A novel in vitro stripping method to study geometry of corneocytes with fluorescent microscopy: example of aging skin
A novel in vitro stripping method to study geometry of corneocytes with fluorescent microscopy: example of aging skin
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DOI：
10.1111/j.1600-0846.2009.00369.x
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发表时间：
2009-11-01
影响因子：
2.2
通讯作者：
Sokolov, I.
中科院分区：
文献类型：
--
作者：
Guz, N. V.;Gaikwad, R. M.;Sokolov, I.
背景/目的改进剥片法，使体外培养的人皮肤角质层细胞能够高分辨率荧光显示。为了验证该方法，测量的角质细胞面积的皮肤薄片收集从不同年龄的individuals.Materials和methodsTwo互补的荧光染料依次使用。首先，剥离带的粘合剂层用阳离子染料（若丹明640）染色。这条胶带是用来收集皮肤碎片的。然后用阴离子染料（荧光素）对胶带和收集的薄片进行染色。暴露于第二染色的胶带的荧光由于染料的相互猝灭而显著降低。13名健康的6-86岁男性参与验证该方法。测量是在反手和前臂。ResultsThe方法允许通过荧光显微镜的高分辨率成像的角质细胞。无论是绝对面积和依赖于个人年龄的角质细胞面积是在良好的协议与以前报道的data.ConclusionThe开发的方法是快速和容易。它需要与个人进行最少的互动，并允许使用各种可能不安全但对成像有益的荧光染料。它可以用于人体或动物身体的任何部位。
Background/purposeTo develop modification of stripping method allowing high-resolution fluorescent visualization of corneocytes of human skin in vitro. To validate the method, the measured corneocyte areas on skin flakes are collected from individuals of different ages.Materials and methodsTwo complimentary fluorescent dyes were used sequentially. First the adhesive layer of the stripping tape was stained with a cationic dye (rhodamine 640). This tape was used to collect skin flakes. Then both the tape and collected flakes were stained with anionic dye (fluorescein). The fluorescence of the adhesive tape exposed to the second staining is substantially decreased due to the mutual quenching of the dyes. Thirteen healthy, 6-86-year-old males participated to validate the method. The measurements were done on backhand and forearm.ResultsThe method allows high-resolution imaging of corneocytes by means of fluorescent microscopy. Both absolute areas and the dependence of corneocyte areas on the individual age are in good agreement with the data reported previously.ConclusionThe developed method is fast and easy. It requires minimum interaction with the individual and allows using a broad variety of fluorescent dyes that may be potentially unsafe but beneficial for imaging. It can be used on any part of human or animal body.