Open AccessCentrifugation‐Free Magnetic Isolation of Functional Mitochondria Using Paramagnetic Iron Oxide Nanoparticles
Author(s) -
Banik Bhabatosh,
Dhar Shanta
Publication year - 2018
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/cpcb.26
Subject(s) - mitochondrion , centrifugation , organelle , differential centrifugation , isolation (microbiology) , cell fractionation , chemistry , biophysics , microbiology and biotechnology , biochemistry , biology , bioinformatics , enzyme
Subcellular fractionation techniques are essential for cell biology and drug development studies. The emergence of organelle‐targeted nanoparticle (NP) platforms necessitates the isolation of target organelles to study drug delivery and activity. Mitochondria‐targeted NPs have attracted the attention of researchers around the globe, since mitochondrial dysfunctions can cause a wide range of diseases. Conventional mitochondria isolation methods involve high‐speed centrifugation. The problem with high‐speed centrifugation‐based isolation of NP‐loaded mitochondria is that NPs can pellet even if they are not bound to mitochondria. We report development of a mitochondria‐targeted paramagnetic iron oxide nanoparticle, Mito‐magneto, that enables isolation of mitochondria under the influence of a magnetic field. Isolation of mitochondria using Mito‐magneto eliminates artifacts typically associated with centrifugation‐based isolation of NP‐loaded mitochondria, thus producing intact, pure, and respiration‐active mitochondria. © 2017 by John Wiley & Sons, Inc.