     

Combined ultramicrotomy for AFM and TEM using a novel sample holder

A new sample holder that allows combined microtomy for atomic force microscopy (AFM) and transmission electron microscopy (TEM) is described. The main feature of this sample holder is a small central part holding the sample. This central part fits into the head of an atomic force microscope. AFM measurements can be performed with a sample mounted in this central part of the sample holder. This makes the alignment of a microtomed bulk sample unnecessary, and offers the opportunity of an easy and fast combined sample preparation for AFM and TEM.