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12-scanning(jh)한국현미경학회지 제40권 제4호 2010 Korean J. Microscopy 40(4), 267 270(2010) Scanning Electron Microscopic Observation of
Human Skin Replica
Yeon Seung Rhyu, Ye Ji Chung*, Chang Sub Uhm1,* composed of five layers called stratum corneum, stratum luci- Buram High School, Seoul 139-861, Korea dum, stratum glanulosum, stratum spinosum, and stratum ba- 1Department of Anatomy, College of Medicine, sale. Its other functions are insulation, temperature regulation, Seoul 136-705, Korea University, Korea sensation, prevention of excessive water loss, and the produc- (Received October 22, 2010; Revised December 6, 2010; Accepted tion of vitamin D. The role of skin as aesthetics and communi- December 7, 2010) cation is also stressed in modern society. Under those layers ABSTRACT :
there are dermis and hypodermis (Junqueira & Carneiro, 2005).
The skin is the largest organ of the integument sys- tem whose surface is closely related with many physiological and The surface of the skin, or epidermal surface, shows a net- pathological conditions. Various methods are used to understand like structure. This structure is the irregular and roughly shaped the structural and functional status of human skin. We would like polygons, most often resembling a square. The edges of the to present usefulness of scanning electron microscopic (SEM) polygons define location of furrows or micro lines (Mark, 1983).
observation of skin replica and its significance of training module As the skin texture becomes rough, the wrinkles tend to deepen for a novice.
and the surface mesh-like structure changes its arrangement or The silicon replicas from several regions of the body (hand, finger, disappears (Gilchrest, 1996).
forearm, lip, and face) were casted by applying Exafine® mixture.
The positive replicas were prepared by applying EPON 812 mix- Some noninvasive methods such as magnifier, Wood's lamp, ture on negative silicon replicas. Some of the negative silicon repli- and skin scope have been used for routine evaluation skin sta- cas were cut with a razor blade and surface profiles were observed.
tus in vivo. Although a skin biopsy provides detailed informa- The negative and positive replicas were coated with platinum and tion on the skin, it is an invasive method that needs serious were observed under the scanning electron microscope.
consideration of a dermatologist. Various topographic analyses We could investigate the detailed structures of the human skin have been performed to understand the status of skin surface surface without any physical damage to the subject. The positive replicas depicted real surface structure of the human skin vividly.
(Fischer et al., 1999; Lagadre et al., 2005). The negative or The cross sectional view of the negative silicon replicas provided positive replicas of skin surface were analyzed by profilometry surface profile clearly. (mechanical, optical, laser, transmission, or interference). Objec- The scanning electron microscopic observation of the human skin tive assessment with profilometry was applied to various phy- replicas would be useful to study skin surface structures and to siological, pathological and clinical studies (Choi and Oh, 1997; evaluate medical and esthetical applications. (유연승, 정예지, 엄 Son et al., 2005; Dessy et al., 2007) 창섭: 사람피부 모사판의 주사전자현미경적 관찰) In this study, we present the usefulness of scanning electron Keywords : Education, Profilometry, Replica, Skin microrelief,
microscopic (SEM) observation of the skin replica and its sig- Stereo pair image nificance as a training module for a novice.
Four people voluntarily provided replicas. The purpose and procedures of the study were fully explained to the participants.
The skin is a covering of the body. It is the largest organ of The skin surface for replica was swapped with alcohol sponge.
