Time-resolved microspectrofluorometry and fluorescence imaging techniques: study of porphyrin-mediated cellular uptake of oligonucleotides.
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.
Praus
Petr
P
Charles University in Prague, Faculty of Mathematics and Physics, Ke Karlovu 3, Prague 2, CZ-121 16, Czech Republic. praus@karlov.mff.cuni.cz
Kocisová
Eva
E
Mojzes
Peter
P
Stepánek
Josef
J
Seksek
Olivier
O
Sureau
Franck
F
Turpin
Pierre-Yves
PY
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
D002412:Cations; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D011166:Porphyrins