Analyzing membrane dynamics with live cell fluorescence microscopy with a focus on yeast mitochondria

Methods Mol Biol. 2013:1033:275-83. doi: 10.1007/978-1-62703-487-6_17.

Abstract

With the availability of increasing numbers of fluorescent protein variants and state-of-the-art imaging techniques, live cell microscopy has become a standard procedure in modern cell biology. Fluorescent markers are used to visualize the dynamic processes that take place in living cells, including the behavior of membrane-bound organelles. Here, we provide two examples of how we analyze the membrane dynamics of mitochondria in living yeast cells using wide field and confocal microscopy: (1) Long-term observation of mitochondrial shape changes using mitochondria-targeted fluorescent proteins and (2) monitoring the behavior of individual mitochondria using a mitochondria-targeted version of a photoconvertible fluorescent protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Regulation, Fungal
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Fluorescence* / methods
  • Mitochondrial Membranes / metabolism*
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*

Substances

  • Luminescent Proteins