Burn injuries are a common trauma with life-long consequences including higher rates of cancer and infection years to decades after the injury. Previously, we provided the first evidence of immune dysfunction in paediatric burn survivors >3 years after the injury, including: elevated frequencies of memory and regulatory T-cell subsets, increased circulating inflammatory cytokines (TNF-a, IL-2, IL-7, IFN-γ), and disruption of ordinary vaccine responses. However, the duration and natural history of specific T-cell immunophenotypes and function following injury is unclear. Consequently 15- and 14- parameter flow cytometry panels were designed and optimised for a 5-laser BD LSR Fortessa SORP. The panels were optimised for paediatric cryopreserved peripheral blood mononuclear cells to enable the comprehensive and simultaneous immunophenotyping of distinct T-cell subsets in sequential biobanked samples following burn injury in children. Populations identified by employing our panels include major T-helper subsets (Th1, Th2, Th17, Th9, Tregs; memory vs naïve) and incorporate markers of immunosuppressive phenotype (PD-1, PD-L1), activation (ICOS) and function (granzyme B, IFN-γ).