Ana-Maria Pena

Ana-Maria Pena
L'Oréal · Research and Innovation, Physics Department

Doctor of Philosophy

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63
Publications
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1,837
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Introduction
Skills and Expertise

Publications

Publications (63)
Article
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Characterizing melanins in situ and determining their 3D z-epidermal distribution is paramount for understanding physiological/pathological processes of melanin neosynthesis, transfer, degradation or modulation with external UV exposure or cosmetic/pharmaceutical products. Multiphoton fluorescence intensity- and lifetime-based approaches have been...
Article
Full-text available
Solar ultraviolet longwave UVA1 exposure of human skin has short-term consequences at cellular and molecular level, leading at long-term to photoaging. Following exposure, reactive oxygen species (ROS) are generated, inducing oxidative stress that might impair cellular metabolic activity. However, the dynamic of UVA1 impact on cellular metabolism r...
Article
Full-text available
The cover image is based on the Original Article In vivo multiphoton imaging for non‐invasive time course assessment of retinoids effects on human skin by Emmanuelle Tancrède‐Bohin et al., https://doi.org/10.1111/srt.12877.
Article
Full-text available
Background In vivo multiphoton imaging and automatic 3D image processing tools provide quantitative information on human skin constituents. These multiphoton‐based tools allowed evidencing retinoids epidermal effects in the occlusive patch test protocol developed for antiaging products screening. This study aimed at investigating their relevance fo...
Article
Full-text available
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 compartm...
Article
In this two parts review we present an up-do-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 and are grouped in two classes according to their underlying principles: i) methods using radioactivity s...
Article
Full-text available
Two-photon imaging of endogenous fluorescence can provide physiological and metabolic information from intact tissues. However, simultaneous imaging of multiple intrinsic fluorophores, such as nicotinamide adenine dinucleotide(phosphate) (NAD(P)H), flavin adenine dinucleotide (FAD) and retinoids in living systems is generally hampered by sequential...
Article
Objective: The roots of the herb Paeonia lactiflora ("White Peony") are used in association with other herbs in traditional clinical cosmetic practice in China as oral treatment for skin pigmentary disorders, such as brown or dark pigmentary spots. However, the skin depigmenting potential of Paeonia lactiflora root extract and its main ingredient...
Article
Background The occlusive patch test developed for assessing topical retinoids activity in human skin has been extended as a short-term screening protocol for anti-ageing agents. In this model, biopsies are performed at the end of the occlusion period for morphological and immuno-histochemistry analysis. Multiphoton microscopy is a recent non-invasi...
Conference Paper
Up to now, fast in situ 3D non-invasive melanin detection is a still challenging objective. In the past years, different methods based on time-resolved two-photon excited fluorescence were proposed. These methods take advantage of the specific melanin fluorescence lifetime properties and use multiphoton FLIM imaging (Fluorescence-Lifetime Imaging M...
Conference Paper
Melanocyte-keratinocyte co-culture models are useful in vitro systems for identifying the de-pigmenting or pro-pigmenting potential of a given cosmetic ingredient. This potential can be estimated by calculating the density of melanin present within this model. Multiphoton microscopy is a privileged microscopy technique for such evaluation, thanks t...
Article
In this paper, a method to segment elongated objects is proposed. It is based on attribute profiles and area stability. Images are represented as component trees using a threshold decomposition. Then, some attributes are computed on each node of the tree. Finally, the attribute profile is analyzed to identify important events useful for segmentatio...
Article
BACKGROUND/PURPOSE: Multiphoton microscopy has emerged in the past decade as a useful noninvasive imaging technique for in vivo human skin characterization. However, it has not been used until now in evaluation clinical trials, mainly because of the lack of specific image processing tools that would allow the investigator to extract pertinent quant...
Article
Full-text available
Introduction It is now recognized that to adequately protect skin from sun damage, sunscreens require a photostable combination of ultraviolet (UV) filters with a suitable level of UVA protection. The total amount of UV filters should be as low as possible to avoid adverse skin reactions, potential environmental impact, and to ensure acceptable tex...
Conference Paper
Multiphoton microscopy has emerged in the past decade as a promising non-invasive skin imaging technique. The aim of this study was to assess whether multiphoton microscopy coupled to specific 3D image processing tools could provide new insights into the organization of different skin components and their age-related changes. For that purpose, we p...
Article
Multiphoton microscopy has emerged in the past decade as a promising tool for noninvasive skin imaging. Our aim was to evaluate the potential of multiphoton microscopy to detect topical corticosteroids side effects within the epidermis and to provide new insights into their dynamics. Healthy volunteers were topically treated with clobetasol propion...
