Addition of compounds under
investigation can interfere by quenching (underestimation of cell
death) or by auto-fluorescence (overestimation of cell death). To
exclude quenching or auto-fluorescence, one should confirm the
plate reader observations by other non-fluorescent approaches
based on morphologic or biochemical analysis (see Sections 4–6).
The uptake of cell impermeant dyes as a reflection of cell death
can be measured dependent on the available equipment in the first
place, but also on the cell type (cell suspension versus adherent
cells) and the scale of the experiments (small versus high content).
In the section below, we describe in more detail the analysis of the
uptake of cell impermeant dyes by the use of fluorometry, flow
cytometry and high content microscopy.