Which components of cell culture media can cause background fluorescence?

Short answer: Common components such as phenol red and riboflavin can contribute to background fluorescence and reduce imaging quality in live cell imaging.

Components Causing Background Fluorescence

There are several standard components of cell culture media that can interfere with the live imaging of your cells. Phenol red, often found in DMEM, is used as a pH indicator in cell culture. However, this compound can quench the signal of some visible wavelength fluorescent dyes during live cell fluorescence microscopy.

Another component of DMEM is riboflavin, which is autofluorescent and can negatively affect the signal-to-noise ratio.

How to Reduce Background Fluorescence

If your dye signal is strong, the background created by phenol red and other compounds is usually not critical. However, if your fluorescent signal is low, we recommend using a phenol red-free medium or a specialized medium for live cell imaging.

Additional recommendations for optimizing your imaging workflow can be found in the cell preparation for imaging guide.