Evaporative Drying Induced Self-Assembly of Epicuticular Wax: A Biomimetic Approach in Tuning Surface Roughness.

Epicuticular wax is an example of a naturally created functional material that forms a layer on the outermost surface of plants with the objective to protect them from adverse environmental conditions, such as UV-solar radiation, uncontrolled water loss, microbial attacks, and so forth. Their functionalities are often attributed to the chemical composition of the wax as well as the physical structuration formed by the wax crystals on the surface. With this work, we present a simple, one-step biomimetic approach to replicate similar surface structures, on model substrate, using wax extracted from Euphorbia Cerifera (Candelilla wax). First, we describe formation of structured wax due to self-assembly induced by evaporative drying on quartz plates. Subsequently, we highlight the fundamental physical parameters required to tune the surface morphology. Our experiments reveal that it is possible to achieve considerably diverse surface morphologies depending on the solvent properties and deposition temperature. This diversity is due to the kinetics of recrystallization of wax during evaporation of solvent which, in turn, is primarily driven by the solubility of wax as well as evaporation rate of the solvent. Thus, the final morphology that we obtain is an interplay between recrystallization kinetics and solvent evaporation. Additionally, the degree of crystallinity of the structured films could also be tuned by solvent polarity. Surprisingly, X-ray diffraction indicates that the crystalline structure at the molecular level remains similar to that of bulk Candelilla wax. Our results provide fundamental insights into the replication of epicuticular wax films and identification of tuning parameters to obtain different surface morphologies with the same wax material for potential bioinspired multifunctional coatings in cosmetic applications.

Sensory neuron activation from topical treatments modulates the sensorial perception of human skin.

Neural signaling of skin sensory perception from topical treatments is often reported in subjective terms such as a sensation of skin "tightness" after using a cleanser or "softness" after applying a moisturizer. However, the mechanism whereby cutaneous mechanoreceptors and corresponding sensory neurons are activated giving rise to these perceptions has not been established. Here, we provide a quantitative approach that couples in vitro biomechanical testing and detailed computational neural stimulation modeling along with a comprehensive in vivo self-assessment survey to demonstrate how cutaneous biomechanical changes in response to treatments are involved in the sensorial perception of the human skin. Strong correlations are identified between reported perception up to 12 hours post treatment and changes in the computed neural stimulation from mechanoreceptors residing deep under the skin surface. The study reveals a quantitative framework for understanding the biomechanical neural activation mechanism and the subjective perception by individuals.

Poroelastic behavior and water permeability of human skin at the nanoscale.

Topical skin care products and hydrating compositions (moisturizers or injectable fillers) have been used for years to improve the appearance of, for example facial wrinkles, or to increase "plumpness". Most of the studies have addressed these changes based on the overall mechanical changes associated with an increase in hydration state. However, little is known about the water mobility contribution to these changes as well as the consequences to the specific skin layers. This is important as the biophysical properties and the biochemical composition of normal stratum corneum, epithelium, and dermis vary tremendously from one another. Our current studies and results reported here have focused on a novel approach (dynamic atomic force microscopy-based nanoindentation) to quantify biophysical characteristics of individual layers of ex vivo human skin. We have discovered that our new methods are highly sensitive to the mechanical properties of individual skin layers, as well as their hydration properties. Furthermore, our methods can assess the ability of these individual layers to respond to both compressive and shear deformations. In addition, since human skin is mechanically loaded over a wide range of deformation rates (frequencies), we studied the biophysical properties of skin over a wide frequency range. The poroelasticity model used helps to quantify the hydraulic permeability of the skin layers, providing an innovative method to evaluate and interpret the impact of hydrating compositions on water mobility of these different skin layers.

Dilution-Induced Deposition of Concentrated Binary Mixtures of Cationic Polysaccharides and Surfactants.

This work investigates the effect of dilution on the phase separation process of binary charged polysaccharide-surfactant mixtures formed by two cationic polysaccharides and up to four surfactants of different nature (anionic, zwitterionic, and neutral), as well as the potential impact of dilution-induced phase separation on the formation of conditioning deposits on charged surfaces, mimicking the negative charge and wettability of damaged hair fibers. The results obtained showed that the dilution behavior of model washing formulations (concentrated polysaccharide-surfactant mixtures) cannot be described in terms of a classical complex precipitation framework, as phase separation phenomena occur even when the aggregates are far from the equilibrium phase separation composition. Therefore, dilution-enhanced deposition cannot be predicted in terms of the worsening of colloidal stability due to the charge neutralization phenomena, as common phase separation and, hence, enhanced deposition occurs even for highly charged complexes.

