Bonaventura Gabriele, Barcellona Maria Luisa, Golfetto Ottavia, Nourse Jamison L, Flanagan Lisa A, Gratton Enrico
Department of Drug Science, Section of Biochemistry, University of Catania, Catania, Italy,
Cell Biochem Biophys. 2014 Nov;70(2):785-94. doi: 10.1007/s12013-014-9982-8.
We describe a method based on fluorescence-lifetime imaging microscopy (FLIM) to assess the fluidity of various membranes in neuronal cells at different stages of development [day 12 (E12) and day 16 (E16) of gestation]. For the FLIM measurements, we use the Laurdan probe which is commonly used to assess membrane water penetration in model and in biological membranes using spectral information. Using the FLIM approach, we build a fluidity scale based on calibration with model systems of different lipid compositions. In neuronal cells, we found a marked difference in fluidity between the internal membranes and the plasma membrane, being the plasma membrane the less fluid. However, we found no significant differences between the two cell groups, E12 and E16. Comparison with NIH3T3 cells shows that the plasma membranes of E12 and E16 cells are significantly more fluid than the plasma membrane of the cancer cells.
我们描述了一种基于荧光寿命成像显微镜（FLIM）的方法，用于评估处于不同发育阶段（妊娠第12天（E12）和第16天（E16））的神经元细胞中各种膜的流动性。对于FLIM测量，我们使用劳丹探针，该探针通常用于利用光谱信息评估模型膜和生物膜中的膜水渗透情况。使用FLIM方法，我们基于对不同脂质组成的模型系统进行校准构建了一个流动性量表。在神经元细胞中，我们发现内膜和质膜之间的流动性存在显著差异，质膜的流动性较小。然而，我们发现E12和E16这两个细胞组之间没有显著差异。与NIH3T3细胞的比较表明，E12和E16细胞的质膜比癌细胞的质膜流动性显著更高。
