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      Real-TimeReflectance Confocal Microscopy, a Noninvasive Tool for in vivo Quantitative Evaluation of Comedolysis in the Rhino Mouse Model

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          Abstract

          Background: Near-infrared reflectance confocal microscopy (RCM) is a noninvasive tool that provides real-time images of thin virtual horizontal tissue sections. Aims/Methods: We have used a rhino mouse model in combination with topical application of all-trans-retinoic acid and all-trans-retinol to investigate the usefulness of RCM as a noninvasive imaging tool to evaluate comedolysis in vivo and over time. Optical images were correlated with routine histology. Results: Our results demonstrate that RCM in vivo can visualize the process of transformation of utriculi (pseudocomedones) towards a normal-appearing follicular structure during retinoid treatment. The retinoic acid intervention group showed a dose-related response, while the vehicle-treated group did not show utricular changes. Conclusions: RCM represents a useful tool for in vivo morphological and quantitative evaluation of skin utriculi over time and could be used as an adjunct tool to histopathological techniques for comedolysis studies.

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          In vivo confocal scanning laser microscopy of human skin II: advances in instrumentation and comparison with histology.

          M Rajadhyaksha, S. González, J M Zavislan … (1999)
          In 1995, we reported the construction of a video-rate scanning laser confocal microscope for imaging human skin in vivo. Since then, we have improved the resolution, contrast, depth of imaging, and field of view. Confocal images of human skin are shown with experimentally measured lateral resolution 0.5-1.0 microm and axial resolution (section thickness) 3-5 microm at near-infrared wavelengths of 830 nm and 1064 nm; this resolution compares well to that of histology which is based on typically 5 microm thin sections. Imaging is possible to maximum depth of 350 microm over field of view of 160-800 microm. A mechanical skin-contact device was developed to laterally stabilize the imaging site to within +/- 25 microm in the presence of subject motion. Based on these results, we built a small, portable, and robust confocal microscope that is capable of imaging normal and abnormal skin morphology and dynamic processes in vivo, in both laboratory and clinical settings. We report advances in confocal microscope instrumentation and methods, an optimum range of parameters, improved images of normal human skin, and comparison of confocal images with histology.
          Article 0 comments Cited 133 times – based on 0 reviews     Review now
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            In Vivo Confocal Scanning Laser Microscopy of Human Skin: Melanin Provides Strong Contrast

            Milind Rajadhyaksha, Melanie Grossman, Dina Esterowitz … (1995)
            Confocal scanning laser microscopy of live human skin was performed to investigate the correlation of in vivo cellular and morphologic features to histology, the effect of wavelength on imaging, and the role of melanin as a contrast agent. We built a video-rate confocal scanning laser microscope for in vivo imaging of human skin. Using a 100 x microscope objective, we imaged high-contrast optical "sections" of normal skin, vitiliginous skin, and a compound nevus. In vivo "confocal histology" correlated well with conventional histology. The maximum imaging depth increased with wavelength: the epidermis was imaged with visible 400-700-nm wavelengths; the superficial papillary dermis and blood cells (erythrocytes and leukocytes) in the deeper capillaries were imaged with the near infrared 800-900-nm wavelengths. For confocal reflectance imaging, melanin provided strong contrast by increased backscattering of light such that the cytoplasm in heavily pigmented cells imaged brightly. In vivo confocal microscopy potentially offers dermatologists a diagnostic tool that is instant and entirely non-invasive compared to conventional histopathology.
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              Topographic variations in normal skin, as viewed by in vivo reflectance confocal microscopy.

              F Rius, Anjali M. Rajadhyaksha, S. Gonzalez-Sevilla … (2001)
              Near-infrared confocal microscopy is a new tool that provides skin images in vivo, with high resolution and contrast at a specific depth. Regional variations in live human skin viewed by confocal microscope have not been studied so far. In vivo reflectance confocal microscopy was performed in 10 adults (eight males, two females) of various skin phototypes. Six topographic sites were studied in each subject: forehead, cheek, inner and outer forearm surfaces, lower back and leg. Epidermal thickness at suprapapillary epidermal plates and rete pegs was measured during real-time imaging and the number and diameter of epidermal keratinocytes in each epidermal cell layer as well as the characteristics of dermal papillae were defined from the grabbed images. Stratum corneum appeared brighter in sun-exposed than in sun-protected areas and particularly pronounced in heavily pigmented individuals. The epidermal thickness at rete pegs, but not the suprapapillary epidermal plate, was greater in sun-exposed areas than in sun-protected sites except forearm flexor surface. The en face numerical density of granular keratinocytes is greater on the face as compared with all other sites, whereas the surface density of spinous keratinocytes is greater on sun-protected sites. Additionally, the number of basal keratinocytes per millimeter length of dermoepidermal junction is greater in sun exposed areas. Interestingly, the dermal papillae shape varies and their sizes increase in circumference from sun-exposed to sun-protected sites, as observed at a specific depth below the stratum corneum. In summary, our results demonstrate that near infra-red reflectance confocal microscopy is a feasible tool for microscopic analysis of skin morphometry in vivo.
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                Author and article information

                Journal
                Journal ID (publisher-id): SPP
                Journal ID (nlm-ta): Skin Pharmacol Physiol
                Journal ID (doi): 10.1159/issn.1660-5527
                Title: Skin Pharmacology and Physiology
                Publisher: S. Karger AG
                ISSN (Print): 1660-5527
                ISSN (Electronic): 1660-5535
                Publication date Subscription-year: 2007
                Publication date (Print): December 2006
                Publication date (Electronic): 11 October 2006
                Volume: 20
                Issue: 1
                Pages: 29-36
                Affiliations
                aWellman Center, Department of Dermatology, Massachusetts General Hospital, Harvard Medical School, Boston, Mass., and bDermatology Service, Memorial Sloan Kettering Cancer Center, New York, N.Y., USA; cDepartment of Dermatology, Osaka Medical College, Osaka, Japan
                Article
                Publisher ID: 96169 Other ID: Skin Pharmacol Appl Skin Physiol 2007;20:29–36
                DOI: 10.1159/000096169
                PubMed ID: 17035719
                SO-VID: bcd1436b-acae-4dbd-b9f2-82c408f3d2b0
                Copyright © © 2007 S. Karger AG, Basel
                License:

                Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

                History
                Date received : 16 December 2005
                Date accepted : 21 April 2006
                Page count
                Figures: 4, Tables: 1, References: 28, Pages: 8
                Categories
                Subject: Original Paper

                ScienceOpen disciplines: Oncology & Radiotherapy,Pathology,Surgery,Dermatology,Pharmacology & Pharmaceutical medicine
                Keywords: All-trans-retinol,All-trans-retinoic acid,Reflectance confocal microscopy,Rhino mouse, comedones
                Data availability:
                ScienceOpen disciplines: Oncology & Radiotherapy, Pathology, Surgery, Dermatology, Pharmacology & Pharmaceutical medicine
                Keywords: All-trans-retinol, All-trans-retinoic acid, Reflectance confocal microscopy, Rhino mouse, comedones

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