JP3709382B2 - How to differentiate corneocytes - Google Patents
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【０００１】
【発明の属する技術分野】
本発明は、角質細胞の映像及び角質細胞の形状の鑑別法に関する。
【０００２】
【従来の技術】
化粧料を使用するにあたり、重要な事項は適切な化粧料を選択することである。この様な適切な化粧料の選択を行う必要条件としては、皮膚の状態或いは特性を正しく鑑別することが挙げられる。この様な皮膚の状態或いは特性を正しく行うための技術としては、顔の頬などの部位より粘着テープなどを用いて、ストリッピングにより角質細胞を採取し、ゲンチアナバイオレット等の染色剤を用いて、角質細胞を染色し、角質細胞の境界を明確にし、角質細胞の面積或いは体積を測定して、その値を指標にする技術が既に確立されている。即ち、面積或いは体積が小さいほど、角質細胞が充分成熟しない内に最外部に上がってきてしまっており、皮膚バリア機能は低く、肌の状態が悪いと鑑別され、面積が大きいほど皮膚バリア機能が高く、肌の状態が良いと鑑別される技術である。これに加えて、どの様な形で角質細胞が剥離されてきているか、言い換えれば、角質細胞がまとまっていくつか取れている重層剥離が頻繁に観察されるのか、殆どの角質細胞が１個１個離れて剥離しているのかによって、過敏な肌なのか否かが鑑別でき、これらをあわせて考察することにより、肌特性が的確に鑑別できるわけである。この様な鑑別の基礎となるのは、何れも角質細胞の形状であり、かかる形状を正確に知るには、これまでは先に述べたように染色剤を用いて染色する工程を経なければならなかった。この工程には数日の時間を要し、適時的な観察を販売現場で行うことは極めて困難の伴うものであった。加えて、染色に於ける染色剤を含む廃液の処理は環境問題を考慮する上で、大きな課題の一つであった。言い換えれば、染色工程無しに角質細胞の形状を的確に把握できる手段の開発が望まれていた。
【０００３】
一方、皮膚より採取した角質細胞の形状の鑑別法であって、当該角質細胞を紫外線下観察される像を、画像として取り込むことを特徴とする、角質細胞の形状の鑑別法は全く知られていないし、この様な手段により、染色工程無しに角質細胞の形状を的確に把握できることも全く知られていない。従って、この様な手段を取ることにより、適時的に迅速な肌の鑑別が、環境にも優しく行えることも全く知られていない。
【０００４】
【発明が解決しようとする課題】
本発明は、この様な状況下為されたものであり、染色工程無しに角質細胞の形状を的確に把握できる手段を提供することを課題とする。
【０００５】
【課題の解決手段】
本発明者らは、この様な状況に鑑みて、染色工程無しに角質細胞の形状を的確に把握できる手段を求めて、鋭意研究努力を重ねた結果、皮膚より採取した角質細胞を紫外線下、顕微鏡乃至はビデオマイクロスコープを介して撮影した映像が、角質細胞の形状把握に好適であることを見出し、発明を完成させるに至った。即ち、本発明は以下に示す技術に関するものである。
（１）皮膚より採取した角質細胞の形状の鑑別法であって、当該角質細胞を紫外線下観察される像を、画像として取り込むことを特徴とする、角質細胞の形状の鑑別法。
（２）使用する紫外線が、３４０〜３８０ｎｍに極大波長を持つものであることを特徴とする、（１）に記載の角質細胞の形状の鑑別法。
（３）画像取り込みが、顕微鏡乃至はビデオマイクロスコープによるものであることを特徴とする、（１）又は（２）に記載の角質細胞の形状の鑑別法。
（４）角質細胞が、顔より粘着面を有する媒体によって採取されたものであることを特徴とする、（１）〜（３）何れか１項に記載の角質細胞の形状の鑑別法。
（５）粘着面を有する媒体が、粘着組成物を塗工したプラスチック製のディスクであることを特徴とする、（４）に記載の角質細胞の形状の鑑別法。
