Unit

UNIT 4.25 Analysis of Mitochondrial Dynamics and Functions Using Imaging Approaches

  1. Kasturi Mitra,
  2. Jennifer Lippincott-Schwartz

Published Online: 1 MAR 2010

DOI: 10.1002/0471143030.cb0425s46

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Mitra, K. and Lippincott-Schwartz, J. 2010. Analysis of Mitochondrial Dynamics and Functions Using Imaging Approaches. Current Protocols in Cell Biology. 46:4.25:4.25.1–4.25.21.

Author Information

  1. Cell Biology and Metabolism Program (CBMP), National Institute of Child Health and Human Development (NICHD), NIH, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAR 2010
  2. Published Print: MAR 2010

Abstract

Mitochondria are organelles that have been primarily known as the powerhouse of the cell. However, recent advances in the field have revealed that mitochondria are also involved in many other cellular activities like lipid modifications, redox balance, calcium balance, and even controlled cell death. These multifunctional organelles are motile and highly dynamic in shapes and forms; the dynamism is brought about by the mitochondria's ability to undergo fission and fusion with each other. Therefore, it is very important to be able to image mitochondrial shape changes to relate to the variety of cellular functions these organelles have to accomplish. The protocols described here will enable researchers to perform steady-state and time-lapse imaging of mitochondria in live cells by using confocal microscopy. High-resolution three-dimensional imaging of mitochondria will not only be helpful in understanding mitochondrial structure in detail but it also could be used to analyze their structural relationships with other organelles in the cell. FRAP (fluorescence recovery after photobleaching) studies can be performed to understand mitochondrial dynamics or dynamics of any mitochondrial molecule within the organelle. The microirradiation assay can be performed to study functional continuity between mitochondria. A protocol for measuring mitochondrial potential has also been included in this unit. In conclusion, the protocols described here will aid the understanding of mitochondrial structure-function relationship. Curr. Protoc. Cell Biol. 46:4.25.1-4.25.21. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • mitochondria;
  • imaging;
  • live cell