Figure 6
From: Auto-FPFA: An Automated Microscope for Characterizing Genetically Encoded Biosensors
Auto-FPFA characterization of Venus-Twitch4-mCherry, a hetero-FRET based calcium biosensor. Auto-FPFA was used to measure count rate (a), <N> (b), brightness (c), diffusion time (τD, d), steady state anisotropy (e), and average lifetime (f) of two different homogenates prepared from cells expressing Venus-Twitch4-mCherry. Homogenates were diluted into buffers with defined calcium concentrations where pCa indicates the log of the free calcium concentration in nano-moles. Each point is the mean ± SD (N = 4, two from each homogenate preparations). Dashed lines are a four parameter, variable slope dose-response fit.
