Jurkat Incucyte® NucLight Red Cells are supplied as 1 mL cryopreserved vials (1.5 x 106 cells/mL in 90% FBS and 10% DMSO) containing a stable population of human T lymphocyte cells expressing the Incucyte® NucLight Red fluorescent protein, restricted to the nucleus. Parental Jurkat (clone E6.1) cells (ATCC, Cat# TIB-152) were transduced with the Incucyte NucLight Red Lentivirus Reagent (Cat# 4476; EF1α, puromycin). 48 hours after transduction, the cell population was grown in complete growth media containing 1 µg/ml puromycin, for 3 to 5 days, to select for cells expressing NucLight Red. Following selection the Jukat NucLight Red Cells were validated by comparing morphology and chemotactic migration profiles to the parental cell line. The Jurkat NucLight cells are certified mycoplasma free and are STR authenticated by ATCC. These cells are fully validated for use with the IncuCyte® live-cell imaging and analysis system and are ideal for use in real-time cell counting studies, co-cultures and chemotaxis assays.