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Staining of Mitochondria with Cy5-Labeled Oligonucleotides for Long-Term Microscopy Studies

Published online by Cambridge University Press:  09 May 2011

Steffen Lorenz ,
Stephanie Tomcin  and
Volker Mailänder
Steffen Lorenz
Affiliation:
Max-Planck-Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany
Stephanie Tomcin
Affiliation:
Max-Planck-Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany
Volker Mailänder*
Affiliation:
Max-Planck-Institute for Polymer Research, Ackermannweg 10, 55128 Mainz, Germany University Medicine of the Johannes Gutenberg University, III. Medical Clinic (Hematology, Oncology and Pulmonology), Langenbeckstraße 1, 55131 Mainz, Germany
*
Corresponding author. E-mail: volker.mailaender@mpip-mainz.mpg.de
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Abstract

Labeling of organelles for microscopy is achieved generally by specific dyes that either accumulate in a cellular compartment such as cyanine dyes in mitochondria or are only fluorescent under specific conditions such as the low pH in the lysosome. Here we demonstrate that Cy5—a fluorescent molecule that does not enter cells by itself—can be loaded into cells by attaching a short oligonucleotide. This very inexpensive labeling procedure can be done in the presence of serum. Therefore, very sensitive cell types should also be amenable to this procedure, and longer observations can be achieved compared to other commercially available dyes as the labeling reagent does not need to be washed out. This also points to the pitfall of using fluorescently labeled oligonucleotides for live cell imaging where the oligonucleotide is supposed to detect a specific target sequence in its subcellular distribution.

Keywords

Cy5oligonucleotidesmitochondriaCLSM
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 17 , Issue 3 , June 2011 , pp. 440 - 445
Copyright
Copyright © Microscopy Society of America 2011

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References

REFERENCES

Berger, C., Gremlich, H.-U., Schmidt, P., Cannet, C., Kneuer, R., Hiestand, P., Rausch, M. & Rudin, M. (2007). In vivo monitoring the fate of Cy5.5-Tat labelled T lymphocytes by quantitative near-infrared fluorescence imaging during acute brain inflammation in a rat model of experimental autoimmune encephalomyelitis. J Immunol Methods 323(1), 65–77.Google Scholar
Jiang, X., Dausend, J., Hafner, M., Musyanovych, A., Rocker, C., Landfester, K., Mailander, V. & Nienhaus, G.U. (2010). Specific effects of surface amines on polystyrene nanoparticles in their interactions with mesenchymal stem cells. Biomacromolecules 11(3), 748–753.Google Scholar
Lorenz, M.R., Holzapfel, V., Musyanovych, A., Nothelfer, K., Walther, P., Frank, H., Landfester, K., Schrezenmeier, H. & Mailander, V. (2006). Uptake of functionalized, fluorescent-labelled polymeric particles in different cell lines and stem cells. Biomaterials 27(14), 2820–2828.Google Scholar
Pittenger, M.F. (2008). Mesenchymal stem cells from adult bone marrow. Methods Mol Biol 449, 27–44.Google Scholar
Pittenger, M.F. & Martin, B.J. (2004). Mesenchymal stem cells and their potential as cardiac therapeutics. Circ Res 95(1), 9–20.Google Scholar
Poot, M. & Mao, F. (2001). Cyanine dyes that stain cells and mitochondria. U.S. Patent 6,291,203. Eugene, OR: Molecular Probes, Inc.Google Scholar
Rhee, W.J. & Bao, G. (2010). Slow non-specific accumulation of 2′-deoxy and 2′-O-methyl oligonucleotide probes at mitochondria in live cells. Nucl Acids Res 38(9), e109.CrossRefGoogle Scholar
Santangelo, P.J., Nitin, N. & Bao, G. (2005). Direct visualization of mRNA colocalization with mitochondria in living cells using molecular beacons. J Biomed Opt 10(4), 044025.CrossRefGoogle ScholarPubMed
Soltys, B.J. & Gupta, R.S. (1994). Changes in mitochondrial shape and distribution induced by ethacrynic acid and the transient formation of a mitochondrial reticulum. J Cell Physiol 159(2), 281–294.CrossRefGoogle ScholarPubMed
Zhang, C., Liu, T., Su, Y., Luo, S., Zhu, Y., Tan, X., Fan, S., Zhang, L., Zhou, Y., Cheng, T. & Shi, C. (2010). A near-infrared fluorescent heptamethine indocyanine dye with preferential tumor accumulation for in vivo imaging. Biomaterials 31(25), 6612–6617.Google Scholar
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