Shearing friction behaviour of synthetic polymers compared to a functionalized polysaccharide on biomimetic surfaces: models for the prediction of performance of eco-designed formulations.

The substitution of natural, bio-based and/or biodegradable polymers for those of petrochemical origin in consumer formulations has become an active area of research and development as the sourcing and destiny of material components becomes a more critical factor in product design. These polymers often differ from their petroleum-based counterparts in topology, raw material composition and solution behaviour. Effective and efficient reformulation that maintains comparable cosmetic performance to existing products requires a deep understanding of the differences in frictional behaviour between polymers as a function of their molecular structure. In this work, we simulate the tribological behaviour of three topologically distinct polymers in solution with surfactants and in contact with hair-biomimetic patterned surfaces. We compare a generic functionalized polysaccharide to two performant polymers used in shampoo formulations: a strongly positively charged polyelectrolyte and a zwitterionic copolymer. Topological differences are expected to affect rheological properties, as well as their direct interaction with structured biological substrates. Using a refined Martini-style coarse-grained model we describe the polymer-dependent differences in aggregation behaviour as well as selective interactions with a biomimetic model hair surface. Additionally, we introduce a formalism to characterize the response of the solution to shear as an initial study on lubrication properties, which define the sensorial performance of these systems in cosmetics (i.e., manageability, touch, etc.). The tools and techniques presented in this work illustrate the strength of molecular simulation in eco-design of formulation as a complement to experiment. These efforts help advance our understanding of how we can relate complex atomic-scale solution behaviour to relevant macroscopic properties. We expect these techniques to play an increasingly important role in advancing strategies for green polymer formulation design by providing an understanding for how new polymers could reach and even exceed the level of performance of existing polymers.

3D Molecular Imaging of Stratum Corneum by Mass Spectrometry Suggests Distinct Distribution of Cholesteryl Esters Compared to Other Skin Lipids.

The crucial barrier properties of the stratum corneum (SC) depend critically on the design and integrity of its layered molecular structure. However, analysis methods capable of spatially resolved molecular characterization of the SC are scarce and fraught with severe limitations, e.g., regarding molecular specificity or spatial resolution. Here, we used 3D time-of-flight secondary ion mass spectrometry to characterize the spatial distribution of skin lipids in corneocyte multilayer squams obtained by tape stripping. Depth profiles of specific skin lipids display an oscillatory behavior that is consistent with successive monitoring of individual lipid and corneocyte layers of the SC structure. Whereas the most common skin lipids, i.e., ceramides, C24:0 and C26:0 fatty acids and cholesteryl sulfate, are similarly organized, a distinct 3D distribution was observed for cholesteryl oleate, suggesting a different localization of cholesteryl esters compared to the lipid matrix separating the corneocyte layers. The possibility to monitor the composition and spatial distribution of endogenous lipids as well as active drug and cosmetic substances in individual lipid and corneocyte layers has the potential to provide important contributions to the basic understanding of barrier function and penetration in the SC.

From decoding the perception of tightness to a clinical proof of soothing effects derived from natural ingredients in a moisturizer.

OBJECTIVE: To decode the feeling of skin tightness after application of a cosmetic product and how to soothe this discomfort. To pursue this aim, we considered the ingredient's effect on stratum corneum (SC) biomechanics to differentiate between consumers prone to tightness from those that are not and correlate these effects with mechanoreceptor activation. METHODS: In vivo clinical trials were used to assess the tightness perception dichotomy between groups of Caucasian women; in vitro experiments were used to measure the mechanical stresses induced in the SC after cleanser and moisturizer application; and in silico simulations were used to illustrate how the measured mechanical stresses in the SC result in the development of strains at the depth of cutaneous mechanoreceptors, triggering tightness perceptual responses. RESULTS: Before any cream application, women prone to tightness tend to have a more rigid SC than their less sensitive counterparts, however cleanser application increases SC stiffness in all women. Surprisingly, no correlation was found between tightness perception and hydration measurements by the Corneometer or barrier function, as evaluated by transepidermal water loss. Self-declared tightness and dryness scores were strongly associated with a self-described sensitive skin. After application of the optimized moisturizing formula, Osmoskin® containing natural waxes with good filming properties, consumers report a strong decrease in tightness and dryness perception. These results match with laboratory experiments where the cleanser was shown to increase SC drying stresses by 34%, while subsequent application of Osmoskin® decreased stresses by 48%. Finite element modelling, using experimental results as input, elucidates the differences in perception between the two groups of women. It makes clear that Osmoskin® changes the mechanical status of the SC, producing strains in underlying epidermis that activates multiple cutaneous mechano-receptors at a level correlated with the self-perceived comfort. CONCLUSION: Integration of the in vivo, in vitro and in silico approaches provides a novel framework for fully understanding how skin tightness sensations form and propagate, and how these sensations can be alleviated through the design of an optimized moisturizer.

