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LD540 is a novel high-sensitivity lipophilic dye modified with BODIPY fluorescent groups, designed for precise labelling and imaging of lipid droplets. LD540 exhibits excellent photostability and an optimal fluorescence spectrum, making it compatible with various commonly used fluorescent dyes (such as DAPI and Alexa Fluor 647), thereby supporting multi-colour imaging requirements. Additionally, LD540 is suitable for both fixed and live cells and can label ultra-small lipid droplets.
LD540 exhibits virtually no fluorescence in water and other polar solvents, but emits bright red fluorescence when bound to neutral fats such as triglycerides. In ethanol, LD540 has a maximum excitation wavelength of approximately 537 nm and a maximum emission wavelength of approximately 544 nm.
Staining of cells with LD540
Fixed cells
For a stock solution, LD540 was dissolved in ethanol at 0.5 mg/mL, which was stored at −20°C. Cells were fixed with 4% paraformaldehyde, permeabilized with saponin and immunostained with antibodies as described previously. Cells were incubated with a solution of 0.05–0.1 μg/mL LD540 in PBS for 5–30 min, washed three times with PBS and once with water and mounted in Mowiol/Dabco as described. For some applications, it can be useful to avoid glycerol-containing mounting media such as Moviol, because they were reported to induce LD fusion. When antibody staining was not necessary, cells were only fixed without permeabilization and stained with LD540 as described above.
Living cells
Normal growth medium was exchanged for DMEM (without phenol red), 25 mm HEPES pH 7.4 supplemented with 10% fetal calf serum (FCS) (live cell imaging medium, LCIM). Cells were stained with LD540 at 0.5 μg/mL in LCIM for 10 min followed by three washes with LCIM. The higher concentration of LD540 needed is probably due to the presence of large amounts of BSA in the FCS, which captures some of the dye.
https://pubmed.ncbi.nlm.nih.gov/19765264/
| Molecular Weight | 314.19 |
| Formula | C18H21BF2N2 |
| CAS Number | 151486-58-7 |
| Solubility (25°C) | Ethanol |
| Storage | -20°C, protect from light |
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