Fig. 3: OLS enables in-solution measurement of protein diffusion. | Nature Methods

Fig. 3: OLS enables in-solution measurement of protein diffusion.

From: Oblique line scan illumination enables expansive, accurate and sensitive single-protein measurements in solution and in living cells

Fig. 3: OLS enables in-solution measurement of protein diffusion.

a, Representative sequential images of isSMT (top) with overlaid tracks (bottom) for JF549-labeled free HaloTag protein at 30% glycerol for a 192 × 192-pixel ROI at 200 Hz. Circles represent the detected PSFs; marks indicate the particle trajectories; color indicates the trajectory ID. Scale bar, 2 μm. b, Mean diffusion coefficient as a function of medium viscosity compared to theoretical Stokes–Einstein equation. 22–44-well replicates with four FOVs per well were acquired for each concentration at 200 frames per second, in duplicate 384-well plates from the same protein preparation. Error bars denote FOV-level s.d. c, Halo-TCF4 diffusion measured with a dose response of either monomeric (monovalent (MV)) versus dimeric (bivalent (BV)) β-catenin protein (Bcat); n = 8 wells. d, Diffusion coefficient of the TCF4–β-catenin complex measured across a dose response of unlabeled TCF4 peptide and toxoflavin; n = 32 FOVs from two distinct plates. Error bars denote the s.d.

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