Scheynius A, Dalenbring M, Carlsson K, England R, Lindberg M
Department of Clinical Immunology, Karolinska Hospital, Stockholm, Sweden.
Acta Derm Venereol. 1992 Sep;72(5):348-51.
Confocal laser scanning microscopy (CLSM) was used for quantitative analysis of CD1a+ cells in epidermis at irritant reactions. Sodium lauryl sulphate (2% and 4%) or non-anoic acid (20% and 80%) were applied to the skin of healthy volunteers under occlusion for 24 h. Skin biopsy specimens were taken after additional 24 h and were snap frozen. Freeze-sections, 25 microns thick, were stained with anti-CD1a antibodies (Leu-6) followed by FITC-labelled rabbit anti-mouse IgG. The sections were viewed and optically sectioned in the CLSM at four depth levels. The data was analysed using a threshold value for the fluorescence. The obtained result is presented as the proportion of specimen area having a fluorescence intensity above the threshold. The result demonstrates that the CLSM is a useful tool for obtaining not only structural information but also quantitative information from a defined tissue volume. In the present investigation it was possible to demonstrate variations in CD1a+ reactivity in epidermis at detergent-induced irritant reactions with a marked decrease in CD1a+ after 80% non-anoic acid exposure and only minor differences in the CD1a+ after 2% and 4% sodium lauryl sulphate exposure.
共聚焦激光扫描显微镜（CLSM）用于刺激性反应时表皮中CD1a +细胞的定量分析。将十二烷基硫酸钠（2%和4%）或壬酸（20%和80%）敷于健康志愿者皮肤并封闭24小时。再过24小时后取皮肤活检标本并速冻。制备25微米厚的冰冻切片，用抗CD1a抗体（Leu-6）染色，随后用异硫氰酸荧光素标记的兔抗小鼠IgG染色。在CLSM下于四个深度水平观察切片并进行光学切片。使用荧光阈值分析数据。所得结果表示为荧光强度高于阈值的标本面积比例。结果表明，CLSM不仅是获取结构信息的有用工具，也是从特定组织体积获取定量信息的有用工具。在本研究中，能够证明在洗涤剂引起的刺激性反应中表皮中CD1a +反应性的变化，80%壬酸暴露后CD1a +明显减少，2%和4%十二烷基硫酸钠暴露后CD1a +仅有微小差异。
