Flow cytometry to estimate the cell disruption yield and biomass release of Chlorella sp. during bead milling

Abstract

A number of visual, chemical and fluorescence-based methods are generally employed for monitoring of algae cell growth, culture health and biomass concentration. These methods are often time-consuming, demand destructive and high volume sampling. Rapid, efficient, cost-effective and automated methods which facilitate high-throughput and non-destructive sampling would highly benefit microalgae biotechnology. It is known in literature that with flow cytometry it is possible to monitor microalgae growth and microalgae culture health. Flow cytometry, however, has not been used to estimate biomass release and cell disruption yield. In this study with representative cultures of Chlorella sp., flow cytometry data were generated by a long pass filter (>670nm) and proved to be a promising technique to rapidly evaluate these parameters during cell disruption by bead milling. Both a laboratory and a commercial culture of Chlorella were bead milled. Prior to and during bead milling, biomass release was evaluated gravimetrically and the cell count was determined via hemocytometer manual counting and 3 flow cytometry methods: 1) direct event count of a certain population, 2) quadrant-upper right event count of a long pass filter (>670nm) and 3) a calculation based on the long pass filter data. The data of all methods were compared and correlated to gravimetric biomass release data. Manual counting resulted in an underestimation of the cell disruption yield as one of the Chlorella cultures manual counts did not agree with biomass release data. Good correlations for both cultures were found for cell disruption yield calculations based on flow cytometry data and gravimetric biomass release data. Flow cytometry is therefore an efficient analytical method to rapidly screen disruption yields during cell disruption in microalgae and can be a substitute for the time-consuming direct methods (e.g. gravimetric, manual counting) for estimating biomass release and cell disruption yield.