Assessment of transferrin recycling by Triplet Lifetime Imaging in living cells

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.


Published in:
Biomedical Optics Express, 3, 10, 2526-2536
Year:
2012
Publisher:
Washington, Optical Soc Amer
ISSN:
2156-7085
Keywords:
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 Record created 2012-09-13, last modified 2018-03-18

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