定量活细胞中的线粒体含量。

Quantifying mitochondrial content in living cells.

作者信息

Viana Matheus Palhares, Lim Swee, Rafelski Susanne M

机构信息

Departments of Developmental and Cell Biology, University of California, Irvine, CA, USA.

Biomedical Engineering, University of California, Irvine, CA, USA.

出版信息

Methods Cell Biol. 2015;125:77-93. doi: 10.1016/bs.mcb.2014.10.003. Epub 2015 Jan 8.

DOI:10.1016/bs.mcb.2014.10.003

PMID:25640425

Abstract

We describe a novel version of MitoGraph, our fully automated image processing method and software, dedicated to calculating the volume of 3D intracellular structures and organelles in live cells. MitoGraph is optimized and validated for quantifying the volume of tubular mitochondrial networks in budding yeast. We therefore include the experimental protocol, microscopy conditions, and software parameters focusing on mitochondria in budding yeast. However, MitoGraph can also be applied to mitochondria in other cell types and possibly other intracellular structures. We begin with our protocol and then include substantial discussion of the validation, requirements, and limits of MitoGraph to aid a wide range of potential users in applying MitoGraph to their data and troubleshooting any potential problems that arise. MitoGraph is freely available at the Web site http://rafelski.com/susanne/MitoGraph.

摘要

我们描述了MitoGraph的一个新版本，这是我们的全自动图像处理方法及软件，专门用于计算活细胞中三维细胞内结构和细胞器的体积。MitoGraph经过优化并经验证，可用于定量分析出芽酵母中管状线粒体网络的体积。因此，我们纳入了侧重于出芽酵母中线粒体的实验方案、显微镜条件和软件参数。不过，MitoGraph也可应用于其他细胞类型中的线粒体以及可能的其他细胞内结构。我们先介绍我们的方案，然后对MitoGraph的验证、要求和局限性进行大量讨论，以帮助广大潜在用户将MitoGraph应用于他们的数据，并排查出现的任何潜在问题。可在网站http://rafelski.com/susanne/MitoGraph上免费获取MitoGraph。