Comparison: Motility of actin filaments before adding EDTA Vs. after adding EDTA in channel 2 which was applied with EDTA

As shown in this figure, in the experiment, we injected EDTA into the channel 2 to explore the effect of EDTA on the motility of actin filaments.

After we got the video showing the motility of actin filaments before EDTA and after EDTA recorded by EMCCD camera, we use ImageJ to analyze the motility of these actin filaments. By using imageJ, we have traced the moving of actin filaments and obtained some numerical analysis results.

Figure 1: Upper image showing the initial state of actin filaments in channel2 before applying EDTA; Lower image showing the initial state of actin filaments in this channel after applying EDTA

Figure 1 indicates that after we applied EDTA, most of actin filaments were killed by EDTA.

The following images show the tracing result of the motility of actin filaments before applying EDTA and after applying EDTA, the blue tails indicate that the tracked filaments are moving leftward; the yellow tails indicate that the tracked filaments are moving rightward.

Figure 2: Upper image shows the motility of actin filaments at 2.01s before EDTA;

Lower image shows the motility of actin filaments at 2.01s after EDTA

Figure 3: Upper image showing the motility of actin filaments at 3.01s before applying EDTA;

Lower image showing the motility of actin filaments at 3.01s after applying EDTA;

Figure 4: Upper image showing the motility of actin filaments at 4.01s before applying EDTA;

Lower image showing the motility of actin filaments at 4.01s after applying EDTA;

By observing the above tracing results, it can be found that EDTA can stop the movement of most actin filaments, only few of these actin filaments survived after applying EDTA.

Comparison of the numerical analysis results obtained:

Figure 5: Difference tracking results before EDTA

Figure 6: Difference tracing results after EDTA

By comparing the moving intensity, the moving intensity of actin filaments before applying EDTA is about 91372 which is much higher than the moving intensity 323 after EDTA. In addition, by comparing the data of moving count, it can easily find that the number of moving actin filaments is much smaller after applying EDTA. The numerical results identically correspond to the results obtained from the above fluorsecent images.