Normal human peripheral blood cells were stained with CD4 Monoclonal Antibody, Alexa Fluor 532 (Product # 58-0049-42), CD19 Monoclonal Antibody, PE-Cyanine7 (Product # 25-0199-42 ), CD8a Monoclonal Antibody, FITC (Product # 11-0087-42), CD25 Monoclonal Antibody, PE (Product # 12-0259-42), CD56 (NCAM) Monoclonal Antibody, PE-eFluor 610 (Product # 61-0566-42) and CD3 Monoclonal Antibody, Alexa Fluor 561. Cells in the lymphocyte gate were used for analysis and first gated on CD3 positive/CD19 negative cells (top left). CD3 positive cells were analyzed for their expression of CD4 and CD8a (top right) and gated on CD4 and CD8a single positive cells, respectively. Expression of CD25 and CD56 (NCAM) was analyzed on CD4 positive cells (bottom left) or CD8a positive cells (bottom right). This data was collected on a 5-laser Cytek Aurora full spectral cytometer. As shown here, Alexa Fluor 561 is compatible with PE and PE tandems when proper panel design principles are applied.