Establishment of a simple method to evaluate mixing times in a plastic bag photobioreactor using image processing based on freeware tools

OBJECTIVE: Accurate determination of the mixing time in bioreactors is essential for the optimization of the productivity of bioprocesses. The aim of this work was to develop a simple optical method to determine the mixing time in a photobioreactor. The image processing method should be based on freeware tools, should not require programming skills, and thus could be used in education within high schools and in early stages of undergraduate programs. RESULTS: An optical method has been established to analyze images from recorded videos of mixing experiments. The steps are: 1. Extraction of a sequence of images from the video file; 2. Cropping of the pictures; 3. Background removal; and 4. Image analysis and mixing time evaluation based on quantification of pixel-to-pixel heterogeneity within a given area of interest. The novel method was generally able to track the dependency between aeration rate and mixing time within the investigated photobioreactor. In direct comparison, a pearson correlation coefficient of rho = 0.99 was obtained. Gas flow rates between 10 L h(−1), and 300 L h(−1) resulted from mixing times of between 48 and 14 s, respectively. This technique is applicable without programming skills and can be used in education with inexperienced user groups.