Tip Box| Infographic
How to Optimize OD600 Measurements
Optical density can be affected by sample conditions, the state of the measuring vessel, and instrument configuration.
 | Well Scanning The presence of bacterial aggregates can increase data variability if measurements are only taken in the center of the well. Well scanning spreads measurement locations across a broader area, thus improving data quality. |  | Condensation Condensation droplets affixed to the lid or sealer can artificially increase optical density measurements. Condensation can be mitigated by not transferring pre-warmed plates to a non-heated microplate reader. |
 | Environment Optimal growth depends on environmental conditions such as temperature and atmospheric composition. Meeting a given organism’s specific requirements encourages growth and reduces cell death and the resultant debris. |  | Path Length Correction When comparing results from different vessels, values must be normalized to the path length of the liquid. Light scattering of OD600 samples negatively affects waterpeak-based methods for path length correction. Use volume-based correction methods instead. |
 | Evaporation Liquid evaporation increases with rising temperature. Insufficient liquid volumes negatively affect OD600 measurements. A gas permeable sealer or plastic lid can help mitigate evaporation. | Learn more about performing OD600 measurements on microplate readers. |
What do you use microplate readers to measure?
