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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)
http://dx.doi.org/10.1364/BOE.3.002526


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Abstract

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:
Microscopy

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

Citation
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)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-3-10-2526


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