（６）皮膚の状態の鑑別を目的とすることを特徴とする、（１）〜（５）何れか１項に記載の角質細胞の形状の鑑別法。
（７）皮膚より採取した角質細胞の形状の鑑別の為の映像であって、当該角質細胞を紫外線下、顕微鏡乃至はビデオマイクロスコープを介して撮影した映像。
（８）使用する紫外線が、３４０〜３８０ｎｍに極大波長を持つものであることを特徴とする、（７）に記載の映像。
（９）ＬＥＤ光源を紫外線の光源とすることを特徴とする、（７）又は（８）に記載の映像。
（１０）皮膚の状態の鑑別を目的とすることを特徴とする、（７）〜（９）何れか１項に記載の映像。
（１１）（７）〜（１０）何れか１項に記載の映像を用いて、（１）〜（６）何れか１項に記載の角質細胞の鑑別法により、角質細胞の形状を鑑別し、これを指標とすることを特徴とする、皮膚特性の鑑別法。
以下に本発明について更に詳細に説明を加える。
【０００６】
【発明の実施の形態】
（１）本発明の角質細胞の形状の鑑別の為の映像
本発明の角質細胞の形状の鑑別の為の映像は、皮膚より採取した角質細胞の形状の鑑別の為の映像であって、当該角質細胞を紫外線下、顕微鏡乃至はビデオマイクロスコープを介して撮影したことを特徴とする。本発明の映像作成のための角質細胞標本としては、これまで知られている、染色を必須とする角質細胞標本の作り方に準拠し、染色工程を省いた形で作成することができる。即ち、顔の頬などの部位より粘着テープや粘着剤を塗工したディスクなどの粘着支持体を用いて、ストリッピングにより角質細胞を採取したものが好適に例示できる。粘着支持体としては、均一性の点でディスクに粘着剤を塗工した形態が更に好ましく例示できる。この際、更にもう一度粘着支持体を用いて角質細胞を転写し、角質細胞の表面側の面を上面にし、この面を観察することもできる。この様な転写を行うことは、角質細胞を均一な一層にすることができるので、角質細胞の形状の理解にとっては好ましい。かくして得られた角質標本に紫外線を照射し、その結果、紫外線の反射、透過或いは蛍光によって生じる映像が本発明の映像である。即ち、本発明の映像としては、反射映像でも、透過映像でも或いは蛍光映像でも構わない。反射映像であれば、標本に対してムラの無いように極近傍の周囲から一様に紫外線を照射し、中心より映像を取得すればよい。透過画像であれば、予め粘着支持体自身を使用する紫外線透過性のもので調整し、標本の裏側より紫外線を照射し、透過光が結んだ映像を取得すればよい。蛍光画像を得るには、上部乃至は下部より紫外線を照射し、反射乃至は透過の紫外線を必要に応じてフィルターによって除去し、蛍光によって生じる可視光のみを集めて映像を取得すればよい。この様な映像を作るために照射する紫外線としては長波長でも短波長でも構わないが、特に好ましいものは３４０〜３８０ｎｍに極大波長を持つものである。この様な紫外線を提供する光源としてはＬＥＤ光源（ライトエミッティング・ダイオード光源）が好ましく例示できる。この様な映像取得に特に好ましい機器としては蛍光顕微鏡が挙げられる。蛍光顕微鏡においては、下部より標本に紫外線を照射し、透過光乃至は蛍光をレンズで集め、映像を取得することができる。かかる映像は、デジタルカメラなどを介してデジタル画像として取り込むことができる。又、ビデオマイクロスコープのビデオカメラの部分を改良して、カメラの周りに紫外線光源を配し、遮光性のアタッチメントで光源とカメラを覆い、光源からの光の反射光のみを拾うような形状にして前記の角質細胞標本の映像を得ることもできる。この様にして得られる本発明の映像においては角質細胞は明確にその境界が映し出されるため、染色無しに、本発明の映像によって、その形状を把握することができる。かかる映像は、その境界も鮮明であるため、コンピューターソフトを利用して、面積などを自動的に計測することもできる。
【０００７】
（２）本発明の角質細胞の鑑別法、皮膚特性の鑑別法
