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==Discussion== *Effect of Filter: **It's a good idea to filter the fixed E.coli sample before DAPI staining. The fixed-E.coli suspsend in 1mL PBS can still pass through the 5um filter. **The picture of filtered sample showed much uniformed particles than non filtered samples. **Although it looks like non filtered sample is brighter. However, these bright particles mostly stained large debris. The small particle (probably read E.coli cells) are still dim. *Effect of higher dye concentration: **Although not seen in pictures, it seems like 4uL dye stained sample("F") showed a better signal/noise than 1uL dye staining sample ("R") if we compared under the same microscope setting. **It's effective but result is not significant. *Effect of final washing: **Add finally washing using PBTw decrease the signal intensity significantly, and I can't tell if the background is decreased or not in the washed sample. **I am planing to confirm this again in the next experiment.
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