Article
Full-text available
An optimized system for fast, high-resolution spectral imaging of in vivo human skin is developed and evaluated. The spectrograph is composed of a dispersive prism in combination with an electron multiplying CCD camera. Spectra of autofluorescence and second harmonic generation (SHG) are acquired at a rate of 8 kHz and spectral images within second...
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Dermal fibroblasts are responsible for the generation of mechanical forces within their surrounding extracellular matrix and can be potentially targeted by anti-aging ingredients. Investigation of the modulation of fibroblast contraction by these ingredients requires the implementation of three-dimensional in situ imaging methodologies. We use mult...
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We compare second harmonic generation (SHG) to histological and immunohistochemical techniques for the visualization and scoring of collagen in biological tissues. We show that SHG microscopy is highly specific for fibrillar collagens and that combined SHG and two-photon excited fluorescence (2PEF) imaging can provide simultaneous three-dimensional...
Article
We used intrinsic Second Harmonic Generation (SHG) by fibrillar collagen to visualize the three-dimensional architecture of collagen fibrosis at the micrometer scale using laser scanning nonlinear microscopy. We showed that SHG signals are highly specific to fibrillar collagen and provide a sensitive probe of the micrometer-scale structural organiz...
Article
The organization of collagen during fibrotic processes is poorly characterized because of the lack of appropriate methodologies. Here we show that multimodal multiphoton microscopy provides novel insights into lung fibrosis. We characterize normal and fibrotic pulmonary tissue in the bleomycin model, and show that second-harmonic generation by fibr...
Article
Full-text available
One principal advantage of multiphoton excitation microscopy is that it preserves its three-dimensional micrometer resolution when imaging inside light-scattering samples. For that reason two-photon-excited fluorescence microscopy has become an invaluable tool for cellular imaging in intact tissue, with applications in many fields of physiology. Th...
Article
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La microscopie multiphoton est une technique d'imagerie optique dont une des caractéristiques les plus remarquables est de fournir une information micrométrique en profondeur dans les tissus intacts. Un autre intérêt est la possibilité de visualiser la structure d'une cellule ou d'un tissu en utilisant des sources de contraste endogènes qui permett...
Article
We recorded multiphoton images of human skin biopsies using endogenous sources of nonlinear optical signals. We detected simultaneously two-photon excited fluorescence (2PEF) from intrinsic fluorophores and second harmonic generation (SHG) from collagen. We observed SHG from fibrillar collagens in the dermis, whereas no SHG was detectable from the...
Article
Full-text available
Lipid bodies have an important role in energy storage and lipid regulation. Here we show that lipid bodies are a major source of contrast in third-harmonic generation (THG) microscopy of cells and tissues. In hepatocytes, micrometer-sized lipid bodies produce a THG signal 1-2 orders of magnitude larger than other structures, which allows one to ima...
Article
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La fibrose est une réponse adaptative pathologique qui détruit non spécifiquement les tissus. Il s'agit d'un processus universel de réparation des tissus qui survient en réaction à de nombreux types d'agressions telles les contraintes mécaniques, les brûlures, les radiations ionisantes, l'ischémie, l'inflammation. Ces agressions concourent de maniè...
Article
An emerging application of multiphoton microscopy is the observation of unstained intact biological tissues based on intrinsic sources of nonlinear signals. However, reliable biomedical imaging requires a thorough analysis of these endogenous signals. Looking at skin biopsies, we focused on 2-Photon Excited Fluorescence (2PEF) arising from cytokera...
Article
We characterize the application of multiphoton microscopy to the observation of the extracellular matrix of fresh unstained vessels. Combined two-photon-excited fluorescence (2PEF) and second harmonic generation (SHG) imaging of large arteries reveals the architecture of elastin and collagen fibers in the vessel wall with remarkable specificity. We...
Article
Full-text available
We recorded one-photon excited fluorescence (1PEF) and two-photon excited fluorescence (2PEF) spectra of purified keratin from human epidermis, and determined the action cross section of this endogenous chromophore. We used this spectroscopic analysis to analyse multiphoton images of skin biopsies and assign the intrinsic fluorescence signals in th...
Article
We performed polarization-resolved surface second harmonic generation (SHG) experiments on thin films of collagen I and IV molecules, as well as conventional CD measurements. We found that collagen IV presents little CD and no SHG optical activity, whereas collagen I exhibits large chiroptical effects involving both one-electron and excitonic coupl...
Article
We report two novel applications of multiphoton microscopy for pharmacological studies of unstained cardiovascular tissue. First, we show that second harmonic generation (SHG) microscopy of unstained cardiac myocytes can be used to determine the sarcomere length with sub-resolution accuracy, owing to the remarkable contrast of the SHG signal origin...

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