Figure 2.

Automated Microfluidics Feedback Control Enables Precise Regulation of α-Synuclein Expression over Time in Both WT α-Synuclein Strain and A53T Mutant Strain

α-Synuclein-GFP fluorescence was quantified in each cell and normalized to the red fluorescence (mCherry protein) with a custom-made image processing algorithm (STAR Methods). Light gray lines show representative examples of single-cell traces (STAR Methods). Single-cell traces may start or end at different times, because of cells being born and cells being pushed out of the field of view.

(A and B) Population-averaged α-synuclein-GFP fluorescence (blue with SD across cells in gray) for the WT α-synuclein strain (A) and the A53T mutant α-synuclein strain (B) grown in the microfluidics device in the presence of galactose. Examples of images below each time course show WT and A53T mutant α-synuclein-GFP at the indicated time points.

(C and D) Population-averaged α-synuclein-GFP fluorescence (blue with SD in gray) was controlled to the reference target value (yellow) by automatically switching between glucose and galactose (brown) as computed in real time by the Model Predictive Control strategy (STAR Methods). No α-synuclein inclusions were observed over the course of the experiment in both strains. Examples of images below each time course show (C) WT and (D) A53T mutant α-synuclein-GFP at the indicated time points.

See also Figure S1.