Currently, bioreactors are only sampled once per day or less due to the limitations of labor, bioreactor volume, etc. Daily manual sampling means data unavailable for hours, delaying troubleshooting efforts and identification of problems. The iLine F microscope from Ovizio utilizes Differential Digital Holographic Microscopy (DDHM)- a new quantitative imaging technique that can monitor cell counts and viability in-line at real-time without staining. The microscope captures holograms of the cells travelling through the BioConnect disposable fluidics system, which connects the microscope to the bioreactor. The closed loop set-up minimizes contamination during sampling without sample volume consumption.<br/>In this study, the iLine F microscope was tested against conventional cell quantitation methods such as using flow cytometry (different flourescence staining dyes), trypan blue dye exclusion and capacitance probe measurement. While the trypan blue dye exclusion is the most common method for cell quantitation and viability measurements, it tends to report higher viability when compared to fluorescence staining techniques such as by propidium iodide [1]. The capacitance probe utilizes frequency-dependent polarization of dielectric material to detect live cells by measuring capacitance [2]. Capacitance probes have limitations that they cannot detect non-viable cells. In addition, these probes measure biovolume that must be correlated to viable cell concentration which is subject to change over the culture duration. The iLine F microscope, on the other hand, is based on holographic microscopy and can detect at the single cell level. The study shows proof of concept of monitoring cell growth and viability in real-time as well as the performance comparison with other methods. Different cell lines were also tested to confirm the feasibility and robustness.

Cell characterization using a novel online cell quantitation microscopic technology in comparison with conventional offline methodologies

255th National Meeting (2018)

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