High throughput flow cytometry

Background: Conventional flow cytometry does not allow the rapid analysis o f multiple samples. This has limited its uses in drug discovery, for which the standard for throughput is 100,000 samples per day. Methods: We describe a simple method in which com mercial peristaltic tubin g is connected from a commercial autosampler to a flow cytometer. The sampl es are delivered via a peristaltic pump from source wells in a multiwell pl ate. The samples are separated by air bubbles. Results: Throughput rates approach the limit of the autosampler (up to 100 wells per minute). Using optimal tubing and flow rates, particles remain wi thin appropriate light scatter and fluorescence gates. The carryover betwee n wells is typically less than 5% without and 1% with a wash step. The volu mes of sample delivered are in the microliter scale. The approach has been validated with instruments from three manufacturers. Conclusions: Flow cytometry has potential throughput of 100,000 samples or more per day starting with the method described. The method is currently be st suited to end-point assays. However, combined with high-speed sorting an d single- cell assays, the number of assays could approach 1 billion per da y. Cytometry 44.83-90, 2001. (C) 2001 Wiley-Liss. Inc.