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Spectroscopy: An International Journal
Volume 27 (2012), Article ID 415496, 5 pages

Study of Cellular Uptake of Modified Oligonucleotides by Using Time-Resolved Microspectrofluorimetry and Florescence Imaging

1Faculty of Mathematics and Physics, Institute of Physics, Charles University in Prague, Ke Karlovu 5, 121 16, Prague 2, Czech Republic
2Laboratoire Jean Perrin, Case Courrier 138, Université P.&M. Curie, 4 Place Jussieu, 75231 Paris cedex 05, France

Copyright © 2012 P. Praus et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Fluorescence microimaging and homodyne phase-resolved confocal microspectrofluorimetry were used to monitor the transport of antisense oligonucleotide into cancer MCF7 cells and the subsequent intracellular distribution. Phosphorothioate analog of 15-mer oligoadenylate (dA15) labeled by ATTO 425 was complexed with 5,10,15,20-tetrakis (1-methyl-4-pyridyl) porphyrin (H2TMPyP4) as an uptake-mediating agent. Fluorescence lifetime data within a broad spectral range have revealed properties of both components inside the cell. H2TMPyP4 lifetime inside the cell is not influenced in this malignant cell line, while the lifetime of modified oligonucleotide was found to be slightly shortened.