Frequency domain fluorescence microspectrometry: Application to cellular uptake and drug distribution
Issue title: From Molecule to Tissue: XIII European Conference on the Spectroscopy of Biological Molecules, Palermo, Italy, August 28–September 2, 2009, Part 2 of 2
Affiliations: Charles University in Prague, Faculty of Mathematics and Physics, Prague, Czech Republic | Acides Nucleiques and Biophotonique, Evry Cedex, France
Note: [] Corresponding author: Petr Praus, Charles University in Prague, Faculty of Mathematics and Physics, Ke Karlovu 3, Prague 2, CZ-121 16, Czech Republic. Tel.: +420 2219 11474; Fax: +420 2492 2797; E-mail: praus@karlov.mff.cuni.cz.
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.
