Acoustic focussing for sedimentation-free high-throughput imaging of microalgae

Microalgae play a key role in aquatic ecology, and methods providing species determination and enumeration can provide critical information about—for instance—harmful algae blooms (HABs) or spreading of invasive species. A crucial step in current methods is the use of sedimentation. This provides the enrichment needed to achieve statistical counts of sometimes rare species within reasonable timeframes, but it comes with the drawback of aggregating the sample. This is a real challenge for computer-aided identification as particle aggregates can often be erroneously classified. In this paper, we propose an alternative method based on flow-through imaging aided by acoustic-focussing, as this provides better input-data for automated counting-methods while simultaneously removing the need for manual sample preparation. We demonstrate that by acoustically focussing microalgae and other particulates in a fast-flowing water sample, it is possible to analyse up to 8 mL sample per minute with sufficient image quality to discriminate the invasive species Ostreopsis ovata from other particulates in samples taken directly from the Mediterranean. We also showcase the ability to achieve sharp images in flow-through at magnifications up to × 50