Optically induced dielectrophoresis for continuous separation and purification of C. vulgaris and H. pluvialis

Marine microalgae are widely present in the natural environment, exhibiting a significant economic value. However, during the inoculation and cultivation process of microalgae, the introduction of unwanted algae is bound to trigger nutrient competition, leading to a decrease in the growth rate of microalgae and consequently impacting their economic value in production. To address this issue, this study integrates the optically induced dielectrophoresis (ODEP) manipulation technology based on the continuous flow in a microfluidic system. A two-stage cell filter, utilizing two virtual optical spots, is designed and manufactured. Leveraging the size differences between microalgae, continuous separation and purification of mixed samples containing Chlorella vulgaris and Haematococcus pluvialis are achieved within microchannels. Additionally, optimal ODEP manipulation conditions for mixed algal liquid samples, comprising C. vulgaris and H. pluvialis, are demonstrated, including appropriate alternating current voltage (6 V), alternating current frequency (100 kHz), light spot width (40 μm), and sample flow rate (0.9 μl/min). Analysis of mixed liquid samples collected at the chip's outlet reveals a reduction in the proportion of H. pluvialis from 37.5% to 1.2% after separation. In summary, this study proposes an ODEP microfluidic system capable of continuously separating and purifying microalgae with different biological characteristics, showcasing its potential as an alternative to traditional labor-intensive microalgae separation techniques.