Create a multicolor flow cytometry experimental workflow: (1) sample preparation (blood/tissue), (2) Fc receptor blocking, (3) surface antibody staining panel design (compensation matrix), (4) fixation/permeabilization for intracellular targets, (5) intracellular antibody staining, (6) instrument setup: laser configuration (405nm, 488nm, 561nm, 640nm) and detector arrangement, (7) compensation bead controls, (8) acquisition with optimized voltages, (9) gating strategy: FSC/SSC → singlets → live/dead → lineage markers → functional markers, (10) FlowJo analysis with t-SNE dimensionality reduction.
