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Analysis of mitochondria isolated from single cells

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Abstract

Bulk studies are not suitable to describe and study cell-to-cell variation, which is of high importance in biological processes such as embryogenesis, tissue differentiation, and disease. Previously, capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was used to measure the properties of organelles isolated from millions of cells. As such, these bulk measurements reported average properties for the organelles of cell populations. Similar measurements for organelles released from single cells would be highly relevant to describe the subcellular variations among cells. Toward this goal, here we introduce an approach to analyze the mitochondria released from single mammalian cells. Osteosarcoma 143B cells are labeled with either the fluorescent mitochondrion-specific 10-N-nonyl acridine orange (NAO) or via expression of the fluorescent protein DsRed2. Subsequently, a single cell is introduced into the CE-LIF capillary where the organelles are released by a combined treatment of digitonin and trypsin. After this treatment, an electric field is applied and the released organelles electromigrate toward the LIF detector. From an electropherogram, the number of detected events per cell, their individual electrophoretic mobilities, and their individual fluorescence intensities are calculated. The results obtained from DsRed2 labeling, which is retained in intact mitochondria, and NAO labeling, which labels all mitochondria, are the basis for discussion of the strengths and limitations of this single-cell approach.

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Acknowledgements

This work has been supported through grant AG20866; KJO was supported through GM08347, and EAA through K02-AG21453.

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Author notes

  1. Ryan D. Johnson

    Present address: Environmental Health Sciences Division, School of Public Health, 140 Warren Hall, University of California Berkeley, Berkeley, CA, 94720, USA

  2. Guohua Xiong

    Present address: US Environmental Protection Agency, National Exposure Research Laboratory – Exposure and Dose Research Branch, 944 Harmon Ave, Las Vegas, NV, 89119, USA

  3. Hossein Ahmadzadeh

    Present address: Department of Chemistry, California State Polytechnic, Pomona, CA, 91768, USA

  4. Ciarán F. Duffy

    Present address: Abbott Ireland Diagnostics Division, Lisnamuck, Longford, Ireland

Authors and Affiliations

  1. Department of Chemistry, University of Minnesota, Minneapolis, MN, 55455, USA

    Marian Navratil, Bobby G. Poe, Dmitry Andreyev & Edgar A. Arriaga

  2. Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN, 55455, USA

    Karen J. Olson & Edgar A. Arriaga

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  1. Ryan D. Johnson
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  2. Marian Navratil
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Corresponding author

Correspondence to Edgar A. Arriaga.

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Johnson, R.D., Navratil, M., Poe, B.G. et al. Analysis of mitochondria isolated from single cells. Anal Bioanal Chem 387, 107–118 (2007). https://doi.org/10.1007/s00216-006-0689-6

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  • DOI: https://doi.org/10.1007/s00216-006-0689-6

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