本発明の角質細胞の鑑別法は、前記本発明の角質細胞の形状の鑑別の為の映像を用いることを特徴とする。角質細胞の形状により、皮膚特性、例えば、肌の刺激に対する感受性、バリア機能を含む肌の状態、肌の老化の程度を表す肌年齢などを知ることができる。この様な情報を得るための角質細胞の形状としては、角質細胞の配列の規則性、大きさ（体積或いはその便宜的代替値としての面積）、一層で剥離してくるか、数層が重なったまま剥離してくるかという、角質細胞の剥離のパターン等が例示できる。本発明の映像を用いることにより、これらの何れもが、角質細胞標本を染色して得た画像と同程度の精度で鑑別、測定することができる。配列規則性であれば、個々の細胞の形がどの程度六角形になっているかを肉眼で判定したりすることにより鑑別できるし、大きさであれば、二値化などの処理を施した後に、画像処理ソフトを使用することにより自動計測することができる。剥離パターンは、配列規則性同様に、肉眼判定できるし、或いは、画像の濃さ（或いは明るさ）などを、輝度から算出し、推定することもできる。この様な方法については、染色標本において使用されている技術をそのまま応用することができる。又、この様に鑑別された角質細胞の形状をもとに、従来知られている角質細胞と皮膚特性の関係を利用して皮膚特性を鑑別することもできる。かかる皮膚特性の鑑別が、本発明の皮膚特性の鑑別法である。この様な技術で、既に知られているものとしては、特開２００２−１７６８８号、特開２００１−１０８６７４号、特開２００１−１３１３８号、特開２０００−１１６６２３号、特開平１１−３０４７９８号、特開平１１−２９９７９２号、特開平７−２０９２９２号等が例示でき、これらの何れもが使用することができる。従来の染色標本法が標本調整に、染色剤などを用いることができる環境において数時間要していたのに比べ、本発明の鑑別法は、通常の環境で、ほぼリアルタイムに鑑別を行うことができる。即ち、染色剤などを用いることができる環境への角質細胞の移送時間を加味すれば、従来法に比して、本発明の鑑別法では、数日間の時間短縮が行える。加えて、染色剤を必要としないことから廃水処理の必要性もなく、環境保全の観点からも本発明の鑑別法は好ましい。この様に、皮膚特性を鑑別することにより、適切な化粧料の選択が可能となり、誤った化粧料の選択による、トラブルの発生や無駄な投資を避けることも可能である。又、本発明の鑑別法は、採取した角質細胞には非侵襲的であるので、角質細胞の形状の鑑別を行った後に、種々の染色標本に供することも可能であり、従来の方法に比して、更に多面的な情報の入手も可能となる。
【０００８】
【実施例】
以下に、実施例を挙げて、本発明について更に詳細に説明を加えるが、本発明がかかる実施例にのみ限定を受けないことは言うまでもない。
【０００９】
＜実施例１＞
粘着テープを頬部に貼付し、上から均一に抑えた後、剥離し、これを可視光の透過条件（通常の光学顕微鏡）及び紫外線の透過条件（中心波長３６０ｎｍ；蛍光顕微鏡）で観察を行った。これを画像として取り込んだものを図１（図面代用写真）に示す。可視光透過条件においては、角質細胞の境界は極めて不明瞭であり、その形状を認知することはできないが、紫外線透過条件においては、明瞭に角質細胞の形状が認知でき、面積などが測定できることがわかる。
【００１０】
＜実施例２＞
実施例１の紫外線透過条件での映像より角質細胞の平均面積を求めた。又、この角質細胞標本をゲンチアナバイオレットで染色し、従来法に従って同様に角質細胞の平均面積を求めた。結果を表１にしめす。これより、本発明の映像より、従来法と同程度の精度で面積が測定できることがわかる。
【００１１】
【表１】
【発明の効果】
本発明によれば、染色工程無しに角質細胞の形状を的確に把握できる手段を提供することができる。
【図面の簡単な説明】
【図１】 実施例１の可視光透過映像と本発明の紫外線透過映像の比較を示す図である。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an image of corneocytes and a method for distinguishing the shape of corneocytes.
