DiO (solution) 

DiO (solution) is a long-chain carbocyanine dye. Carbocyanine dyes are widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins^[2].
Solvent and concentration: DMSO: 5 mM

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. Preparing Stain Solutions of DiR
Prepare working solutions: Dilute the stock solutions into a suitable buffer such as serum-free culture medium, HBSS or PBS to make 1 to 5 μM working solutions. We do not recommend storing the aqueous solution for more than one day.
2. Cell staining (Suspension cells)
2.1 Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time. The cell density is 1×10⁶/mL.
2.2 Add 1 mL of DiO working solution, and then incubate at room temperature for 5-30 minutes.
2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
3. Cell staining (Adherent cells)
3.1 Culture adherent cells on sterile coverslips.
3.2 Remove the coverslip from the medium and aspirate excess medium.
3.3 Add 100 μL of working solution, gently shake it to completely cover the cells, and then incubate at room temperature for 5-30 minutes.
3.4 Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

881.72

C₅₃H₈₅ClN₂O₆

34215-57-1

O=Cl(=O)([O-])=O.CCCCCCCCCCCCCCCCCCN1C(C=CC=C2)=C2O/C1=C/C=C/C(OC3=C4C=CC=C3)=[N+]4CCCCCCCCCCCCCCCCCC

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Warren GL, et al. Redistribution of cell membrane probes following contraction-induced injury of mouse soleus muscle.  [Content Brief]

  [2]. Bhowmik BB, et al. Photophysical studies of 3,3' dioctadecyloxacarbocyanine dye in model biological membranes and different solvents. Chem Phys Lipids. 2001 Feb;109(2):175-83.  [Content Brief]

  [3]. Gan WB, et al. Multicolor "DiOlistic" labeling of the nervous system using lipophilic dye combinations. Neuron. 2000 Aug;27(2):219-25.  [Content Brief]

  [4]. Bhowmik BB, et al. Photophysical studies of 3,3' dioctadecyloxacarbocyanine dye in model biological membranes and different solvents. Chem Phys Lipids. 2001 Feb;109(2):175-83.  [Content Brief]