Hulspas R, Houtsmuller A B, Bauman J G, Nanninga N
Department of Molecular Pathology, Institute for Applied Radiobiology and Immunology TNO, Rijswijk, The Netherlands.
Exp Cell Res. 1994 Nov;215(1):28-32. doi: 10.1006/excr.1994.1310.
Using a beta-tubulin specific antibody, centrosomes were labeled in paraformaldehyde fixed human lymphocytes. Cells were kept in suspension to preserve the three-dimensional (3D) morphology as much as possible. The centrosome was generally identified as the focus of the microtubule array. Resting (G0) and phytohemagglutinin activated cells in G1 stage were taken for 3D analysis of the centrosome position, using confocal microscopy and 3D analysis software. Measurements were performed in relation to the nuclear center and the periphery of the propidium iodide stained area ("nuclear envelope"). The distribution of the distances between the centrosome and the nuclear center revealed that in most resting cells the centrosome was located at the basis of a nuclear indentation. Upon activation, however, the centrosome appeared to move out of the indentation during transition from G0 to G1 stage.
使用β-微管蛋白特异性抗体,在多聚甲醛固定的人淋巴细胞中对中心体进行标记。细胞保持悬浮状态,以尽可能保留三维(3D)形态。中心体通常被确定为微管阵列的焦点。使用共聚焦显微镜和3D分析软件,对处于静止(G0)期和G1期经植物血凝素激活的细胞进行中心体位置的3D分析。测量是相对于碘化丙啶染色区域(“核膜”)的核中心和周边进行的。中心体与核中心之间距离的分布表明,在大多数静止细胞中,中心体位于核凹陷的底部。然而,在激活后,在从G0期向G1期转变过程中,中心体似乎移出了凹陷。