Changes of the Microtubule Arrays During Mitosis in Prothallus Cells of Dryopteris crassirhizoma

粗茎鳞毛蕨原叶体细胞有丝分裂过程中微管列阵的变化(英文)

Microtubule arrays in prothalli large-vacuolated and meristematic dividing cells of the fern Dryopteris crassirhizoma Nakai were studied using Steedman's wax, indirect immunofluorescence labelling and confocal laser scanning microscopy. Results showed that the use of high paraformaldehyde concentration (8%) allowed good fixation of prothallus cells, which are characterized by numerous (meristematic cells) and big (large-vacuolated cells) vacuoles. Results also plead for the efficiency of Steedman's wax embedding method in: (1) avoiding excessive use of enzyme for digesting cell wall in the process of the microtubule cytoskeleton labelling, (2) minimizing the autofluorescence effect in cells through utilization of alcohol in sample dehydration, and (3) permitting a clear visualization of microtubule patterns during the cell mitosis. Steedman's wax, coupled with immunofluorescence labelling and confocal laser scanning microscopy techniques, allows a good investigation of cell division process in plants by using simple multicellular organisms such as fern prothalli.

应用Steedman’swax切片法、间接免疫荧光标记技术和激光共聚焦扫描显微镜技术研究了粗茎鳞毛蕨(DryopteriscrassirhizomaNakai)原叶体大液泡化细胞和分生组织细胞有丝分裂过程中微管列阵的变化。结果显示 :应用高浓度的多聚甲醛 (8% )可以很好地保持大液泡化细胞的结构和微管的抗原性。结果也显示Steedman’swax切片法和间接免疫荧光标记技术的优点 :(1)避免在微管标记过程中酶解细胞壁 ;(2 )在乙醇脱水过程中样品中叶绿素的自发荧光被减到最小 ;(3)能够详细观察到有丝分裂过程中微管骨架的变化。因此 ,这种方法可以被广泛用来调查简单植物体和复杂植物体中细胞的有丝分裂过程以及发育过程中微管骨架的变化。