One case and the orientation method

A bone marrow aspirate in sodium heparin was processed 5 hours after collection. Sodium heparin and processing within 5 hours meet the locally validated specimen requirements; that status does not establish specimen representativeness or viability. The concurrent morphology preparation has scant spicules, so peripheral-blood admixture remains possible.

The orientation panel uses three linked eight-color tubes. Tube 1 contains CD45, viability dye, CD34, HLA-DR, CD19, CD10, CD20, and surface immunoglobulin. Tube 2 contains CD45, viability dye, CD34, CD19, cytoplasmic CD79a, cytoplasmic MPO, cytoplasmic CD3, and CD10. Tube 3 repeats CD45, viability dye, CD34, CD19, CD10, CD20, surface immunoglobulin, and HLA-DR. CD45, viability dye, CD34, CD19, and CD10 are backbone markers used to locate the same target across tubes; scatter, the sequential gate hierarchy, and those shared backbone markers link tubes rather than an assumed event-by-event match.

The viability method is exclusion of viability-dye-positive events after singlet selection. Specimen retention is different: sodium-heparin collection and five-hour processing meet the retention requirement, whereas viability is the measured exclusion of nonviable events in the acquired specimen. All intensity calls are relative to locally validated controls and internal populations; no universal cutoff applies.