[0002]
[Prior art]
In using cosmetics, an important matter is to select appropriate cosmetics. A necessary condition for selecting such an appropriate cosmetic is to correctly distinguish the state or characteristics of the skin. As a technique for correctly performing such skin condition or characteristics, using an adhesive tape or the like from the face cheeks, collecting keratinocytes by stripping, using a stain such as gentian violet, A technique has already been established for staining corneocytes, clarifying the boundaries of the corneocytes, measuring the area or volume of the corneocytes, and using the value as an index. In other words, the smaller the area or volume, the keratinocytes have risen to the outermost part before they are sufficiently matured, the skin barrier function is low, and it is identified that the skin condition is bad. It is a technology that is distinguished by high skin condition. In addition to this, how keratinocytes have been exfoliated, in other words, whether stratified exfoliation in which several keratinocytes are taken together is frequently observed, most keratinocytes are one by one. Whether the skin is sensitive or not can be discriminated depending on whether it is peeled apart, and the skin characteristics can be accurately discriminated by considering them together. The basis of such differentiation is the shape of the keratinocytes, and in order to know the shape accurately, it has been necessary to go through the process of staining with a stain as described above. did not become. This process took several days, and it was extremely difficult to conduct timely observations at the sales site. In addition, the treatment of waste liquid containing dyeing agent in dyeing has been one of the major issues in consideration of environmental problems. In other words, it has been desired to develop a means capable of accurately grasping the shape of keratinocytes without a staining step.
[0003]
On the other hand, there is no known method for distinguishing the shape of keratinocytes, which is a method for distinguishing the shape of keratinocytes collected from the skin, characterized in that an image of the keratinocytes observed under ultraviolet rays is captured as an image. In addition, it is not known at all that the shape of corneocytes can be accurately grasped by such means without a staining step. Therefore, it is not known at all that taking such a measure enables timely and rapid skin discrimination to be performed in an environmentally friendly manner.
[0004]
[Problems to be solved by the invention]
The present invention has been made under such circumstances, and an object of the present invention is to provide means for accurately grasping the shape of the corneocytes without a staining step.
[0005]
[Means for solving problems]
In light of such circumstances, the present inventors have sought for means capable of accurately grasping the shape of keratinocytes without a staining step, and as a result of intensive research efforts, the keratinocytes collected from the skin under ultraviolet light, The inventors have found that an image photographed through a microscope or a video microscope is suitable for grasping the shape of a keratinocyte, and have completed the invention. That is, the present invention relates to the following technique.
(1) A method for distinguishing the shape of a keratinocyte collected from the skin, which comprises capturing an image obtained by observing the corneocyte under ultraviolet light as an image.
(2) The method for distinguishing the shape of a keratinocyte according to (1), wherein the ultraviolet ray to be used has a maximum wavelength at 340 to 380 nm.
(3) The method for distinguishing the shape of a keratinocyte according to (1) or (2), wherein the image capturing is performed by a microscope or a video microscope.
(4) The method for distinguishing the shape of a keratinocyte according to any one of (1) to (3), wherein the keratinocyte is collected by a medium having an adhesive surface from the face.
(5) The method for distinguishing the shape of keratinocytes according to (4), wherein the medium having an adhesive surface is a plastic disc coated with an adhesive composition.
(6) The method for distinguishing the shape of a keratinocyte according to any one of (1) to (5), characterized in that the purpose is to distinguish a skin state.
(7) An image for distinguishing the shape of keratinocytes collected from the skin, which is taken through a microscope or a video microscope under ultraviolet light.
(8) The image according to (7), wherein the ultraviolet ray used has a maximum wavelength at 340 to 380 nm.
(9) The image according to (7) or (8), wherein the LED light source is an ultraviolet light source.
(10) The image according to any one of (7) to (9), characterized in that the purpose is to distinguish a skin state.
(11) Using the image described in any one of (7) to (10), the shape of the keratinocytes is identified by the differentiation method for keratinocytes described in any one of (1) to (6). A method for distinguishing skin characteristics, characterized by using this as an index.
The present invention will be described in further detail below.
