Quantifying Drp1-Mediated Mitochondrial Fission by Immunostaining in Fixed Cells

  • Di Hu
  • Xin QiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2159)


Dynamin-like protein 1 (Drp1) is the master regulator of mitochondrial fission. Drp1 translocates from the cytosol to the mitochondrial outer membrane to execute the scission process. Here we describe an immunofluorescence-based method to measure the mitochondrial translocation of Drp1 and quantify Drp1-related mitochondrial fission by labeling the mitochondrial import receptor subunit TOM20 in fixed cell culture.

Key words

Mitochondrial fission Dynamin-related protein 1 TOM20 Immunofluorescence staining Confocal microscopy 


  1. 1.
    Chan DC (2006) Mitochondria: dynamic organelles in disease, aging, and development. Cell 125:1241–1252. Scholar
  2. 2.
    Chan DC (2006) Mitochondrial fusion and fission in mammals. Annu Rev Cell Dev Biol 22:79–99. Scholar
  3. 3.
    Fannjiang Y, Cheng WC, Lee SJ, Qi B, Pevsner J, McCaffery JM, Hill RB, Basanez G, Hardwick JM (2004) Mitochondrial fission proteins regulate programmed cell death in yeast. Genes Dev 18:2785–2797. Scholar
  4. 4.
    Wakabayashi J, Zhang Z, Wakabayashi N, Tamura Y, Fukaya M, Kensler TW, Iijima M, Sesaki H (2009) The dynamin-related GTPase Drp1 is required for embryonic and brain development in mice. J Cell Biol 186:805–816CrossRefGoogle Scholar
  5. 5.
    Macdonald PJ, Stepanyants N, Mehrotra N, Mears JA, Qi X, Sesaki H, Ramachandran R (2014) A dimeric equilibrium intermediate nucleates Drp1 reassembly on mitochondrial membranes for fission. Mol Biol Cell 25:1905–1915CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Departments of Physiology & BiophysicsCase Western Reserve University School of MedicineClevelandUSA
  2. 2.Center for Mitochondrial DiseasesCase Western Reserve University School of MedicineClevelandUSA

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