BD Horizon™ Brilliant Stain Buffer

1. Add 50 µL of undiluted BD Horizon™ Brilliant Stain Buffer to each 12 × 75-mm tube.

2. Add the appropriate volume of each fluorochrome-conjugated antibody. NOTE See the antibody reagent IFU for antibody volumes.

3. Vortex to mix well, especially after adding a BD Horizon Brilliant™ reagent.

4. Add 100 µL of the specimen and continue with the staining procedure per the antibody reagent IFU. See the antibody reagent IFU for the staining procedure and storage conditions of stained cells prior to acquisition. If you are staining cells using more than one BD Horizon Brilliant™ Violet reagent in the presence of BD Horizon™ Brilliant Stain Buffer, we recommend acquiring the sample within 6 hours of fixation.

In cases that BD Horizon™ Brilliant Stain Buffer does not adequately resolve DDI, we recommend that you:

- Understand the antigen density of the markers being studied. If DDI is expected, use BD Horizon Brilliant™ Violet reagents for markers expressed at lower levels, if possible.

- Understand the co-expression of markers on the cell types being studied. If dyes exhibiting DDI are used in a panel but the markers are expressed on different cell types and the analysis channels are different for the different cell types, then DDI is not of concern.

- Run trial panels to ensure that when DDI is present, the use of BD Horizon™ Brilliant Stain Buffer minimizes the DDI to such an extent that accurate qualitative decisions can be made. Once the panel has been defined it should then be tested in the presence of bilirubin and hemoglobin to ensure these two potential interferents do not substantively impact the performance of the panel.