Skip to content

Understanding the 3 rules of compensation, and applying them to your everyday workflows, is an essential step in good, consistent, and reproducible flow cytometry data. Making sure the controls are bright, and treated the same way, is essential. Don’t bring unfixed controls when your samples are fixed, as the controls will not reflect the spectra from the fixed samples. Make sure not to rely on the “Universal Negative”, use a single sample to set background, and collect enough events to make sure an accurate measurement is made, as this will further improve the quality of your control and therefore the data.

Read More

3 different theories on compensation are discussed. The first, non-pensaton, is not recommended, and only possible under a narrowly defined instrument. The second, manual compensation, is also not recommended for anything more than 2 fluorochromes. It is error prone and subject to the researcher’s judgement, unless statistics are invoked and then it becomes a tedious and difficult exercise in algebra. For polychromatic flow cytometry, best practices in flow cytometry is to use the automated compensation methodologies. This will ensure consistent and accurate compensation, if some rules are followed.

Read More