Begin the quality check of the beads labeled as meso-tetra 4-carboxyphenyl porphine or TCPP beads in the flow cytometer by running and collecting 10, 000 events of the rainbow beads. Then perform a rinse with water and collect 10, 000 events of the TCPP negative beads. After rinsing with water again, collect 10, 000 events of the TCPP positive beads.
Then perform a one minute water rinse before appropriate cleaning and shut down the flow cytometer. A representative outcome of the TCPP bead labeling process determined by flow cytometry is shown here with a standardized profile of rainbow beads. These beads serve as quality checks for standardizing the laser voltages to detect TCPP.
The light scatter profile of amine functionalized polystyrene TCPP labeled beads was observed. The light scatter profile of non-functionalized beads was used as a negative bead control to calculate the compensation matrix. The population indicated by the red rectangle in the TCPP labeled beads was absent in the unlabeled bead suspension representing that the TCPP related particulates were not formed in the absence of TCPP and EDC.
The fluorescence intensity of the labeled beads selected by the black gate was several logs higher than that of the unlabeled beads.