[0006]
DETAILED DESCRIPTION OF THE INVENTION
(1) Image for distinguishing the shape of keratinocytes of the present invention The image for distinguishing the shape of keratinocytes of the present invention is an image for distinguishing the shape of keratinocytes collected from the skin. The keratinocytes are photographed under a UV ray through a microscope or a video microscope. The keratinocyte specimen for creating an image of the present invention can be prepared in a form that omits the staining step in accordance with the known method of making a keratinocyte specimen that requires staining. That is, a sample obtained by stripping keratinocytes by stripping using an adhesive support such as an adhesive tape or a disk coated with an adhesive from a part such as the cheek of the face can be preferably exemplified. As the adhesive support, a form in which an adhesive is applied to the disk in terms of uniformity can be further exemplified. At this time, the keratinocytes can be transferred once again using the adhesive support, and the surface on the surface side of the keratinocytes can be used as the upper surface, and this surface can be observed. Performing such transcription is preferable for understanding the shape of the keratinocytes because the keratinocytes can be made uniform in one layer. The image of the present invention is an image generated by irradiating ultraviolet rays to the horny specimen thus obtained and, as a result, reflection, transmission or fluorescence of the ultraviolet rays. That is, the image of the present invention may be a reflected image, a transmitted image, or a fluorescent image. In the case of a reflected image, the sample may be acquired from the center by uniformly irradiating ultraviolet rays from the vicinity of the vicinity so that the specimen is not uneven. In the case of a transmission image, it is only necessary to adjust in advance with an ultraviolet ray transmissive material using the adhesive support itself, and to irradiate ultraviolet rays from the back side of the specimen to obtain an image in which the transmitted light is tied. In order to obtain a fluorescent image, it is only necessary to irradiate ultraviolet rays from the upper part or the lower part, remove the reflected or transmitted ultraviolet rays with a filter as necessary, and collect only visible light generated by fluorescence to obtain an image. The ultraviolet rays irradiated for producing such an image may be long wavelength or short wavelength, but particularly preferable are those having a maximum wavelength at 340 to 380 nm. As a light source for providing such ultraviolet rays, an LED light source (light emitting diode light source) can be preferably exemplified. A particularly preferable apparatus for such image acquisition is a fluorescence microscope. In a fluorescence microscope, an image can be acquired by irradiating a specimen with ultraviolet rays from the lower part and collecting transmitted light or fluorescence with a lens. Such video can be captured as a digital image via a digital camera or the like. In addition, the video camera part of the video microscope has been improved so that an ultraviolet light source is placed around the camera, the light source and the camera are covered with a light-shielding attachment, and only the reflected light from the light source is picked up. Thus, an image of the keratinocyte specimen can be obtained. In the image of the present invention obtained in this way, the boundary of the keratinocytes is clearly projected, so that the shape can be grasped by the image of the present invention without staining. Since the boundary of such an image is clear, the area and the like can be automatically measured using computer software.
[0007]
(2) Corneocyte differentiation method and skin characteristic discrimination method of the present invention The corneocyte differentiation method of the present invention is characterized by using an image for differentiation of the shape of the keratinocytes of the present invention. From the shape of the keratinocytes, it is possible to know skin characteristics such as sensitivity to skin irritation, skin condition including a barrier function, skin age indicating the degree of skin aging, and the like. The shape of the keratinocytes to obtain such information is the regularity and size of the keratinocyte array, the size (volume or area as a convenient alternative value), one layer peeling or several layers overlapping The pattern of exfoliation of keratinocytes, such as whether or not to exfoliate as is, can be exemplified. By using the image of the present invention, any of these can be distinguished and measured with the same degree of accuracy as an image obtained by staining a keratinocyte specimen. If the arrangement is regular, it can be distinguished by judging with the naked eye how much the shape of each cell is hexagonal, and if it is large, after performing processing such as binarization Automatic measurement can be performed by using image processing software. As with the arrangement regularity, the peeling pattern can be determined with the naked eye, or the darkness (or brightness) of the image can be calculated and estimated from the luminance. For such a method, the technique used in the stained specimen can be applied as it is. Further, based on the shape of the corneocytes thus differentiated, it is also possible to distinguish the skin characteristics by utilizing the conventionally known relationship between the corneocytes and the skin characteristics. Such discrimination of skin characteristics is the skin characteristic discrimination method of the present invention. As such techniques, those already known are disclosed in JP 2002-17688, JP 2001-108674, JP 2001-13138, JP 2000-116623, JP 11-304798, JP-A-11-299792, JP-A-7-209292 and the like can be exemplified, and any of these can be used. The conventional stained specimen methods specimen adjustment, compared to it takes several hours in an environment which can be used as the staining agent, differentiation method of the present invention, under normal circumstances, it is carried out differential in near real time it can. In other words, if the time for transporting corneocytes to an environment where a staining agent or the like can be used is taken into consideration, the differentiation method of the present invention can shorten the time for several days as compared with the conventional method. In addition, since no dyeing agent is required, there is no need for wastewater treatment, and the discrimination method of the present invention is preferable from the viewpoint of environmental conservation. Thus, by distinguishing the skin characteristics, it is possible to select an appropriate cosmetic, and it is also possible to avoid troubles and wasteful investment due to incorrect selection of the cosmetic. In addition, since the differentiation method of the present invention is non-invasive to the collected keratinocytes, it can be used for various stained specimens after differentiation of the shape of the keratinocytes, compared with the conventional method. In addition, it is possible to obtain more multifaceted information.
