单细胞LDL受体与清道夫受体活性的同时测定

Simultaneous Determination of LDL and Scavenger receptor Activity in a Single Macrophage.

结合应用激光扫描共聚焦显微镜系统（ＬＳＣＭ）和ＤｉＩＡｃＬＤＬ及ＢＯＤＩＰＹＦＬＬＤＬ两种荧光配基选择性标记技术，可在单细胞水平上同时测定ＬＤＬ受体和清道夫受体活性．Ｃ５７ＢＬ／６Ｊ小鼠巨噬细胞用终浓度为５ｍｇ／Ｌ的ＤｉＩＡｃＬＤＬ及ＢＯＤＩＰＹＦＬＬＤＬ，在３７℃负载５ｈ左右的条件下可获得良好的标记效果．两种荧光配基选择性标记具有高度特异性，在激光共聚焦显微镜下可清晰、定量地观察细胞对ＬＤＬ和ＡｃＬＤＬ摄入。

By combination of laser scanning confocal microscopy(LSCM) and the highly sensitive fluorescent dyes BODIPY FL LDL and DiI AcLDL, it is possible that activities of both LDL and scavenger receptors in a single cell can be measured simultaneously and quantitatively. For example, in C57BL/6J macrophage, it was found that the cells were incubated for 5 h at 37℃ with 5 mg/L DiI AcLDL and 5 mg/L BODIPY FL LDL resulted in excellent color imaging under LSCM, the LDL receptor with green and scavenger receptor with red. The high selectivity and visulization of this method provide detailed information on the localization and activity of both receptors in a single macrophage. The rapidity and accuracy of this assay allows its application for studying receptor mediated lipoprotein uptake.