With the expansion of flow cytometry into spectral flow cytometry there is an opening for the development of new fluorophore's with previously under utilised emission peaks. We tested the novel Invitrogen™ NovaFluor™ CD4 Label Characterization Kits using fresh human blood and samples were acquired on three different types of flow cytometry instruments. Once the single stain data was collected the spillover spread for each Fluorophore was calculated using SQI (Spread Quantifcation Index) which allows the comparison between different instruments. We found that there clear resolution could be seen for each NovaFluor antibody and the novel peak emissions could be easily added into already existing multicolour panels.