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Dipyrromethene difluoride-cholesterol (TopFluor-Cholesterol, TF-Chol) is a widely used cholesterol analogue due to its excellent fluorescence properties and considerable similarity with natural cholesterol in terms of membrane partitioning. However, the suitability of TF-Chol for detecting lysosomal cholesterol deposition has recently been questioned. Here, we highlight the fact that the method of lipid delivery and the analysis of time-point both affect the membrane distribution and labeling pattern of TF-Chol, similarly as with radiolabeled cholesterol. Lysosomal sterol accumulation characteristic to a lysosomal storage disease is most readily detected when the probe is introduced via the physiological route, i.e. as a sterol fatty acid ester in low-density lipoprotein particles. When administered to cells from solvent, lysosomal sterol sequestration becomes evident after an overnight equilibration between membranes.

Original publication




Journal article



Publication Date





1054 - 1057


BODIPY-cholesterol, Niemann-Pick type C disease, TF-cholesterol, cholesterol accumulation, lipid imaging, Biological Transport, Boron Compounds, Carrier Proteins, Cell Line, Cell Membrane, Cholesterol, Esterification, Fibroblasts, Humans, Intracellular Membranes, Lysosomal Storage Diseases, Lysosomes, Membrane Glycoproteins, Microscopy, Fluorescence