Journal: bioRxiv
Article Title: mScarlet3-H with low brightness and fluorescence lifetime has potential for cellular lifetime-unmixing and lifetime-based pH-sensing applications
doi: 10.1101/2025.07.15.664898
Figure Lengend Snippet: a. Relative brightness (average ± sd) of mScarlet3 and mScarlet3-H in HeLa cells 24 h after transfection, determined by normalized, spectrally corrected ratios of red to cyanfluorescence intensity found in cells upon co-production of RFP and mTurquoise2. b. maturation speed of mScarlet3-H in HeLa cells measured as delay (average ± sd in min) relative to co-produced mTurquoise2, n=8 cells. c. Photobleaching kinetics of mScarlet and mScarlet3 in living HeLa cells. d. Photobleaching kinetics of mScarlet-H and mScarlet3-H in living HeLa cells. For c. and d. cells were subjected to widefield microscopy and illuminated at ∼ 4W/cm2 with 550/15 nm LED power. Only backgroundfluorescence was subtracted and the netfluorescence intensity was normalized to the initial value at t=0. The average ± sd of four individual cells is shown for each curve.
Article Snippet: After 24 h, fluorescence images were collected using a Nikon Eclipse Ti-E widefield microscope with LEDs at 440 and 555 nm (SpectraX, Lumencor).
Techniques: Transfection, Produced, Microscopy