Department of Medical Chemistry, University of Szeged, Szeged, Szikra utca 2, Hungary.
Brain Res Bull. 2011 Oct 10;86(3-4):217-21. doi: 10.1016/j.brainresbull.2011.07.004. Epub 2011 Jul 18.
The cell-impermeant oligomer-(e.g. beta-amyloid-, or tubulin-) specific fluorescent dye, bis-ANS (4,4'-bis-1-anilinonaphtalene-8-sulfonate), was successfully used for labeling mechanically damaged but still viable neuron bodies, neurites and neurite cross sections in acute brain slices. Acute hippocampal brain slices of rats were co-stained with bis-ANS and the cell-impermeant, DNA-specific dye propidium iodide (PI) and were then analyzed using fluorescence and confocal microscopes. Both the neuron bodies and the neurites were found to exhibit increased fluorescence intensities, suggesting that using this method they can be detected more easily. In addition, bis-ANS showed good region - but not cell specific co-localization with the neuron-specific fluorescent dye Dil (1,1'-Dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate). These two dyes label different neuronal structures: Dil binds specifically to intact cell membranes while bis-ANS can enter cells with compromised cell membranes and then stain the microtubules in the cytoplasm. For a quick (10min) staining of acute brain slices with bis-ANS both HEPES and NaHCO(3) were needed in order to achieve high signal intensity. Labeling with bis-ANS fluorescent dye is an easy method for imaging the neuronal structures on the surface of acute brain slices.
不透细胞膜的寡聚物（如β淀粉样蛋白或微管蛋白）特异性荧光染料，双-ANS（4,4'-双-1-苯胺基萘-8-磺酸钠），成功地用于标记机械损伤但仍然存活的神经元体、突起和突起的横切面，在急性脑片中。将大鼠急性海马脑片与不透细胞膜、DNA 特异性染料碘化丙啶（PI）共同染色，然后使用荧光显微镜和共聚焦显微镜进行分析。神经元体和突起都显示出荧光强度增加，表明使用这种方法可以更容易地检测到它们。此外，双-ANS 与神经元特异性荧光染料 Dil（1,1'-十八烷基-3,3,3',3'-四甲基吲哚碳菁高氯酸盐）表现出良好的区域而非细胞特异性共定位。这两种染料标记不同的神经元结构：Dil 特异性结合完整的细胞膜，而双-ANS 可以进入细胞膜受损的细胞，然后在细胞质中染色微管。为了在急性脑片中快速（10min）用双-ANS 进行染色，需要 HEPES 和 NaHCO(3) 才能达到高信号强度。用双-ANS 荧光染料进行标记是一种简单的方法，可以对急性脑片表面的神经元结构进行成像。
