Automated, high-throughput immunofluorescence staining: a new approach

A high-throughput antibody staining and washing system for large-scale routine indirect immunofluorescence testing was developed, using a robotic sample processor. The Tecan robotic sample processor (Miniprep 75/2, Tecan Benelux BV, Giessen, The Netherlands) was introduced, adapted and evaluated for its suitability for large-scale immunofluorescence screening of potato samples for the presence of the plant pathogenic quarantine bacterium, Ralstonia solanacearum. Throughput and quality of antibody-antigen reactions were compared to manual sample preparation for the immunofluorescence test according to a standard EU testing protocol, using 216 standard potato extracts, including 204 known negative extracts and 12 known positive extracts, and 24 positive control strain samples. Throughput of the Tecan robotic sample processor easily outcompeted manual staining of immunofluorescence slides. The robot was twice as fast as manual processing and is easily cost effective. With the robot 288 samples per day can be processed as compared to 144 samples per day with manual processing. No difference between quality of antibody-antigen reactions and readability of preparations under a microscope could be detected. The new Tecan robotic sample processor was found to be useful for high-throughput fluorescent antibody staining and washing in large-scale, routine indirect immunofluorescence testing.