OBJECTIF: Décoder l'impression de tiraillement de la peau après l'application d'un produit cosmétique et la manière d'apaiser cette sensation désagréable. Pour poursuivre cet objectif, nous avons pris en compte l'effet de l'ingrédient sur la biomécanique de la couche cornée afin de différencier les consommatrices sujettes à un tiraillement de celles qui ne le sont pas et de corréler ces effets avec l'activation des mécanorécepteurs. MÉTHODES: Des essais cliniques in vivo ont été utilisés pour évaluer la dichotomie de perception de tiraillement entre des groupes de femmes de race caucasienne; des expériences in vitro ont été utilisées pour mesurer les contraintes mécaniques induites dans la couche cornée après application d'un produit nettoyant et d'un produit hydratant; et des simulations in silico ont servi à illustrer comment les contraintes mécaniques mesurées dans la couche cornée entraînent le développement de souches à la profondeur des mécanorécepteurs cutanés, qui déclenchent les réponses perceptives de tiraillement. RÉSULTATS: Avant toute application de crème, les femmes sujettes au tiraillement tendent à avoir une couche cornée plus rigide que leurs homologues moins sensibles, mais l'application d'un produit nettoyant augmente la raideur de la couche cornée chez toutes les femmes. Étonnamment, aucune corrélation n'a été observée entre la perception de tiraillement et les mesures d'hydratation réalisées par le cornéomètre ou la fonction barrière, évaluée par la perte d'eau transépidermique. Les scores de tiraillement et de sécheresse auto-déclarés étaient fortement corrélés à une peau décrite par les sujets elles-mêmes comme sensible. Après application de la formule hydratante optimisée, Osmoskin®, qui contient des cires naturelles ayant de bonnes propriétés de dépôt de film, les consommateurs rapportent une forte diminution de la sensation de tiraillement et de sécheresse. Ces résultats concordent avec les expériences en laboratoire où le produit nettoyant s'est avéré augmenter les contraintes de séchage de la couche cornée de 34 %, tandis que l'application ultérieure d'Osmoskin® a réduit les contraintes de 48 %. La modélisation à éléments finis, en utilisant les résultats expérimentaux comme données, élucide les différences de perception entre les deux groupes de femmes. Il est clair qu'Osmoskin® modifie l'état mécanique de la couche cornée, et produit des souches dans l'épiderme sous-jacent qui activent plusieurs mécano-récepteurs cutanés à un niveau corrélé au confort perçu par la patiente. CONCLUSION: La combinaison des approches in vivo, in vitro et in silico fournit un nouveau cadre pour comprendre pleinement comment les sensations de tiraillement de la peau se forment et se propagent, et comment elles peuvent être soulagées en mettant au point une crème hydratante optimisée.

Study of the Dilution-Induced Deposition of Concentrated Mixtures of Polyelectrolytes and Surfactants.

Mixtures of polyelectrolytes and surfactants are commonly used in many technological applications where the challenge is to provide well-defined modifications of the surface properties, as is the case of washing formulations in cosmetics. However, if contemporary experimental and theoretical methods can provide insights on their behavior in concentrated formulations, less is known on their behavior under practical use conditions, e.g., under dilution and vectorization of deposits. This makes it difficult to make predictions for specific performance, as, for example, good hair manageability after a shampoo or a comfortable sensorial appreciation after a skin cleanser. This is especially important when considering the formulation of new, more eco-friendly formulations. In this work, a detailed study of the phase separation process induced by dilution is described, as well as the impact on the deposition of conditioning material on negatively charged surfaces. In order to gain a more detailed physical insight, several polyelectrolyte-surfactant pairs, formed by two different polymers and five surfactants that, although non-natural or eco-friendly, can be considered as models of classical formulations, have been studied. The results evidenced that upon dilution the behavior, and hence its deposition onto the surface, cannot be predicted in terms of the behavior of simpler pseudo-binary (mixtures of a polymer and a surfactant) or pseudo-ternary mixtures (two polymers and a surfactant). In many cases, phase separation was observed for concentrations similar to those corresponding to the components in some technological formulations, whereas the latter appeared as monophasic systems. Therefore, it may be assumed that the behavior in multicomponent formulations is the result of a complex interplay of synergistic interactions between the different components that will require revisiting when new, more eco-sustainable ingredients are considered.

