Hello AAPS Flow community,
As you all know, critical controls for flow cytometry are the single-color allowing to generate the compensation matrix. However, the acquisition defined compensation matrix is not always accurate. I would like to know more about how you recommend handling an inaccurate acquisition defined compensation matrix in order not to report false results but neither to introduce artifact in the data. My questions would be the following:
• If a longitudinal study is performed and the same corrections must be performed through time (for every acquisition), how should it be done and documented?
• If an acquisition defined matrix is inaccurate sporadically, how to handle it? How to correct the matrix and document the action? If FMOs are available, should we correct using FMO and investigate the root cause as it might be coming from the lab (sample handling or lot variation). In the case where no FMO are available, should we correct the matrix so the inter run control behaves as expected?
Also, generating those single-stain and acquiring them is time consuming, hence reusing the same compensation matrix over time can be very useful.
• How frequently do you acquire compensation matrices during a longitudinal study for conventional cytometry? Does that change based on fluorophores included in the panel?
• How frequently do you acquire unmixing controls during a longitudinal study for spectral cytometry? Does that change based on fluorophores included in the panel?
------------------------------
Melissa Mathieu
Senior Scientist Cell Therapy Analytical Development
Turnstone Biologics
Member of the AAPS Flow Cytometry Commitee
Disclaimer: Opinions expressed are solely my own and do not express the views or opinions of my employer.
------------------------------