Progress achieved in the stability and precision of flow cytometers as well as in analysis software permits the acquisition of reliable data in any competent laboratory. Due to the sophistication and versatility of the new instruments allowing for multi-color analyzes, and due to the large number of reagents available, it has been necessary to define the most pertinent strategies to achieve results that would be comparable between several laboratories.
In the Harmonemia group*, 17 laboratories from 11 countries on 3 continents cooperated with aim to assess the feasibility of a simple, easy to perform and robust method to harmonize settings between instruments of different manufacturers in different laboratories. We found that instruments harmonization could be performed based on reference channels for PMT settings (target channels). Local beads can be later assessed on any harmonized platform to provide adapted target channels. Ongoing harmonization can be performed locally by checking new bead batches against previous settings. Unstained blood provides a good and cheap control of satisfactory PMT settings. Compensations can be applied to any panel using the same fluorochromes. The outline of normality can be assessed by merging large numbers of normal samples. An example of technology transfer of a complex flow cytometry panel between two laboratories using the Harmonemia concept will be presented.
* Lacombe F, et al. Harmonemia: a universal strategy for flow cytometry immunophenotyping - A European LeukemiaNet WP10 study. Leukemia. 2016 Aug;30(8):1769-72