[0008]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples, but it goes without saying that the present invention is not limited to such examples.
[0009]
<Example 1>
Adhesive tape is affixed to the cheek, and after being suppressed uniformly from above, it is peeled off and observed under visible light transmission conditions (normal optical microscope) and ultraviolet light transmission conditions (central wavelength 360 nm; fluorescence microscope). It was. The image captured as an image is shown in FIG. 1 (drawing substitute photo). Under visible light transmission conditions, the stratum corneum boundary is very unclear and its shape cannot be recognized, but under ultraviolet light transmission conditions, the shape of the stratum corneum can be clearly recognized and the area can be measured. Understand.
[0010]
<Example 2>
The average area of the keratinocytes was determined from the image of Example 1 under the ultraviolet transmission condition. Further, this keratinocyte specimen was stained with gentian violet, and the average area of the keratinocytes was similarly determined according to the conventional method. The results are shown in Table 1. From this, it can be seen from the image of the present invention that the area can be measured with the same accuracy as the conventional method.
[0011]
[Table 1]
【The invention's effect】
ADVANTAGE OF THE INVENTION According to this invention, the means which can grasp | ascertain the shape of a keratinocyte exactly without a dyeing process can be provided.
[Brief description of the drawings]
FIG. 1 is a diagram showing a comparison between a visible light transmission image of Example 1 and an ultraviolet transmission image of the present invention.
Claims (11)
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| JP2002120616A JP3709382B2 (en) | 2002-04-23 | 2002-04-23 | How to differentiate corneocytes |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101622538B (en) * | 2007-02-14 | 2012-09-26 | 宝丽化学工业有限公司 | Method of supporting the differentiation of horny layer cells |
| CN102819732A (en) * | 2007-02-14 | 2012-12-12 | 宝丽化学工业有限公司 | Method and computer for identifying keratinocytes |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US9750449B2 (en) * | 2003-12-12 | 2017-09-05 | Johnson & Johnson Consumer Inc. | Method of assessing skin |
| JP2006017688A (en) * | 2004-06-01 | 2006-01-19 | Pola Chem Ind Inc | Differentiation method for subcorneal cell by autofluorescence |
| JP4549108B2 (en) * | 2004-06-11 | 2010-09-22 | ポーラ化成工業株式会社 | How to measure skin flexibility |
| JP4818704B2 (en) * | 2005-12-15 | 2011-11-16 | ポーラ化成工業株式会社 | Image color adjustment method |
| JP5065237B2 (en) * | 2008-12-26 | 2012-10-31 | 常盤薬品工業株式会社 | Noninvasive evaluation method of local pathology of atopic dermatitis |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101622538B (en) * | 2007-02-14 | 2012-09-26 | 宝丽化学工业有限公司 | Method of supporting the differentiation of horny layer cells |
| CN102819732A (en) * | 2007-02-14 | 2012-12-12 | 宝丽化学工业有限公司 | Method and computer for identifying keratinocytes |
| CN102819732B (en) * | 2007-02-14 | 2016-02-10 | 宝丽化学工业有限公司 | Support method and the computing machine of resolution angle cell plastid |
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