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Spectroscopy
Volume 24 (2010), Issue 3-4, Pages 303-307
http://dx.doi.org/10.3233/SPE-2010-0436

Frequency domain fluorescence microspectrometry: Application to cellular uptake and drug distribution

Petr Praus,1,3 Eva Kočišová,1 Peter Mojzeš,1 Josef Štĕpánek,1 Franck Sureau,2 and Pierre-Yves Turpin2

1Charles University in Prague, Faculty of Mathematics and Physics, Prague, Czech Republic
2Acides Nucleiques and Biophotonique, Evry Cedex, France
3Charles University in Prague, Faculty of Mathematics and Physics, Ke Karlovu 3, Prague 2, CZ-121 16, Czech Republic

Copyright © 2010 Hindawi Publishing Corporation. 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.

Abstract

Time-resolved confocal microspectrofluorometry and fluorescence microimaging were used to monitor how the model antisense oligonucleotide is transported into 3T3 living cells and distributed inside them. Phosphorothioate analog of 15-mer oligothymidylate labeled by ATTO 425 was complexed with Zn(II) 5,10,15,20-tetrakis(4-N-methylpyridyl) porphyrin as an uptake-mediating agent. Homodyne phase-resolved technique based on a high frequency analog modulation of both exciting diode laser and detector image intensifier was used for time-resolved measurements. Decay-time data obtained within a broad range spectral region have provided unique information about the fate of both fluorophores inside the cell.