Adsorption and Desorption of Polymers on Bioinspired Chemically Structured Substrates.

Natural biological surfaces exhibit interesting properties due to their inhomogeneous chemical and physical structure at the micro- and nanoscale. In the case of hair or skin, this also influences how waterborne macromolecules ingredients will adsorb and form cosmetically performing deposits (i.e., shampoos, cleansers, etc.). Here, we study the adsorption of hydrophilic flexible homopolymers on heterogeneous, chemically patterned substrates that represent the surface of the hair by employing coarse-grained molecular dynamics simulations. We develop a method in which the experimental images of the substrate are used to obtain information about the surface properties. We investigate the polymer adsorption as a function of polymer chain length and polymer concentration spanning both dilute and semidilute regimes. Adsorbed structures are quantified in terms of trains, loops, and tails. We show that upon increasing polymer concentration, the length of tails and loops increases at the cost of monomers belonging to trains. Furthermore, using an effective description, we probe the stability of the resulting adsorbed structures under a linear shear flow. Our work is a first step toward developing models of complex macromolecules interacting with realistic biological surfaces, as needed for the development of more ecofriendly industrial products.

A Curly Q: Is Frizz a Matter of Friction?

The oft discussed and fretted over environmental influences on hair have led to a popular consensus which suggests that elevated temperature and humidity lead to frizzier, wilder hair. However, few attempts at actually quantifying these effects have been made. Although frizziness is usually perceived visually, here the influence of variations in temperature and humidity on the tactile perception and friction of curly and straight hair were investigated. It is shown that changes in humidity may disproportionately affect perceived frizziness of curly hair by touch due to concurrent changes in the tactile friction.

Surface science of cosmetic substrates, cleansing actives and formulations.

The development of shampoo and cleansing formulations in cosmetics is at a crossroads due to consumer demands for better performing, more natural products and also the strong commitment of cosmetic companies to improve the sustainability of cosmetic products. In order to go beyond traditional formulations, it is of great importance to clearly establish the science behind cleansing technologies and appreciate the specificity of cleansing biological surfaces such as hair and skin. In this review, we present recent advances in our knowledge of the physicochemical properties of the hair surface from both an experimental and a theoretical point of view. We discuss the opportunities and challenges that newer, sustainable formulations bring compared to petroleum-based ingredients. The inevitable evolution towards more bio-based, eco-friendly ingredients and sustainable formulations requires a complete rethink of many well-known physicochemical principles. The pivotal role of digital sciences and modelling in the understanding and conception of new ingredients and formulations is discussed. We describe recent numerical approaches that take into account the specificities of the hair surface in terms of structuration, different methods that study the adsorption of formulation ingredients and finally the success of new data-driven approaches. We conclude with practical examples on current formulation efforts incorporating bio-surfactants, controlling foaming and searching for new rheological properties.

Tribology of an assembly of hairs: Influence of multiscale surface chemistry and structure on sensorial tactile properties.

BACKGROUND: Hair fibers may be either oriented in a common direction or randomly arranged. Fiber arrangement as well as cosmetic treatment control the sensorial perception. The present study explores the respective influence of these two aspects by predicting the product performance in terms of tactile perception. MATERIALS AND METHODS: Friction forces between hair swatches of different curl patterns using a finger-like probe have been measured to better mimic real-life hair/finger contact. Measurements of fiber alignment, hair diameter (thickness), and compression tests were performed on natural and treated swatches to assess the respective weight of these parameters. RESULTS: Conditioned hair exhibit an adhesive behavior measured at the start of the frictional movement. Conversely, natural hair is influenced by fiber reorientation. After a few seconds, friction-related signals stabilize. Thus, the averaged friction forces do not only depend on hair thickness, but increase with a decreased alignment of the fibers. CONCLUSIONS: Intrinsic (diameter/curliness) and external (orientation/ friction/compression) characteristics allow to define a model of "macroscopic" roughness linked to the sensorial characterization. As friction of hair swatches depends upon fiber alignment and coating, this combined approach is potentially a very useful in vitro test, as an alternative or complementary method to sensory tests.

