Cell-Tracker Orange CMTMR

Prepare working dye solution

Before opening the dye vial, allow the product to warm to room temperature. Dissolve the lyophilized product in high-quality DMSO to a final concentration of 10 mM. Dilute the stock solution to a final working concentration of 0.5–25 µM in serumfree medium. Warm the CellTracker Working Solution to 37°C.

Note: Avoid amine- and thiol-containing buffers.

Staining protocol for cells in suspension

1. Harvest cells by centrifugation and aspirate the supernatant. Resuspend the cells gently in pre-warmed CellTracker Working Solution (see “Prepare working dye solution“).

2. Incubate 15–45 minutes under growth conditions appropriate for the particular cell type.

3. Centrifuge the cells and remove the CellTracker Working Solution.

4. Add culture media of choice and dispense the labeled cell onto slide or into a culture vessel of choice.

5. Image using the appropriate emission and excitation filters for the CellTracker probe (Ex/Em: 541/565).

Staining protocol for adherent cells

1. Remove culture media.

2. Gently add the pre-warmed CellTracker Working Solution (see “Prepare working dye solution“ ).

3. Incubate 15–45 minutes under growth conditions appropriate for the particular cell type.

4. Remove the CellTracker Working Solution.

5. Add culture media of choice.

6. Image using the appropriate emission and excitation filters for the CellTracker probe (Ex/Em: 541/565).

This protocol only provides a guideline, and should be modified according to your specific needs.