Abstract
Photoactivatable caged fluorophores are useful tools for researching the cell system. These caged reagents can be activated on request with UV light as a trigger. The main idea of this research is to activate these reagents by sonolysis instead of photolysis. To activate reagents, we used 1 MHz continuous-wave and burst-wave whose duty ratio was 1:10. For caged reagents, we adopted CMNB-caged fluorescein, dextran conjugates of CMNCBZ-caged carboxy-Q-rhodamine, and CMNB-caged carboxyfluorescein. These caged compounds yield fluorescence after irradiation of UV light. In order to estimate ultrasonic degradation of these reagents, which means releasing the fluorescent group and caging groups, fluorescence intensities were compared the sonicated samples to a control. Sonoactivation of these reagents was done extracellularly and intracellularly. In extracellular experiments, after sonolysis of each sample for 10 seconds to 10 minutes, fluorescence intensity increased with exposure time. Sonochemical activation of reagents in the cultured PLB-985 cells was tested using a flow cytometer and was observed by fluorescence microscope. We found that these reagents were also activatable by ultrasound. Moreover, for more various applications, we used anitibody conjugates of CMNB-caged fluorescein and done experiments in the same way. Our results show it is possible to sonoactivate these caged compounds on behalf of ultraviolet.
