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Time‐resolved Microspectrofluorometry and Fluorescence Imaging Techniques: Study of Porphyrin‐mediated Cellular Uptake of Oligonucleotides
Author(s) -
Praus Petr,
Kočišová Eva,
Mojzeš Peter,
Štěpánek Josef,
Seksek Olivier,
Sureau Franck,
Turpin PierreYves
Publication year - 2008
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1430.007
Subject(s) - porphyrin , fluorescence , confocal microscopy , confocal , fluorescence lifetime imaging microscopy , fluorescence microscope , biophysics , oligonucleotide , chemistry , microscopy , fluorescence correlation spectroscopy , photochemistry , dna , microbiology and biotechnology , biochemistry , biology , optics , physics
Time‐resolved confocal microspectrofluorometry and fluorescence microscopy imaging were applied to monitor the cellular uptake of fluorescent‐labeled oligonucleotides (ONs) delivered by a porphyrin molecule. The fate of porphyrin–ON complexes inside living cells has also been monitored. Due to intrinsic fluorescence of the porphyrin and sensitivity of its characteristics to microenvironment, multicomponent analysis of time‐resolved fluorescence provides unique information about stability of the porphyrin–ON complexes, ON interactions with their target sequences, and ON and porphyrin distributions after delivery inside the cells. Time‐resolved confocal microspectrofluorometry indeed delivers additional information compared with fluorescence confocal microscopy imaging widely employed to study ON uptake.