DiO (DiOC18(3)) is a fluorescent lipophilic tracer and can be widely used to label cells, organelles, liposomes, viruses, and lipoproteins.

Instructions
1. Solution configuration
1. Mother liquor configuration: DiO is usually dissolved with DMSO, and the reserve concentration is 1-5mM.
2. Working solution configuration: DiO solution is diluted with DMEM/pbs/water, and the final concentration is usually 1-10 μM (the specific concentration can be adjusted according to experimental needs).
2. Cell marking
Operation steps:
1. Incubate the cells with DiO solution for a time generally 15-30 minutes (it can also be optimized according to the needs of the cell type).
2. Wash cells with PBS buffer to remove unbound DiO.
3. Observe the cells using a fluorescence microscope (usually green fluorescence, excitation of about 480-500 nm).
2. Organelle marking
Operation steps:
1. Prepare the DiO solution and add it to the cell culture medium, usually at a concentration of 1-5 μM.
2. Incubate for 15-30 min under cell culture conditions.
3. Wash cells with PBS to remove unbound dye.
4. Use a fluorescence microscope or flow cytometry to observe the fluorescent label of the organelles.
3. Liposome and lipoprotein markers
Operation steps
1. Mix the DiO solution with liposomes or lipoproteins at appropriate concentrations and incubate.
2. Remove unbound dyes in an appropriate manner.
3. Observe the distribution of markers by fluorescence microscopy or other imaging techniques.