Journal: Nature chemical biology
Article Title: Acoustic Biosensors for Ultrasound Imaging of Enzyme Activity
doi: 10.1038/s41589-020-0591-0
Figure Lengend Snippet: (a) Coomassie-stained SDS-PAGE gel of OD500nm-matched samples of GVWT incubated with dTEV and TEV protease, before and after buoyancy purification (labeled pre b.p. and post b.p., respectively). N = 3 biological replicates. (b) Scatter plots showing normalized OD500nm of GVSTEV as a function of hydrostatic pressure. (N = 3 biological replicates for GVSTEV + TEV and N =4 for GVSTEV + dTEV.) (c) Scatter plots showing the ratio of nonlinear (x-AM) to linear (B-mode) ultrasound signal as a function of applied acoustic pressure for all the replicate samples used in the x-AM voltage ramp imaging experiments for GVSTEV. N = 3 biological replicates and total number of replicates is 8. (d) Scatter plots showing normalized OD500nm of GVWT as a function of hydrostatic pressure. (N = 3 biological replicates for GVWT +dTEV and N=4 for GVWT + TEV.) (e) Representative ultrasound images of agarose phantoms containing GVWT incubated with TEV or dTEV protease at OD500nm 2.2. The B-mode image was acquired at 132kPa and the x-AM image at 569 kPa. Similar images acquired for N=3 biological replicates, with each N consisting of 3 technical replicates. CNR stands for contrast-to-noise-ratio, and color bars represent relative ultrasound signal intensity on the dB scale. Scale bars represent 1 mm (f) Scatter plots showing the ratio of nonlinear (x-AM) to linear (B-mode) ultrasound signal as a function of applied acoustic pressure for all the replicate samples used in the x-AM voltage ramp imaging experiments for GVWT. N=3 biological replicates, with each N consisting of 3 technical replicates. Solid curve represents the mean of all the replicates.
Article Snippet: B-mode images were acquired at a transmit voltage of 1.6V (132 kPa), and an automated voltage ramp imaging script (programmed in MATLAB) was used to sequentially toggle between B-mode and x-AM acquisitions.
Techniques: Activity Assay, Staining, SDS Page, Incubation, Purification, Labeling, Imaging
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