With positive biochemical and photolysis data in hand we were encouraged to test the caged compounds efficacy under cell culture conditions. Cells were plated on glassbottom chambered coverslips and transiently transfected to express both 5HT2C-GFP fusion protein (Addgene # 79677) and RCaMP-1a (Addgene #61562) genetically encoded calcium indicator. The culture media was loaded with one or both caged compounds. Imaging and uncaging photolysis was performed with a laser scanning confocal microscope. Activation of WinterGreen-WAY or WinterRed-NDMC by laser irradiation occurred at 488 nm or 639 nm, respectively. The 561 nm laser channel was used to observe RCaMP-1a fluorescence before and after irradiation.