对活裂殖酵母细胞中的绿色荧光蛋白融合体进行成像。

Imaging green fluorescent protein fusions in living fission yeast cells.

作者信息

Tran P T, Paoletti Anne, Chang Fred

机构信息

University of Pennsylvania, Department of Cell and Developmental Biology, 421 Curie Blvd.-BRB2/3 Room 1009, Philadelphia, PA 19104, USA.

出版信息

Methods. 2004 Jul;33(3):220-5. doi: 10.1016/j.ymeth.2003.11.017.

DOI:10.1016/j.ymeth.2003.11.017

PMID:15157889

Abstract

The use of green fluorescent protein (GFP) fusions as biosensors for examining protein localization and dynamics has revolutionized cell biology. Here, we describe the methods developed for imaging of GFP-fusions in the fission yeast Schizosaccharomyces pombe using fluorescence microscopy, with a focus on the use of time-lapse imaging to analyze the dynamics of microtubules. We discuss the considerations in fluorescence microscopy, cell preparation, data acquisition, and image analysis appropriate for analysis of living cells.

摘要

使用绿色荧光蛋白（GFP）融合体作为生物传感器来检测蛋白质定位和动态变化，彻底改变了细胞生物学。在此，我们描述了利用荧光显微镜对裂殖酵母粟酒裂殖酵母中的GFP融合体进行成像所开发的方法，重点是使用延时成像来分析微管的动态变化。我们讨论了适用于活细胞分析的荧光显微镜、细胞制备、数据采集和图像分析方面的注意事项。