Potential of BODIPY-cholesterol for analysis of cholesterol transport and diffusion in living cells

Imaging approaches for analysis of cholesterol distribution and dynamics in the plasma membrane

“…1) partitions to about 60% in the lo-like phase, which is very similar to KK114-PEG-Chol and 3-C6-NBD-Chol. In cells, the transport of BChol and B-P-Chol is comparable to that of DHE and these BODIPY-tagged cholesterol analogues accumulate in cholesterol-rich organelles, as the ERC [73,76]. Extending the linker between the BODIPY-group and cholesterol's side chain, however, alters the phase partition of such probes in model membranes and gives analogues which redistribute to many intracellular membranes different from cholesterol [76].…”

“…The extinction coefficient and quantum yield of such BODIPY-tagged cholesterol probes are very high, and the BODIPY moiety shows low environmental sensitivity, making these probes useful for cellular studies on cholesterol trafficking [73]. However, the incorporation of BODIPY in the cholesterol molecule has some impact on its properties, in particular in living cells.…”

“…One of the best known and most widespread types of fluorescent dyes is bora-diaza-indacene (BODIPY), and BODIPY-tagged cholesterol analogs as TopFluor-cholesterol (or BChol) and the related B-P-Chol are useful probes to track cholesterol in cells by fluorescence microscopy [73].…”

“…Further discussion of the properties and applications of various fluorescent cholesterol analogues is beyond the scope of this article but can be found elsewhere [73,[87][88][89].…”

A comparative study on fluorescent cholesterol analogs as versatile cellular reporters

“…A decrease in D for large excitation powers indicates enhanced photobleaching, i.e., low photostability, because the fl uorescent molecules turn dark prior leaving the observation spot ( 30 ). Here, all analogs except the NBD-Chol showed a satisfactory performance for excitation powers of up to 40 W, while in the case of NBD-Chol, values of transit times had already dropped off at values of 5-10 W. In a nutshell, only TF-Chol, Star512-PEG-Chol, and especially KK114-PEG-Chol were suitable for FCS measurements, while fi lipin III, dansyl-Chol, and the NBD-labeled probes For DHE and TF-Chol, and also for Chol, it has been shown that part of the sterol transport between the plasma membrane and the ERC takes place by a nonvesicular mechanism ( 67,68 ). This transport mode requires fast trans-bilayer migration of the sterol, and indeed, the majority of DHE and CTL has been shown to reside after membrane labeling on the inner leafl et of the plasma membrane ( 69 ).…”