植物学报
植物學報
식물학보
ACTA BOTANICA SINICA
2003年
2期
193-199
,共7页
微管%分生细胞%大液泡化细胞%有丝分裂%Steedman's wax 切片法%激光共聚焦扫描显微镜术%粗茎鳞毛蕨
微管%分生細胞%大液泡化細胞%有絲分裂%Steedman's wax 切片法%激光共聚焦掃描顯微鏡術%粗莖鱗毛蕨
미관%분생세포%대액포화세포%유사분렬%Steedman's wax 절편법%격광공취초소묘현미경술%조경린모궐
microtubule%meristematic cell%large vacuolated cells%mitosis%Steedman's wax sectioning%confocal laser scanning microscopy%Dryopteris crassirhizoma
应用Steedman's wax 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(Dryopteris crassirhizoma Nakai)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's wax 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.
應用Steedman's wax 切片法、間接免疫熒光標記技術和激光共聚焦掃描顯微鏡技術研究瞭粗莖鱗毛蕨(Dryopteris crassirhizoma Nakai)原葉體大液泡化細胞和分生組織細胞有絲分裂過程中微管列陣的變化.結果顯示:應用高濃度的多聚甲醛(8%)可以很好地保持大液泡化細胞的結構和微管的抗原性.結果也顯示Steedman's wax 切片法和間接免疫熒光標記技術的優點:(1)避免在微管標記過程中酶解細胞壁;(2)在乙醇脫水過程中樣品中葉綠素的自髮熒光被減到最小;(3)能夠詳細觀察到有絲分裂過程中微管骨架的變化.因此,這種方法可以被廣汎用來調查簡單植物體和複雜植物體中細胞的有絲分裂過程以及髮育過程中微管骨架的變化.
응용Steedman's wax 절편법、간접면역형광표기기술화격광공취초소묘현미경기술연구료조경린모궐(Dryopteris crassirhizoma Nakai)원협체대액포화세포화분생조직세포유사분렬과정중미관렬진적변화.결과현시:응용고농도적다취갑철(8%)가이흔호지보지대액포화세포적결구화미관적항원성.결과야현시Steedman's wax 절편법화간접면역형광표기기술적우점:(1)피면재미관표기과정중매해세포벽;(2)재을순탈수과정중양품중협록소적자발형광피감도최소;(3)능구상세관찰도유사분렬과정중미관골가적변화.인차,저충방법가이피엄범용래조사간단식물체화복잡식물체중세포적유사분렬과정이급발육과정중미관골가적변화.
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.
