A VITAL STAIN FOR THE GOLGI-APPARATUS
A VITAL STAIN FOR THE GOLGI-APPARATUS
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DOI：
10.1126/science.2581316
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发表时间：
1985-01-01
期刊：
影响因子：
56.9
通讯作者：
PAGANO, RE
中科院分区：
文献类型：
--
作者：
LIPSKY, NG;PAGANO, RE
高尔基复合体是一种在膜运输和大分子合成中具有重要功能的膜细胞器，在活细胞[中国仓鼠肺成纤维细胞]中用荧光鞘脂染色。首先将细胞与含有N-[7-（4-硝基苯并-2-氧杂-1，3-二唑）]-6-氨基己酰基鞘氨醇（C6-NBD-神经酰胺）的脂质体或与荧光脂质的牛血清白蛋白复合物一起孵育，然后通过荧光显微镜检查。一个强烈的荧光核周结构被确定为高尔基体的共定位与已知的Glogi标记在固定的细胞。C6-NBD-神经酰胺用于观察在存在或不存在莫能菌素或秋水仙胺的活细胞中以及在有丝分裂期间高尔基体的形态。在所有情况下，C6-NBD-神经酰胺揭示了活细胞中的高尔基体，其与用需要固定的常规程序获得的高尔基体相同。
The Golgi complex, a membranous organelle with important functions in membrane traffic and macromolecular synthesis, was stained in living cells [Chinese hamster lung fibroblasts] with a fluorescent sphingolipid. Cells were 1st incubated with liposomes containing N-[7-(4-nitrobenzo-2-oxa-1,3-diazole)]-6-aminocaproyl sphinogsine (C6-NBD-ceramide), or with a bovine serum albumin complex of the fluorescent lipid and then examined by fluorescence microscopy. An intensely fluorescent perinuclear structure was identified as the Golgi apparatus by its colocalization with known Glogi markers in fixed cells. C6-NBD-ceramide was used to observe the morphology of the Golgi apparatus in living cells in the presence or absence of monensin or Colcemid, and during mitosis. In all cases, C6-NBD-ceramide revealed a Golgi apparatus in the living cell that was identical to that obtained with conventional procedures that require fixation.