Laser-induced fluorescence of flowing samples as an approach to single-molecule detection in liquids.

A flow cytometer system was used to detect aqueous rhodamine 6G by laser-induced fluorescence. Best results were obtained with careful spectral and spatial filtering. At the detection limit, the probability of a rhodamine 6G molecule being present in the detector's probed volume of 11 pL is about 0.6 . With a flow rate of 0.42 ..mu..L/s, a detection limit of 8.9 x 10/sup -14/ M was obtained for a 1-s time constant. At the detection limit, 18 ag or 22,000 molecules of rhodamine 6G flowed through the probed volume during the signal integration period. Signal linearity extends over greater than 5 orders of magnitude limited only by saturation of the detection electronics at high concentration. The results presented here allow a projection to single-molecule detection with reasonable improvements to the apparatus. 25 references, 5 figures, 7 tables.

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