AAT Bioquest

Why are my cells leaking Fluo-4 AM?

Posted February 19, 2020


Fluo-4 AM typically has reasonable cell retention-if you are experiencing unexpected leakage of the indicator, we recommend initial troubleshooting as follows:


  • Check that your protocol is optimized for the selected cell type.
  • Lower temperatures will mitigate cell leakage, or adding probenicid to the media. Both can negatively effect results, however, so employ either method with caution.


If very high cell retention is required, consider Fluo-8® AM as an alternative fluorophore.

Additional resources

Fluo-4 AM

Calcium Indicators

Screen Quest™ Fluo-4 No Wash Calcium Assay Kit