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
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
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: [email protected]
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 protected]
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:
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
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
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The stereo-paired images analyses provide 3 dimensional
perception, which has been used by many previous investiga-
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MITCHEL PAUL GOLDMAN 9339 Genesee Avenue, Suite 300 ; San Diego, California 92121 (Phone: 858-657-1002) (Fax: 858-657-9165) E-Mail: [email protected] BORN: APRIL 5, 1955 (MIAMI BEACH, FLORIDA) EDUCATION Sept 1973 - Jan 1977 Boston University College of Liberal Arts - Biology B.A. 1977 Sept 1978 - June 1982 Stanford University
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