Fig. 3

Visualization of sfGFP(N39D/A179A) expression in P. thermoglucosidasius DSM 2542 at different temperatures. Cells were grown to mid-exponential growth phase in TGP broth and the intensity of sfGFP(N39D/A179A) GFP emission in 50,000 single cells was analyzed by flow cytometry (FC). SSC-A denotes the side scatter area and equivalents the granularity of each detected cell; FL-1(530/20)-A denotes the fluorescence channel 1 area at 530/20 nm excitation, which equals the fluorescence intensity of each detected cell. The cells with fluorescence intensities within the range of the autofluorescence of wild-type DSM 2542 are colored red (gate P1). The cells with a fluorescence signal above the autofluorescence level of DSM 2542 are colored green (gate P2). Highly fluorescent cells are shown as blue dots in gate P3. The threshold between P2 and P3 was chosen arbitrarily. Fluorescence microscopy images were acquired at 100× magnification and at 475/28 nm excitation for 0.3 s and 523/36 nm emission. Representative FC histograms and microscopy images from three independent biological replicates are shown