摘要 |
The invention relates to a separation free bioanalytical assay method for measuring an analyte from a biological fluid or suspension comprising of microparticles as a bioaffinity binding solid phase, a biospecific secondary reagent labelled with a two-photon fluorescent dipyrrometheneboron difluoride dye, focusing the laser into the reaction suspension measuring two-photon excited fluorescence from single microparticles when they randomly float or are guided by the radiation pressure of the excitation laser through the focal volume of the laser beam using a two-photon fluorescent dipyrrometheneboron difluoride dye. The dye has the structure (II): And at least one of the groups R1, R2, R3, R4, R5, R6 or R7 is substituted to yield a chemically reactive group that can be used for selective covalent linkage to other molecules and at least one of the groups R1, R2, R3, R4, R5, R6, R7 is substituted to yield a water-solubilizing group.
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