So every Friday, Rachid teaches a lab class for high schoolers. i.e. is not available for me to pester with questions or to give me instructions. So we got here a little earlier. I ran off to quickly clean off the slides with the plasma cleaner. When I returned, we did a quick test with quantum dots to make sure everything was working. I found them quickly enough and everything looked fine, so Rachid gave me his ideas for the different trials I might do and left. We decided to test the effects that various incubation times of Streptavidin had on the concentration of DNA. So far we had always been doing ten minutes, but in some papers Rachid had read that a lot of people used Streptavidin with a concentration one tenth that of ours or incubated it for only 1-2 minutes.
First I set up my four slides in their holders and gave them all an equal treatment of blocking reagent; after ten minutes, i washed them out and put on some streptavidin. After five minutes i washed out two of them and gave them DNA. For the other two I waited five more minutes before washing out the streptavidin. The first one I looked at; I couldn't see anything. I was worried that it had leaked oto the microscope thus ruining the index of refraction. So i took it off and checked it for leaks. It was fine but the process rempved any chance of us using it for further experiments. The next one came out fine and I took a video. Then I took the first one of the five minute set. I searched probably for close to 15 minutes trying to see something on the slides andwould on occasion see some of the white dots but they were so faint that i assumed they were dust. I think maybe I accidentaly photobleached this slide at one point before putting on the microscope. Then I went to pick up the fourth one but noticed that the paper underneath it was wet, so since it was leaking I just cleaned it up and started preparing four new slides.
This time I kept Streptavidin on two of them for five minutes, and on the other two for only two minutes. I got one video from each type. One of the other ones leaked, and I didn't have time to do the last one.
I converted all the videos to Tiff, saved them to a USB, and went back to my desk to begin the analysis but imageJ shows only static at the beginning followed by white. Im trying to google the problem now but so far no luck.
Also there is an LOB barbeque on Tuesday!
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