The analysis of AgNPs Characterizations by TEM.

<p>A. The AgNPs show the regular nanoparticle shape with the diameter ranging from 5 to 10nm. B. The elemental silver was conformed by EDX spectrum during the TEM experiment.</p

The observation of the micrograph of <i>E</i>. <i>coli</i> cells by TEM.

<p>A. Control group with no AgNPs stimulated. The typical morphology of <i>E</i>. <i>coli</i> was observed with the integrity cell membrane. B. <i>E</i>. <i>coli</i> stimulated with 5 μg/ml AgNPs group. The integrity cell membrane with no significant obvious damage can be found. Besides that, several electron dense granules also can be observed (the red arrow). C. <i>E</i>. <i>coli</i> stimulated with 10 μg/ml AgNPs group. There are also electron dense granules in <i>E</i>. <i>coli</i> (the red arrow), and the cell membrane also show the integrity.</p

The analysis of dead ratio in bacterial cells by flow cytoometric.

<p>A. Flow cytometric analysis for dead bacterial percentage using PI staining. B. Flow cytometric analysis control: negative control using bacteria shaken 1h without any treatment and positive control using bacteria co-cultured with 70% isopropanol. C. statistic analysis by one way ANOVA test and Tukey’s multiple comparison test. (n = 3).</p

The new DNA systhesis analysis by ELISA.

<p>A. growth inhibition results of different concentration of AgNPs at the same time. B. Growth inhibition for the same concentration of AgNPs at different times. All data was analyzed by One-way ANOVA test and Tukey’s multiple comparison test following (n = 3).</p

The analysis of the apoptosis in the cells.

<p>A. Flow cytometric analysis for apoptosis bacterial percentage using Annexin V-PI staining (one experiment results). B, C and D. statistic analysis for apoptosis bacterial percentage by One-way ANOVA test, followed Tukey’s multiple comparison test (n = 3). B shows the total percentage of apoptosis bacteria. C shows the percentage of apoptosis bacteria in early stage, including the bacteria in the PI-negative Annexin V-positive quadrant. D shows the percentage of apoptosis bacteria in middle and late stage, including the bacteria in PI-positive Annexin V-positive quadrant.</p

Determination of <i>E</i>. <i>Coli</i>. growth by taking the absorbance at 600 nm and counting CFU.

<p>A. The normal growth curve by analyzing the absorbance at 600 nm. B. The normal growth curve by counting the CFU. C. The assessment of toxicity of NP-Ag in <i>E</i>. <i>Coli</i>. growth by plotting the curve of absorbance. D. The toxicity of NP-Ag in <i>E</i>. <i>Coli</i>. growth by CFU counting.</p