the human body that takes up to 16 percent of the weight and After completely dried, the mixture of silicone mixture (Exa- acts as a primary defense against external hostiles and excessive fine®, GC Co, Japan) was spread over the region of interest water loss. The outer layer of skin is called epidermis which is and waited for 3 5 minutes to solidify, and the cast was re- * Correspondence should be addressed to Ye Ji Chung, Buram High School, Junggye Bon Dong, Nowon Gu, Seoul 139-861, Korea. Ph.: (02) 935-7300, Fax: (02)936-9100, E-mail: firstname.lastname@example.org or Prof. Chang Sub Uhm, Department of Anatomy, College of Medicine, Korea University, 126-1 Anam Dong, SeongbukGu, Seoul 136-705, Korea. Ph.: (02) 920-6150, Fax: (02) 929-5696, E-mail: email@example.com
Korean J. Microscopy Vol. 40, No. 4, 2010 moved carefully from the skin surface. The replicas were col- sional structure of the skin more vividly, three different tilting lected from forearm, back of hand, finger pad, dorsal surface ) were used for observation. Two stereo- of finger, outside of eye and upper lip. paired images were selected and anaglyphic stereo-image was The negative silicon replicas were used as molds to produce constructed by Anamaker® program (Takashi Sekitani, http:// positive replicas. The Epon mixture (Epon 812: 48.5 mL, DDSA: www.stereoeye.jp). 18.5 mL, NMA: 33.0 mL, and DMP-30: 1.5 mL) was poured The negative silicon replica of the dorsum of hand has a onto the negative replicas and put into an oven of 60� mirror image of the original skin surface. It revealed that the days. Some of the negative silicon replicas were cut with a skin surface was composed of large polygons containing small razor blade vertically across the horizontal furrows of the hand polygons inside (Fig. 1A). The positive Epon replica showed a and finger. The cut specimens were attached on a stub with real surface structure of the hand, which is at the same orien- carbon tape, and platinum-coated with an ion coater (IB-5 ion tation of the skin surface (Fig. 1B). The sectional plane along coater, Eiko Co., Japan). The replicas were investigated under the major horizontal skin furrow of the hand silicon replica SEM (S-4700S, Hitachi, Japan). To understand the three dimen- revealed profiles of the furrows reminiscent of a mountain Fig. 1. Scanning electron microscopic image of silicon replica (A) prepared from dorsum of hand, which is a negative cast of superficial skin, the
positive replica (B) can be produced by applying epoxy resin on the negative replica. The cross sectional view of a negative silicon replica reveals
detailed profiles of the finger surface (C, D). The positive replica of finger print shows typical ridge and furrow of the print (E), the sweat pores
(arrow) are observed on the ridge of the finger print (F: inset of E).
Rhyu YS et al. : SEM Observation of Human Skin Replica Fig. 2. Stereo pair images (A1: 0�tilted, A2: 5�tilted) of positive replica of upper lip show three dimensional configuration of hair and their
underlying skin. Anaglyphic image was constructed with a pair of 5�
-tilted micrographs and stereoscopic features of hair are observed under red- blue glasses (A3). Anaglyphic images of forearm (B), dorsal surface of finger (C) and outside of eye (D) show different microrelief according tothe anatomical location.
chain. The top of a mountain in the replica falls on is the bot- showed usefulness of comparative observation of the negative tom line of a furrow and overall shape of the mountain reflects and positive replicas providing complementary image of the the dimension of the skin furrow (Fig. 1C, D). The positive skin surfaces. The positive replica provides imitation of the replica obtained at the pad of a fingertip showed fingerprint real skin surface very closely without any physical damage to pattern clearly, with smooth ridges and shallow grooves (Fig.
1E). At the higher magnification, the openings of sweat pores Especially, the method observing the vertically cut silicon were clearly demonstrated on the ridge (Fig. 1F).
replicas would be highly useful for the analysis of the surface The positive replica image from upper lip shows hair shafts of the skin furrows. The cross sectional view of the silicon and their ground surface structure (Fig. 2A). Stereo pair images replica turned out to be a unique approach demonstrating an revealed three dimensional configuration of the skin surface exact profile of the skin surface under the SEM. Thanks to the (Fig. 2A1, 2). Anaglyphic images constructed from stereo-pair high resolution image of the skin profile, we could analyze of different tilting angle and stereoscopic features of the skin exact dimension of the skin furrow, which would be important can be observed under red-blue glasses (Fig. 2A3). Anaglyphic parameter to understand skin surface texture properties.
images from forearm, dorsum of hand and outside the eye There are some advantages of using a replica, although it show characteristic surface structures according to their anato- might lose some original details and make artifacts during mical locations. preparations. The replica is an inorganic compound, which The skin silicon replica has been used for the evaluation of does not affected by temperature, moisture, and biochemical many physiological and pathological conditions (Choi et al., changes such as rotting happening in the biological samples.
