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对突变体和药物处理的果蝇胚胎进行实时共聚焦分析。

Live confocal analysis of mutant- and drug-treated Drosophila embryos.

作者信息

Fasulo Barbara, Sullivan William

机构信息

Molecular and Cellular Biology, University of California, Santa Cruz, CA, USA.

出版信息

Methods Mol Biol. 2014;1075:243-55. doi: 10.1007/978-1-60761-847-8_12.

Abstract

The model organism Drosophila melanogaster is particularly well suited for live image analysis. The availability of GFP transgenic flies and a wide array of fluorescent probes, in conjunction with laser scanning confocal microscopy, allow us to image multiple aspects of the cell cycle simultaneously. Confocal microscopy provides the sensitivity and resolution to observe the dynamics of specific cellular events in real time. For example, GFP-histone and rhodamine-labeled tubulin enable one to follow specific nuclear and cytoskeletal events including nuclear envelope formation, nuclear envelope breakdown, spindle formation, centrosome duplication, separation and migration, chromosomes condensation, and segregation. This analysis permits a detailed morphological and temporal description of nuclear and cytoskeletal events in normal or drug-injected embryos.

摘要

模式生物黑腹果蝇特别适合进行活体图像分析。绿色荧光蛋白(GFP)转基因果蝇和各种各样的荧光探针,结合激光扫描共聚焦显微镜,使我们能够同时对细胞周期的多个方面进行成像。共聚焦显微镜提供了实时观察特定细胞事件动态的灵敏度和分辨率。例如,GFP标记的组蛋白和罗丹明标记的微管蛋白使人们能够追踪特定的核和细胞骨架事件,包括核膜形成、核膜破裂、纺锤体形成、中心体复制、分离和迁移、染色体凝聚和分离。这种分析允许对正常或注射药物的胚胎中的核和细胞骨架事件进行详细的形态学和时间描述。

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