Figure 5
From: Auto-FPFA: An Automated Microscope for Characterizing Genetically Encoded Biosensors
Auto-FPFA can detect structural changes caused by concatemerization. Auto-FPFA was used to characterize and compare Venus monomers (V1, red circles), Venus dimers (V2/V5V, blue squares), Venus tetramers (V4, green diamonds), and Venus hexamers (V6, orange hexagon). Auto-FPFA measurements of molecular brightness (a), diffusion time (τD, b), steady state anisotropy (c), and lifetime (d) of the Venus multimers. Each sample consisted of 5 replicates of 3 different cell homogenates, and black bar is the mean value. The relationship between normalized brightness (e) or fluorescence lifetime corrected normalized brightness (f) is plotted as a function of the number of Venus fluorophores in each construct. Each point is mean ± SD (n = 15). Red line represents a linear fit through the data, and dashed blue lines are the 95% confidence interval.