1997; Son et al., 2005; Zou et al., 2005). The profiles of skin Also the replica can be cut in any direction as needed, can be replica provide an objective assessment of clinical interven- observed at any time after collection without damage to the tions such as Botulinum toxin injection (Grove et al., 1991; subject. And replica does not require fixation step, which is Dessy et al., 2007) or application of anti-wrinkle compounds an essentially required step for biological sample preparation (Rbino et al., 2005; Kawada et al., 2008). In this study, we for SEM, a novice can learn scanning electron microscopy with Korean J. Microscopy Vol. 40, No. 4, 2010 easy. Therefore, SEM study with replica system could be used J Dermatol 135 : 867-875, 1996.
as an education module for beginners to have a concept of Grove GL, Grove MJ, Leyden JJ, Lufrano L, Schwab B, Perry BH, microscopic system. Thorne EG: Skin replica analysis of photodamaged skin after therapy with tretinoin emollient cream. J Am Acad Dermatol 25 : The stereo-paired images analyses provide 3 dimensional 231-237, 1991.
perception, which has been used by many previous investiga- Hortolà P: Using digital anaglyphy to improve the relief effect of tors (Kim et al., 2007; Hortola, 2009). Although experts can SEM micrographs of bloodstains. Micron 40 : 409-412, 2009.
perceive 3 dimensional configuration from stereo-paired images Junqueira LCU, Carneiro J: Basic histology: text & atlas 11th ed, without any additional glasses, the beginner requires stereo- McGraw-Hill, New York, pp. 360-366, 2005.
scopic glasses such as red-blue glasses, polarized glasses.
Kawada A, Konishi N, Oiso N, Kawara S, Date A: Evaluation of Anaglyphic image is one of the useful methods to present 3 anti-wrinkle effects of a novel cosmetic containing niacinamide.
J Dermatology 35 : 637-642, 2008.
dimensional images, because it contains information from stereo Kim JW, Lee SJ, Rhyu IJ: Construction of anaglyphic stereo pair paired images on the single picture appears on the paper or the image using Adobe Photoshop® program. Korean J Electron computer monitor. Microscopy 37 : 143-146, 2007. (Korean) In conclusion, this SEM research of skin replica will provide Lagarde JM, Rouvrais C, Black D: Topography and anisotropy of an objective way to evaluate the human skin surface status the skin surface with ageing. Skin Res Technol 11 : 110-119, related with various skin related applications. Due to the low level of handling procedure, replica system with SEM could Marks R: Mechanical properties of the skin. In: Goldsmith LA, ed.
be used as an education module for students to have a concept Biochemistry and Physiology of the Skin, pp. 1237-1254, Ox- ford University Press, New York, 1983. Park SY, Ha SH, Yu DS, Son SW, Kim DJ, Kim IH, Moon JS, Kim MK, Oh CH: Quantitative evaluation of severity in psoriatic lesions using three-dimensional morphometry. Exp Dermatol 13 : 223-228, 2004.
Rubino C, Farace F, Dessy LA, Sanna MP, Mazzarello V: A pros- Choi HC, Oh CH: Evaluation of skin furrows in the ageing process pective study of anti-aging topical therapies using a quantitative using an image analysis system. Kor J Dermatol 35(2) : 292-302, method of assessment. Plast Reconstr Surg 115 : 1156-1162, Dessy LA, Mazzocchi M, Rubino C, Mazzarello V, Spissu N, Scu- Son SW, Park SY, Ha SH, Park GM, Kim MG, Moon JS, Yoo DS, deri N: An objective assessment of botulinum toxin A effect on Oh CH: Objective evaluation for severity of atopic dermatitis superficial skin texture. Ann Plast Surg 58 : 469-473, 2007.
by morphologic study of skin surface contours. Skin Research Fischer TW, Wigger-Alberti W, Elsner P: Direct and non-direct Technology 11 : 272-280, 2005.
measurement techniques for analysis of skin surface topography.
Zou Y, Song E, Jin R: Age-dependent changes in skin surface asses- Skin Pharmacol Appl Physiol 12 : 1-11, 1999.
sed by a novel two-dimensional image analysis. Skin Res Tech- Gilchrest BA: A review of skin ageing and its medical therapy. Br nol 15 : 399-406, 2009.
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