Detection of single molecules of phycoerythrin in hydrodynamically focused flows by laser-induced fluorescence

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Abstract
Single molecules of B-phycoerythrin were detected by laser-induced fluorescence in hydrodynamically focused flows as each molecule transited a focused laser beam. Phycoerythrin is a large protein molecule containing the equivalent of 25 rhodamine-6G chromophores. Single molecule detection is documented by the following: (1) the number of counts per molecules is in agreement with the expected number, (2) the number of molecules per second is in agreement with the concentration and the flow rate, and (3) the time interval distribution between detected molecules is in agreement with the concentration and the flow rate. The molecular transit time through the 1.1-pL probe volume was 180 .mu.s.