Department of Physics, Clarkson University, Potsdam, NY 13699, USA.
Skin Res Technol. 2009 Nov;15(4):379-83. doi: 10.1111/j.1600-0846.2009.00369.x.
BACKGROUND/PURPOSE: To 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.
Two 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.
The 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.
The 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.
背景/目的：开发一种改良的剥离方法，以实现体外高分辨率荧光可视化人皮肤的角质细胞。为了验证该方法，从不同年龄的个体中收集了在皮肤薄片上测量的角质细胞面积。
先后使用了两种互补的荧光染料。首先，用阳离子染料（若丹明 640）染色剥离带的粘性层。使用该胶带收集皮肤薄片。然后，用阴离子染料（荧光素）对胶带和收集的薄片进行染色。暴露于第二次染色的胶带的荧光由于染料的相互猝灭而大大降低。13 名健康男性（6-86 岁）参与了该方法的验证。在背部和前臂上进行了测量。
该方法允许通过荧光显微镜对角质细胞进行高分辨率成像。绝对面积以及角质细胞面积与个体年龄的依赖性与先前报道的数据非常吻合。
开发的方法快速且简单。它需要与个体的最小交互作用，并允许使用各种潜在不安全但对成像有益的荧光染料。它可用于人体或动物身体的任何部位。
