“The ratiometric reporter HyPer responds to levels of peroxide, such as produced by treatment of the worms with tert-butyl hydroperoxide (tBHP), in the cell by showing an increase in fluorescence when excited at 500 nm and a decrease in fluorescence when excited at 420 nm (A). These changes can be show by graphing the fluorescence measured using ImageJ as the ratio of F500/F420 with increases in the ratio showing an increase in peroxide levels (B).”
The Bioanalytical Pharmacology Core is able to offer ratiometric reporters as a means to examine the physiologic effects of drugs on C. elegans. These reporters exploit the transparent nature of worms to allow the effects to be measured inside of living animals without the need to fix or otherwise disturb the animal. For details please see: Wang et. al. Analyzing cell physiology in C. elegans with fluorescent ratiometric reporters. (2014) Methods 68:508-17. Contact our Bioanalytical Pharmacology Core for more information.