Malínská Kateřina, Jelínková Adriana, Petrášek Jan
Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, Praha 6, 165 02, Czech Republic.
Methods Mol Biol. 2014;1209:1-11. doi: 10.1007/978-1-4939-1420-3_1.
FM (Fei-Mao) styryl dyes are compounds of amphiphilic character that are used for the fluorescence tracking of endocytosis and related processes, i.e., the internalization of membrane vesicles from the plasma membrane (PM) and dynamics of endomembranes. Staining with FM dyes and subsequent microscopical observations could be performed both on the tissue and cellular level. Here, we describe simple procedures for the effective FM dye staining and de-staining in root epidermal cells of Arabidopsis thaliana seedlings and suspension-cultured tobacco cells. The progression of FM dye uptake, reflected by an increased amount of the dye in the endosomal compartments, is monitored under the fluorescence microscope in a time-lapse manner. The data obtained can be used for the characterization of the rate of endocytosis and the function of components of endosomal recycling machinery.
FM（飞毛）苯乙烯基染料是具有两亲性的化合物，用于内吞作用及相关过程的荧光追踪，即质膜（PM）膜泡的内化和内膜动态。用FM染料染色并随后进行显微镜观察可在组织和细胞水平上进行。在此，我们描述了在拟南芥幼苗根表皮细胞和悬浮培养的烟草细胞中进行有效FM染料染色和脱色的简单程序。通过内体区室中染料量的增加反映出FM染料摄取的进程，在荧光显微镜下以延时方式进行监测。所获得的数据可用于表征内吞作用速率和内体循环机制组分的功能。
