Extended data Figure 2: Measuring absolute density of fluorescent molecules on reconstituted target surfaces.
From: Macrophage phagocytosis assay with reconstituted target particles
a, A histogram of fluorescence intensity for four samples of fluorescent beads of known MESF value for Alexa Fluor 488 obtained using flow cytometry. b, Calibration line constructed from plotting the intensity values obtained from flow cytometry and the known MESF value of the beads. The equation for the line of best fit can be used to convert flow cytometry measurements to absolute counts of Alexa Fluor 488 fluorophores, provided that all measurements are performed using the same flow cytometry settings. If the number of fluorophores per molecule of interest (i.e., protein or antibody) is known, along with the surface area of the reconstituted target, an absolute measurement of surface density for the molecule of interest can be obtained.
