The word “validation” strikes fear and anxiety in the hearts of many researchers crossing into the world of flow cytometry for clinical trials. Validation plans evaluate assay reliability and overall performance and may include but are not limited to measuring precision, robustness, sample stability, assay specificity, and intersample variability.
Many research scientists working in basic research settings haven’t had to consider how to translate a basic flow cytometry protocol into a validated protocol and may think this is such an overwhelming process that they should avoid clinical trial research altogether. However, any well-trained scientist can develop a validated flow cytometry assay, especially with the assistance or collaboration with scientists who have validation expertise.
Consider these three factors when you are confronted with the need for a validated assay and alleviate your validation fears.
1. Give Yourself Time.
Validation is customizable to any flow cytometry protocol, but some protocols are more complicated depending on the cell type being used, the sample source, or the overall complexity of the assay itself. Give yourself several weeks to develop a validated assay, so you won’t totally derail your work when you hit roadblocks, such as discovering you can only reliably use fresh instead of cryopreserved samples, which could in turn alter your entire study design.
2. Delve Into Documentation.
A hallmark of assay validation is thorough documentation, so it can meet the correct standards needed for clinical trials or diagnostic use. The degree of documentation is dramatically different for scientists who are only familiar with conducting basic research. Be sure to find reliable resources and expert advisors to help you determine the appropriate documentation needed for validation.
3. Be Confident In The Test Scripts.
Some scientists think that a quasi-quantitative assay like flow cytometry cannot be validated, but validation experts have demonstrated repeatedly that flow cytometry assays are able to be validated when appropriate test scripts are used. Depending on your needs, you may only need to use a few specific test scripts instead of doing a full blown validation. It’s all dependent on the level of confidence you need for your assay.
Flow cytometry assays can be validated and is within your reach. Don’t let distress of validation derail your next project!