The Colourimetric Method for Mixing Time Measurement in Single-Use and Multi-Use Bioreactors-Methodology Overview and Practical Recommendations

Mixing time is an important parameter for quantifying the mixing efficiency of a bioreactor system, essential for successful bioprocess development in various branches of the bioengineering sector (e.g., biopharma, biorefineries, food industry and bioreactor design). The colourimetric method is one of the ways of obtaining valuable quantitative data about the mixing process and the liquid flow inside a vessel. This review consists of a catalogue and a discussion of previously published data in which the colourimetric method has been utilised; a critical comparison between the colourimetric and other mixing time measurement methods; an explanation of practical considerations regarding the bioreactor setup and the choice of reagents; a practical guide for requirements for the acquisition of high-quality images of the mixing process; a thorough discussion of aspects connected to the computer image processing of the video material. This review is intended to thoroughly present the versatility of the colourimetric method for mixing time measurement in miscellaneous bioreactor systems, i.e., in classical tanks and modern single-use (disposable) plastic film-based bag-like containers, and to facilitate the implementation of the colourimetric method in new research setups by providing complete and valuable recommendations about each step of the methodology.