Turning autophobic wetting on biomimetic surfaces into complete wetting by wetting additives.

Autophobicity or pseudo partial wetting, a phenomenon of a liquid not spreading on its own monolayer, is characterized by an energy barrier that prevents the growth of a wetting film beyond the monolayer thickness. Applying a molecularly detailed self-consistent field theory we illustrate how autophobic wetting can be overcome by wetting additives. More specifically we use an emulsifier which keeps the interfacial tension between the wetting component and the majority solvent low, and a co-solvent additive which partitions inside the film and then destroys the molecular order in it so that the barrier for film growth is cleared. An application wherein it is believed that autophobic wetting is counteracted by such a set of wetting additives is found in an antidandruff shampoo formulation. We have experimental results that show thick deposits onto hydrophobic hair surfaces by administration of the antidandruff shampoo. The complementary modeling of such a system suggests that the active ingredient plays the role of the co-solvent additive. As significant amounts of the co-solvent additives are needed to approach the completely wet state, the formulation naturally brings large amounts of active ingredient to the root of the hair where its presence is required.

Imaging and quantifying drug delivery in skin - Part 2: Fluorescence andvibrational spectroscopic imaging methods.

Understanding the delivery and diffusion of topically-applied drugs on human skin is of paramount importance in both pharmaceutical and cosmetics research. This information is critical in early stages of drug development and allows the identification of the most promising ingredients delivered at optimal concentrations to their target skin compartments. Different skin imaging methods, invasive and non-invasive, are available to characterize and quantify the spatiotemporal distribution of a drug within ex vivo and in vivo human skin. The first part of this review detailed invasive imaging methods (autoradiography, MALDI and SIMS). This second part reviews non-invasive imaging methods that can be applied in vivo: i) fluorescence (conventional, confocal, and multiphoton) and second harmonic generation microscopies and ii) vibrational spectroscopic imaging methods (infrared, confocal Raman, and coherent Raman scattering microscopies). Finally, a flow chart for the selection of imaging methods is presented to guide human skin ex vivo and in vivo drug delivery studies.

Imaging and quantifying drug delivery in skin - Part 1: Autoradiography and mass spectrometry imaging.

In this two-part review we present an up-to-date description of different imaging methods available to map the localization of drugs on skin as a complement of established ex-vivo absorption studies. This first part deals with invasive methods which are grouped in two classes according to their underlying principles: i) methods using radioactivity such as autoradiography and ii) mass spectrometry methods such as MALDI and SIMS. For each method, a description of the principle is given along with example applications of imaging and quantifying drug delivery in human skin. Thanks to these techniques a better assessment of the fate of drugs is obtained: its localization on a particular skin structure, its potential accumulation, etc. A critical comparison in terms of capabilities, sensitivity and practical applicability is included that will help the reader to select the most appropriate technique depending on the particular problem to be solved.

Effect of molecular structure of eco-friendly glycolipid biosurfactants on the adsorption of hair-care conditioning polymers.

Pseudo-binary mixtures of different glycolipids, four different rhamnolipids (RL) and an alkyl polyglucoside (APG), with poly(diallyl-dimethylammonium chloride) (PDADMAC) have been studied in relation to their adsorption onto negatively charged surfaces to shed light on the impact of the molecular structure of surfactants from natural sources (instead of synthetic surfactant, such as sodium laureth sulfate) on the adsorption of hair-conditioning polymers. For this purpose, the self-assembly of such mixtures in aqueous solution and their adsorption onto negatively charged surfaces mimicking the negative charge of damaged hair fibres have been studied combining experiments and self-consistent field (SCF) calculations. The results show that the specific physico-chemical properties of the surfactants (charge, number of sugar rings present in surfactant structure and length of the hydrocarbon length) play a main role in the control of the adsorption process, with the adsorption efficiency and hydration being improved in relation to conventional sulfate-based systems for mixtures of PDADMAC and glycolipids with the shortest alkyl chains. SCF calculations and Energy Dispersive X-Ray Spectroscopy (EDS) analysis on real hair confirmed such observations. The results allow one to assume that the characteristic of the surfactants, especially rhamnolipids, conditions positively the adsorption potential of polyelectrolytes in these model systems. This study provides important insights on the mechanisms underlying the performance of more complex but eco-friendly washing formulations.

