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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 10 — Oct. 1, 2012
  • pp: 2526–2536

Assessment of transferrin recycling by Triplet Lifetime Imaging in living cells

Matthias Geissbuehler, Zuzana Kadlecova, Harm-Anton Klok, and Theo Lasser  »View Author Affiliations

Biomedical Optics Express, Vol. 3, Issue 10, pp. 2526-2536 (2012)

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An optical method is presented that allows the measurement of the triplet lifetime of a fluorescent molecule. This is a characteristic specific to each fluorophore. Based on differences in triplet lifetimes of two fluorescent species (autofluorescence versus label), this novel approach measures relative quantities of a transmembrane receptor and associated fluorescently labeled ligand during its recycling in living cells. Similarly to fluorescence-lifetime based methods, our approach is almost insensitive to photobleaching. A simple theory for unmixing two known triplet lifetimes is presented along with validation of the method by measurements of transferrin recycling in a model system based on chinese hamster ovarian cells (CHO). Transferrin is the delivery carrier for Fe3+ to the cell.

© 2012 OSA

OCIS Codes
(100.2960) Image processing : Image analysis
(170.0180) Medical optics and biotechnology : Microscopy
(170.1420) Medical optics and biotechnology : Biology
(170.3650) Medical optics and biotechnology : Lifetime-based sensing
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.2520) Microscopy : Fluorescence microscopy
(170.2655) Medical optics and biotechnology : Functional monitoring and imaging

ToC Category:

Original Manuscript: July 19, 2012
Revised Manuscript: August 31, 2012
Manuscript Accepted: September 8, 2012
Published: September 13, 2012

Matthias Geissbuehler, Zuzana Kadlecova, Harm-Anton Klok, and Theo Lasser, "Assessment of transferrin recycling by Triplet Lifetime Imaging in living cells," Biomed. Opt. Express 3, 2526-2536 (2012)

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  1. M. Geissbuehler, T. Spielmann, A. Formey, I. Märki, M. Leutenegger, B. Hinz, K. Johnsson, D. Van De Ville, and T. Lasser, “Triplet imaging of oxygen consumption during the contraction of a single smooth muscle cell (A7r5),” Biophys. J.98, 339–349 (2010). [CrossRef] [PubMed]
  2. T. Sandén, G. Persson, P. Thyberg, H. Blom, and J. Widengren, “Monitoring kinetics of highly environment sensitive states of fluorescent molecules by modulated excitation and time-averaged fluorescence intensity recording,” Anal. Chem.79, 3330–3341 (2007). [CrossRef] [PubMed]
  3. W. Rumsey, J. Vanderkooi, and D. Wilson, “Imaging of phosphorescence: A novel method for measuring oxygen distribution in perfused tissue,” Science241, 1649–1651 (1988). [CrossRef] [PubMed]
  4. J. Vanderkooi, G. Maniara, T. Green, and D. Wilson, “An optical method for measurement of dioxygen concentration based upon quenching of phosphorescence,” J. Biol. Chem.262, 5476–5482 (1987). [PubMed]
  5. G. Baravalle, D. Schober, M. Huber, N. Bayer, R. Murphy, and R. Fuchs, “Transferrin recycling and dextran transport to lysosomes is differentially affected by bafilomycin, nocodazole, and low temperature,” Cell. Tissue Res.320, 99–113 (2005). [CrossRef] [PubMed]
  6. H. Li and Z. M. Qian, “Transferrin/transferrin receptor-mediated drug delivery,” Med. Res. Rev.22, 225–250 (2002). [CrossRef] [PubMed]
  7. B. Alberts, Molecular Biology of the Cell (Garland Science - Taylor&Francis group, 2008).
  8. D. Sheff, L. Pelletier, C. O’Connell, G. Warren, and I. Mellman, “Transferrin receptor recycling in the absence of perinuclear recycling endosomes,” J. Cell Biol.156, 797–804 (2002). [CrossRef] [PubMed]
  9. Y. J. Yu, Y. Zhang, M. Kenrick, K. Hoyte, W. Luk, Y. Lu, J. Atwal, J. M. Elliott, S. Prabhu, R. J. Watts, and M. S. Dennis, “Boosting brain uptake of a therapeutic antibody by reducing its affinity for a transcytosis target,” Sci. Transl. Med.3, 84ra44 (2011). [CrossRef] [PubMed]
  10. P. Friden, L. Walus, G. Musso, M. Taylor, B. Malfroy, and R. Starzyk, “Anti-transferrin receptor antibody and antibody-drug conjugates cross the blood-brain barrier,” Proc. Natl. Acad. Sci. USA88, 4771–4775 (1991). [CrossRef] [PubMed]
  11. E. Daro, P. Van Der Sluijs, T. Galli, and I. Mellman, “Rab4 and cellubrevin define different early endosome populations on the pathway of transferrin receptor recycling,” Proc. Natl. Acad. Sci. USA93, 9559–9564 (1996). [CrossRef] [PubMed]
  12. J. Gruenberg and F. Maxfield, “Membrane transport in the endocytic pathway,” Curr. Opin. Cell Biol.7, 552–563 (1995). [CrossRef] [PubMed]
  13. R. Ghosh, D. Gelman, and F. Maxfield, “Quantification of low density lipoprotein and transferrin endocytic sorting in HEp2 cells using confocal microscopy,” J. Cell Sci.107, 2177–2189 (1994). [PubMed]
  14. D. M. Sipe and R. F. Murphy, “High-resolution kinetics of transferrin acidification in BALB/c 3T3 cells: exposure to pH 6 followed by temperature-sensitive alkalinization during recycling,” Proc. Natl. Acad. Sci. USA84, 7119–7123 (1987). [CrossRef] [PubMed]
  15. T. McGraw and F. Maxfield, “Human transferrin receptor internalization is partially dependent upon an aromatic amino acid on the cytoplasmic domain,” Cell Regul.1, 369–377 (1990). [PubMed]
  16. J. R. Lakowicz, Principles of Fluorescence Spectroscopy (Springer, 2006). [CrossRef]
  17. F. Maxfield and T. McGraw, “Endocytic recycling,” Nat. Rev. Mol. Cell Biol.5, 121–132 (2004). [CrossRef] [PubMed]
  18. B. Grant and J. Donaldson, “Pathways and mechanisms of endocytic recycling,” Nat. Rev. Mol. Cell Biol.10, 597–608 (2009). [CrossRef] [PubMed]
  19. A. Durrbach, D. Louvard, and E. Coudrier, “Actin filaments facilitate two steps of endocytosis,” J. Cell Sci.109, 457–465 (1996). [PubMed]
  20. N. Muller, P. Girard, D. Hacker, M. Jordan, and F. Wurm, “Orbital shaker technology for the cultivation of mammalian cells in suspension,” Biotechnol. Bioeng.89, 400–406 (2005). [CrossRef]

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