Merocyanine 540 as a flow cytometric probe of membrane lipid organization in leukocytes.
Merocyanine 540 as a flow cytometric probe of membrane lipid organization in leukocytes.
复制标题
部花青 540 作为白细胞膜脂组织的流式细胞术探针。
DOI：
10.1002/jlb.44.5.337
复制
发表时间：
1988
影响因子：
5.5
通讯作者：
DelBuono,BJ
中科院分区：
文献类型：
--
作者：
McEvoy,L;Schlegel,RA;Williamson,P;DelBuono,BJ
Merocyanine 540 （MC540）是一种荧光探针，优先结合松散包装的脂质膜。当与流式细胞术相结合时，它提供了一种快速定量评估单个白细胞脂质组织的新方法。同时用MC540和1-[4-三甲氨基苯基]-6-苯基-1,3,5-己三烯染色的细胞进行分析，使表面积归一化，发现外周血中所有白细胞每单位表面积结合等量的染料。这一结果表明，所有类型的循环细胞的质膜的脂质组织相似。在适当的刺激下激活后，淋巴细胞、单核细胞和中性粒细胞每单位表面积结合的染料量都增加了，这表明脂质组织向较不有序的状态变化。用MC540染色的细胞根据其荧光强度进行分类，产生相对于脂质组织均匀的群体。因此，不仅可以将MC540和流式细胞术结合起来分析单个白细胞中的脂质组织，还可以获得分选的群体，用于进一步的生化、结构和功能分析。
Merocyanine 540 (MC540) is a fluorescent probe that binds preferentially to membranes with loosely packed lipids. When combined with flow cytometry, it provides a novel methodology for rapidly and quantitatively assessing lipid organization in individual leukocytes. Analysis of cells stained simultaneously with MC540 and 1-[4-trimethylam-moniumphenyl]-6-phenyl-1,3,5-hexatriene, to normalize for surface area, revealed that all leukocytes in peripheral blood bind equivalent amounts of dye per unit surface area. This result indicates that the lipids of the plasma membranes of all types of circulating cells are organized similarly. Upon activation with appropriate stimuli, lymphocytes, monocytes, and neutrophils all bound increased amounts of dye per unit surface area, indicating a change in lipid organization to a less-ordered state. Cells stained with MC540 were sorted on the basis of their fluorescence intensity yielding populations homogeneous with respect to lipid organization. Thus not only can MC540 and flow cytometry be combined for analyzing the organization of lipids in individual leukocytes, but sorted populations can be obtained for further biochemical, structural, and functional analyses.