title: Time-resolved microspectrofluorometry and fluorescence imaging techniques: study of porphyrin-mediated cellular uptake of oligonucleotides.
abstract: 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.
authors.affiliation: Charles University in Prague, Faculty of Mathematics and Physics, Ke Karlovu 3, Prague 2, CZ-121 16, Czech Republic. praus@karlov.mff.cuni.cz
authors.forename: Petr
authors.initials: P
authors.lastname: Praus
authors.forename: Eva
authors.initials: E
authors.lastname: Kocisová
authors.forename: Peter
authors.initials: P
authors.lastname: Mojzes
authors.forename: Josef
authors.initials: J
authors.lastname: Stepánek
authors.forename: Olivier
authors.initials: O
authors.lastname: Seksek
authors.forename: Franck
authors.initials: F
authors.lastname: Sureau
authors.forename: Pierre-Yves
authors.initials: PY
authors.lastname: Turpin
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002412:Cations; D002467:Cell Nucleus / Q000378:metabolism; D016503:Drug Delivery Systems; D005453:Fluorescence; D008546:Melanoma, Experimental; D051379:Mice; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry*; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D011166:Porphyrins / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D002412:Cations; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D011166:Porphyrins