Effects of Imprinted 3D Surface Patterning on Localized Changes in the Tribology of Human Stratum Corneum.

Natural surfaces may exhibit remarkable surface properties due to their structure. In the case of skin, its surface topography (microrelief) influences many of its perceived sensorial properties (shine, color, touch). Imprinted patterns can modify the original microrelief, inducing a completely new set of perceived properties. To explore the effects of superimposed biomimetic surface textures on the friction of skin, human stratum corneum was prepared with and without an imprinted regular, micrometer-sized, 3D grid pattern. Atomic Force Microscopy (AFM) and optical profilometry indicated that the inherent, smaller-scale roughness of the stratum corneum remained when lines with heights of 20-200 µm and spacings of 600-2000 µm were introduced, but it was somewhat reduced on the grid lines. Surface Forces Apparatus (SFA) friction experiments on stratum corneum were performed at low speed (µm/s, back-and-forth sliding) and at more realistic, high speed (cm/s, rotational sliding). Two stratum corneum surfaces in contact did not adhere to one another, and they had a friction coefficient µ of 0.1, or lower, at low sliding speed. An interesting loading-unloading hysteresis was observed, with lower friction force on unloading, in particular, when the contact was on a grid line of the patterned samples. This suggests that the patterning locally induced different mechanical properties of the stratum corneum and that its recovery was not immediate on unloading. When one stratum corneum surface slid against a rigid glass surface, the friction coefficient was always higher than that when two stratum corneum surfaces were in contact. At high sliding speed, much higher friction coefficients were found between one stratum corneum surface and a rigid, smooth surface, µ ≥ 1. The results demonstrate that topograpic patterning by imprinting clearly modifies the tribological response of stratum corneum. This approach provides a simple method for exploring the development of biomimetic modifications of skin texture.

Anisotropic cellular forces support mechanical integrity of the Stratum Corneum barrier.

The protective function of biological surfaces that are exposed to the exterior of living organisms is the result of a complex arrangement and interaction of cellular components. This is the case for the most external cornified layer of skin, the stratum corneum (SC). This layer is made of corneocytes, the elementary 'flat bricks' that are held together through adhesive junctions. Despite the well-known protective role of the SC under high mechanical stresses and rapid cell turnover, the subtleties regarding the adhesion and mechanical interaction among the individual corneocytes are still poorly known. Here, we explore the adhesion of single corneocytes at different depths of the SC, by pulling them using glass microcantilevers, and measuring their detachment forces. We measured their interplanar adhesion between SC layers, and their peripheral adhesion among cells within a SC layer. Both adhesions increased considerably with depth. At the SC surface, with respect to adhesion, the corneocyte population exhibited a strong heterogeneity, where detachment forces differed by more than one order of magnitude for corneocytes located side by side. The measured detachment forces indicated that in the upper-middle layers of SC, the peripheral adhesion was stronger than the interplanar one. We conclude that the stronger peripheral adhesion of corneocytes in the SC favors an efficient barrier which would be able to resist strong stresses.

Imaging the distribution of skin lipids and topically applied compounds in human skin using mass spectrometry.

The barrier functions of skin against water loss, microbial invasion and penetration of xenobiotics rely, in part, on the spatial distribution of the biomolecular constituents in the skin structure, particularly its horny layer (stratum corneum). However, all skin layers are important to describe normal and dysfunctional skin conditions, and to develop adapted therapies or skin care products. In this work, time-of-flight secondary ion mass spectrometry (ToF-SIMS) combined with scanning electron microscopy (SEM) was used to image the spatial distribution of a variety of molecular species, from stratum corneum down to dermis, in cross-section samples of human abdominal skin. The results demonstrate the expected localization of ceramide and saturated long-chain fatty acids in stratum corneum (SC) and cholesterol sulfate in the upper part of the viable epidermis. The localization of exogenous compounds is demonstrated by the detection and imaging of carvacrol (a constituent of oregano or thyme essential oil) and ceramide, after topical application onto ex vivo human skin. Carvacrol showed pronounced accumulation to triglyceride-containing structures in the deeper parts of dermis. In contrast, the exogenous ceramide was found to be localized in SC. Furthermore, the complementary character of this approach with classical ex vivo skin absorption analysis methods is demonstrated.