Simultaneous metabolic and oxygen imaging is promising to follow up therapy response,dis-ease development and to determine prognostic factors.FLIM of metabolic coenzymes is now widely accepted to be the most reliable ...Simultaneous metabolic and oxygen imaging is promising to follow up therapy response,dis-ease development and to determine prognostic factors.FLIM of metabolic coenzymes is now widely accepted to be the most reliable method to determine cellular bioenergetics.Also,oxygen consumption has to be taken into account to understand treatment responses.The phosphorescence lifetimne of oxygen sensors is able to indicate local oxygen changes.For phosphorescence lifetime imaging(PLIM)dyes based on ruthenium(I)coordination com-plexes are useful,in detaill TLD1433 which possesses a variety of different triplet states,enables complex photochemistry and redox reactions.PLIM is usally reached by two photon exci-tation of the drug with a femtosecond(fs)pulsed Ti:Sapphire laser working at 80 MHz repe-tition rate and(time-correlated single photon counting)(TCSPC)detection electronics.The interesting question was whether it is possible to follow up PLIM 1using faster repetition rates.Faster repetition rates could be advantageous for the induction of specific photochemical reactions because of similar light doses used normally in standard CW light treatments.For this,a default 2p-FLIM-PLIM system was expanded by adding a second fs pulsed laser("helixx")which provides 50 fs pulses at a repetition rate of 250 MHz,more than 2.3 w average power and tunable from 720 nm to 920 nm.The laser beam was coupled into the AOM instead of the default 80 MHz laser.We demonstrated siuccessful applications of the 250 MHz laser for PLIM which correlates well with measurements done by excitation with the conventional 80MHx laser source.展开更多
基金supported by the Ministry of Research and Development,FKZ order:13N14508("OMOXI")by the Ministry of Economics,ZIM-Project,FKZ:ZF4322901RE6("UFEMPU").
文摘Simultaneous metabolic and oxygen imaging is promising to follow up therapy response,dis-ease development and to determine prognostic factors.FLIM of metabolic coenzymes is now widely accepted to be the most reliable method to determine cellular bioenergetics.Also,oxygen consumption has to be taken into account to understand treatment responses.The phosphorescence lifetimne of oxygen sensors is able to indicate local oxygen changes.For phosphorescence lifetime imaging(PLIM)dyes based on ruthenium(I)coordination com-plexes are useful,in detaill TLD1433 which possesses a variety of different triplet states,enables complex photochemistry and redox reactions.PLIM is usally reached by two photon exci-tation of the drug with a femtosecond(fs)pulsed Ti:Sapphire laser working at 80 MHz repe-tition rate and(time-correlated single photon counting)(TCSPC)detection electronics.The interesting question was whether it is possible to follow up PLIM 1using faster repetition rates.Faster repetition rates could be advantageous for the induction of specific photochemical reactions because of similar light doses used normally in standard CW light treatments.For this,a default 2p-FLIM-PLIM system was expanded by adding a second fs pulsed laser("helixx")which provides 50 fs pulses at a repetition rate of 250 MHz,more than 2.3 w average power and tunable from 720 nm to 920 nm.The laser beam was coupled into the AOM instead of the default 80 MHz laser.We demonstrated siuccessful applications of the 250 MHz laser for PLIM which correlates well with measurements done by excitation with the conventional 80MHx laser source.
