Syntheticbiology: Next generation genetically encoded fluorescent sensors for serotonin NatureComms
). Imaging was performed 24 h post-transfection. For this, the culture medium was replaced by PBS and 100 µM of NPEC-caged-serotonin was added to the dish. HEK293T cells expressing the 5-HT variants were imaged on a laser scanning confocal Zeiss LSM 880. We used the 488 nm laser for imaging the fluorescence of the 5-HT sensor variants. Imaging was performed using ×60 oil immersion objective at an imaging speed of 0.93 Hz.
Videos were recorded with a CMOS camera at a framerate of 1 Hz. To evaluate if changes in pH alone can elicit fluorescence changes of the sensor, we washed in modified Ringer buffer (140 mM NaCl, 3.5 mM KCl, 0.5 mM NaH
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