BODIPY-Cholesterol (TF-Chol) is a cholesterol analog with a fluorescent BODIPY motif and cellular permeability, and is a novel BODIPY cholesterol probe.BODIPY-Cholesterol can be used to study intracellular sterol uptake and interorganelle sterol flux.

For the BODIPY-cholesterol assay:Reconstitute the analogue in DMSO to prepare a stock solution of BODIPY-cholesterol at a concentration of 1-10 μM[2].
Dilute cells with PBS (or other buffers), wash them (300×g, 10 min) to eliminate excess diluent, and resuspend the loose pellet with vehicle media to achieve the desired concentration[2].Label cell suspensions with BODIPY-cholesterol (final concentration of 0.5-5 μM), ensuring thorough mixing, and incubate for 10-20 minutes at 37 °C[1].Dilute the cells with each of the capacitation media conditions, as outlined in Incubation media, to a final concentration. Incubate for 2 hours before flow cytometric assessment[2].