The application of flow cytometry for kelp meiospore isolation

The goal of this study was to evaluate flow-cytometric techniques for isolating individual kelp meiospores into 96-welled plates. Previously reported low success rates for sorting tens of thousands of meiospores at a time have been improved by technological developments, specifically, the low nozzle pressure of the commercially available, JSAN (Bay Bioscience Co. Ltd) instrument. We monitored growth and gametophyte development post-sorting for 10 months. Our data demonstrate that successful kelp meiospore isolations of up to 76% viability can be achieved with flow-cytometry. This method can save time as compared to traditional, manual isolations using pipettes and improves confidence that self-fertilized individuals will not contaminate specific crosses of resulting gametophytes. Our results highlight a new application for the flow cytometer to produce clonal kelp gametophytes with direct applications for germplasm and culture collection development.