How do I minimize cell damage during Fluo-4 AM loading?

Beginning with healthy cells is ideal for any AM ester loading procedure, so that damage incurred during the process is temporary and does not cause any lasting problems. 

Other methods of minimizing cell damage include:

• Ca²⁺indicators are typically dissolved in DMSO stock solution, which is moderately toxic to cells. We suggest mixing the indicator and cell medium first, then adding cells last to minimize damage from DMSO.

• If complex imaging is desired, minimize cellular stress by using a single fluorophore that allows tracking of multiple ions, such as Mag-Fluo-4 AM for Ca²⁺ and Mg ions.