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酿酒酵母减数分裂的延时荧光显微镜观察。

Time-lapse fluorescence microscopy of Saccharomyces cerevisiae in meiosis.

作者信息

Dresser Michael E

机构信息

Department of Molecular, Cell and Developmental Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA.

出版信息

Methods Mol Biol. 2009;558:65-79. doi: 10.1007/978-1-60761-103-5_5.

Abstract

Movements are implicit in the chromosome behaviors of bouquet formation, pairing and synapsis during meiotic prophase. In S. cerevisiae, the positions of chromosomes, specific structures, and individual chromosomal loci marked by fluorescent fusion proteins are easily visualized in living cells. Time-lapse analyses have revealed rapid and varied chromosome movements throughout meiotic prophase. To facilitate the analysis of these movements, we have developed a simple, inexpensive, and efficient method to prepare sporulating cells for fluorescence microscopy. This method produces a monolayer of cells that progress from meiosis through spore formation, allows visualization of hundreds of cells in a single high-resolution frame and is suitable for most methods of fluorescence microscopy.

摘要

在减数分裂前期,花束形成、配对和联会的染色体行为中隐含着运动。在酿酒酵母中,通过荧光融合蛋白标记的染色体位置、特定结构和单个染色体位点在活细胞中很容易观察到。延时分析揭示了减数分裂前期整个过程中染色体快速且多样的运动。为便于分析这些运动,我们开发了一种简单、廉价且高效的方法,用于制备用于荧光显微镜观察的产孢细胞。该方法产生一层从减数分裂到孢子形成过程的细胞,能在单个高分辨率视野中观察到数百个细胞,并且适用于大多数荧光显微镜观察方法。

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