(A) The carboxylated blank red and orange fluorescent beads were activated.
(B) The free TM target was coupled onto activated carboxylated beads by EDC/NHS to prepare bead probe set as the beadeTM compound.
(C)The TM on the bead and the free TM were allowed to compete for their corresponding mAb in solutions; then, the beadeTMemAb compound was formed.
(D)Sec-Ab- biotin was added to capture them Ab in solutions and to result in the beadeTMemAbesec-Ab-biotin compound.
(E) Streptavidin-R-phycoerythrin (SA-PE) was added into the solutions to couple with the compound by biotin-avidin interaction. The beadeTMemAbesec-Ab-biotineSA-PE compound was ready for laser beaming. The red laser (Ex1) was used to excite the red and orange fluorescent dyes within the beads to recognize their unique coded numbers. Simultaneously, a green laser (Ex2) was used to excite SA-PE that combined on the bead surface. The available fluorescent signalsdMFIs that only combined on the beads were recorded and analyzed by xPONENTâ„¢ 3.0 software. Reprinted from [96] With permission. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)