Worry and FRET: ROS Production Leads to Fluorochrome Tandem Degradation and impairs Interpretation of Flow Cytometric Results
It is hard to overstate the importance of flow cytometry for immunology research. As the field has advanced, the need for an array of fluorophores to meet different excitation and emission characteristics has increased. A predominant method of extending the range of available fluorophores is through addition of tandem conjugates to existing primary fluorophores (e.g., antigen-presenting cell [APC] fluorophore with a Cy7 conjugate yielding APC-Cy7) (Gerstner et al., 2002). This ever-broadening collection of tandem fluorophores has allowed further definition of numerous cell populations and are so often utilized that they are now integral to developing suitable flow cytometry antibody panels, particularly with the advent of modern flow spectral cytometers capable of detecting >30 distinct fluorophores (Cytek, 2020 ).