Figure 3
From: Pitfalls in optical on-line monitoring for high-throughput screening of microbial systems
In-vitro characterization and correction of the influence of different fluorescent proteins on the optical DOT signal. (A) Dependency of the phase angle as raw signal for DOT monitoring from FbFP, YFP and mCherry fluorescence for DOTs of 100% (closed symbols) and 0% (open symbols) air saturation. (B) Change of DOT calibration curve with varied YFP fluorescence intensity. (C) Change of DOT calibration curve with varied mCherry fluorescence intensity. (D) Correction of the online DOT signal for the cultivation of E. coli expressing YFP by using fluorescence dependent calibration curves. (E) Correction of the online DOT signal for the cultivation of E. coli expressing mCherry by using fluorescence dependent calibration curves. Note: Altered DOT scale in Figures 3D and 3E at higher values. (F) Dependency of the phase angle from mCherry fluorescence for optodes with/without optical isolation (OI). Cultivation conditions: 48well FlowerPlate with optodes for DOT and pH measurement, VL = 800 μL, n = 1100 rpm, d0 = 3 mm, 37°C, Wilms-MOPS medium with 20 g/L glucose, induction with 0.1 mM IPTG after 6 h (indicated by arrow).