Green CMFDA

• CAT Number: M116760
• CAS Number: 136832-63-8
• Molecular Formula: C25H17ClO7
• Molecular Weight: 464.85
• Purity: ≥95%
• Synonyms: [6′-acetyloxy-5-(chloromethyl)-3-oxospiro[2-benzofuran-1,9′-xanthene]-3′-yl] acetate
• InChI: InChI=1S/C25H17ClO7/c1-13(27)30-16-4-7-20-22(10-16)32-23-11-17(31-14(2)28)5-8-21(23)25(20)19-6-3-15(12-26)9-18(19)24(29)33-25/h3-11H,12H2,1-2H3
• InChIKey: IPJDHSYCSQAODE-UHFFFAOYSA-N
• SMILES: CC(=O)OC1=CC2=C(C=C1)C3(C4=C(C=C(C=C4)CCl)C(=O)O3)C5=C(O2)C=C(C=C5)OC(=O)C

Green CMFDA is a cell-permeable fluorescent probe with Em of 514 nm and Ex of 485 nm and can be used as a cell tracer. Green CMFDA can be cleaved by non-specific esterases common in living cells, producing a fluorescent compound, fluorescein, visible using a fluorescent microscope[1].
Allow the product to warm to room temperature before breaking seal. Dissolve Green CMFDA in high-quality DMSO to a final concentration of 10 mM as stock solution. Dilute the stock solution to a final working concentration of 0.5-5 µM for short term staining and 5-25 µM for long term staining in serumfree medium. Warm the working solution to 37 ℃.
For cells in suspension:
1. Harvest cells by centrifugation and aspirate the supernatant. Resuspend the cells gently in pre-warmed working solution.
2. Incubate 15–45 min under growth conditions. 3. Centrifuge the cells and remove the working solution.
4. Add culture media and dispense the labeled cell onto slide or into a culture vessel of choice.

For adherent cells:
1. Remove culture media.
2. Gently add the pre-warmed working solution.
3. Incubate 15–45 min under growth conditions.
4. Remove the working solution.
5. Add culture media.

Observe with Emission light of 514 nm and Excitation light of 485 nm.

Reference

[1]. Benjamin J Ross, et al. Challenges in using CellTracker Green on foraminifers that host algal endosymbionts. PeerJ. 2018 Aug 30;6:e5304.
 [Content Brief]