C6 NBD Ceramide is a cell-permeable Golgi fluorescent probe (Ex=466 nm, Em=536 nm), a neuraminidic acid with a green fluorescent tag that can recognize Golgi apparatus in live and fixed cells, observing morphological changes of the Golgi apparatus in live cells. C6 NBD Ceramide can be metabolized intracellularly to generate fluorescent sphingomyelin and glucosylceramide, thus applied in studies of sphingolipid transport and metabolism mechanisms.

The recommended protocol for labeling the Golgi apparatus [3] is as follows: Begin by rinsing cells on glass cover slips with HMEM and fixing them in glutaraldehyde or paraformaldehyde for 5-10 minutes at 25℃. Then, wash the fixed cells using a balanced salt solution like HCMF. Transfer the cells in their culture dishes to an ice-water bath and treat with freshly prepared NaBH4 for three 5-minute intervals if fixed with glutaraldehyde. Subsequently, rinse the cells multiple times in cold HCMF for 20-30 minutes, warm them to 25℃, and incubate with C6 NBD-ceramide/BSA for 60 minutes. Following incubation, wash the cells with HCMF and treat with either 10% fetal calf serum or 2 mg/mL BSA for 30-90 minutes at 25℃ to facilitate back-exchange of excess C6 NBD-ceramide and enhance Golgi staining. Finally, mount the glass cover slips on depression slides and examine under a fluorescence microscope with an excitation of 466 nm and emission of 536 nm.