Figure 2.
Fibrinogen anchors improves selectivity and homogeneity of micropatterns on PLL-PEG surfaces.(A) NeutrAvidin, NeutrAvidin-DyLight-550, NeutrAvidin mixed with fibrinogen, or NeutrAvidin fused to fibrinogen (Fibrinogen-NeutrAvidin) were micropatterned on PLL-PEG–coated glass using LIMAP with identical UV exposure, micropattern shape, and protein concentration (50 µg/ml). After micropattern quenching and washing, BSA-biotin-Alexa647 (5 µg/ml) was added for 5 min. The sample was then washed, and BSA-biotin-Alexa647 fluorescence was imaged by TIRFM. Two different dynamic ranges for visualization were used for each sample (top versus bottom line) so that each lane could be represented with the same dynamic range. (B) Quantification of the amount of protein bound to the micropattern (left), micropattern selectivity (middle), and homogeneity (right) in the sample presented in C (mean ± SEM). Statistics were performed using a one-way ANOVA test followed by a Tukey post hoc test after log10 transformation of the data (P < 0.001). Fibrinogen-NeutrAvidin enhances significantly the selectivity and the homogeneity of BSA-biotin-Alexa647 micropatterns. (C) GFP and fibrinogen-GFP were micropatterned on PLL-PEG–coated glass using LIMAP with identical UV exposure, micropattern shape, and protein concentration (50 µg/ml). After washing, GFP fluorescence was imaged by TIRFM. (D) Quantification of the amount of protein selectively bound to the micropattern (left), as well as micropattern selectivity (middle) and homogeneity (right) in the sample presented in C (mean± SEM). Statistics were performed using a Student’s t test (after log10 transformation of the data for left and middle panels). The selectivity and homogeneity of fibrinogen-GFP micropatterns are significantly better than those of GFP alone. n, number of micropatterns measured. Scale bars, 10 µm. n.s., not significant.

Fibrinogen anchors improves selectivity and homogeneity of micropatterns on PLL-PEG surfaces. (A) NeutrAvidin, NeutrAvidin-DyLight-550, NeutrAvidin mixed with fibrinogen, or NeutrAvidin fused to fibrinogen (Fibrinogen-NeutrAvidin) were micropatterned on PLL-PEG–coated glass using LIMAP with identical UV exposure, micropattern shape, and protein concentration (50 µg/ml). After micropattern quenching and washing, BSA-biotin-Alexa647 (5 µg/ml) was added for 5 min. The sample was then washed, and BSA-biotin-Alexa647 fluorescence was imaged by TIRFM. Two different dynamic ranges for visualization were used for each sample (top versus bottom line) so that each lane could be represented with the same dynamic range. (B) Quantification of the amount of protein bound to the micropattern (left), micropattern selectivity (middle), and homogeneity (right) in the sample presented in C (mean ± SEM). Statistics were performed using a one-way ANOVA test followed by a Tukey post hoc test after log10 transformation of the data (P < 0.001). Fibrinogen-NeutrAvidin enhances significantly the selectivity and the homogeneity of BSA-biotin-Alexa647 micropatterns. (C) GFP and fibrinogen-GFP were micropatterned on PLL-PEG–coated glass using LIMAP with identical UV exposure, micropattern shape, and protein concentration (50 µg/ml). After washing, GFP fluorescence was imaged by TIRFM. (D) Quantification of the amount of protein selectively bound to the micropattern (left), as well as micropattern selectivity (middle) and homogeneity (right) in the sample presented in C (mean± SEM). Statistics were performed using a Student’s t test (after log10 transformation of the data for left and middle panels). The selectivity and homogeneity of fibrinogen-GFP micropatterns are significantly better than those of GFP alone. n, number of micropatterns measured. Scale bars, 10 µm. n.s., not significant.

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