High throughput single-cell sorting for secretion assay studies

The Flow Cytometry Unit at the Life Sciences & Engineering (LS&E) Infrastructure Center applies various instrumentation for cell analysis and sorting. The unit enables diverse analysis applications: from flow imager (Image Stream), through high throughput analysis using LSR, to spectral analysis using the Cytek Aurora Flow analyzer.

For sorting – the unit operates three sorters: BD FACS Aria, BD Influx, and a new high throughput cell sorter – BigFoot, which enables rapid sorting into tubes, or fast single-cell sorting.

High throughput single-cell sorting allows rapid sorting of a single cell into various multi-well plate types (96, 384, 1534) in seconds.

Here, we present the use of the BigFoot sorter for studying secretion factors by S. Cerevisiae cells.

The first goal was to determine the minimal number of cells required for recovery after sorting. We found that recovery and outgrowth from even a single cell are possible. In the second experiment, we created mixed samples of wild-type (WT) and EGFP-expressing cells, with a known ratio. The goal was to determine the minimal ratio detectable in a real mixed sample. We sorted 100 cells of each mixed sample into a single well and found that the ratio detection limit is EGFP:WT approximately 10:90.

We found that High Throughput single-cell sorting preserves high cell viability, and can be applied for further analysis applications, such as detection of secretion factors, genomics, and proteomics.