Tip Box| Infographic
Tips for Optimizing Cell-Based Readouts
Explore five tips for improving detection and reducing background noise with cell-based fluorescence measurements on microplate readers.
 | Red-shifted dyes Cell-derived autofluorescence is mainly found in the green light range. Use red-shifted fluorescent dyes to limit and circumvent nonspecific background fluorescence. |  | Focal height Modulating the microplate reader’s focal plane based on where sample signal intensity is at its highest will ensure optimal detection. For adherent cells grown on microplates, adjust the detection focus of the plate reader to the bottom of the well. |
 | Low-autofluorescence media Cell culture medium components such as serum or phenol red are a source of autofluorescence. Use buffers or a low-autofluorescence medium to reduce this nonspecific background signal. |  | Scan options Plate readers typically measure in the center of the well, which can be problematic when working with heterogeneously distributed adherent cells. Perform well scans that spread measurements across the whole well surface, reducing data variability. |
 | Bottom optics When measuring adherent cells from above the well, excitation and emission light must pass through the potentially auto-fluorescent supernatant. Select bottom optics to avoid autofluorescence effects derived from media or buffer. | Learn more about cell-based fluorescence measurements on microplate readers. |
