User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/11/02/Water isotope study

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36% O-PEM 1st try

This is the first experiment where I am using 36% heavy oxygen PEM.

Last time I tried to run experiments, I couldn't get microtubules to polymerize. Now I can with a new TSB solution.

I'm getting some breakage in the first microtubules that I have observed. This may be due to the AF although, I made some new antifade this morning. I have noticed before that the slower the microtubules move, they are more prone to catastrophe. This could explain why when I'm close to the edges of the flow cell, the microtubules tend to break up more readily.

In the second ROI, there hasn't been breakage yet and I'm half way through the observation. And, finishing the ROI with no catastrophes.

45% O-PEM 1st try

Again, this assay is moving without a hitch. Seems like it won't break up in the beginning ROIs as well.

Yep. Zero breakups in the first ROI.

9% O-PEM 2nd try

The first ROI in this assay broke apart. The second ROI seems to be looking good.

Yep, no breakups in this one.

72% O-PEM 1st try

No problems here. I did have to step away from the experiment when I took the 1st ROI. I'm not sure how long I was gone but, everything is working just fine.

18% O-PEM 1st try

This assay started without a hitch. Everything looks great.

54% O-PEM 1st try

I'm getting break ups in the 1st ROI for this assay. It was bad so I had to move the FOV to see if there are MTs that are stable. In the new FOV, I do see MTs but they are starting to break apart. I hope the next area doesn't do this.

In the second ROI, I see excellent movement of the MTs. This means that they are at a lower chance of breaking apart. We shall see.

Yep, no more problems are occurring.

27% O-PEM 2nd try

This assay seems to have started without any hitches. There are breakups in the first ROI however. I moved the FOV to look at other MTs. They seem to be moving better.

In the second ROI, everything is moving well and I don't think there will be any more breakups in this assay.

81% 2nd try

Continuing forward. There doesn't seem to be any problems initially with this assay.

18% 2nd try

Again, no problems with the start of this assay.

63% 2nd try

I had to mix up some more O-PEM. I'll have to write up how I did it later.

No problems with this assay. At least, until things started breaking apart. Oh well. We shall see what happens with the next ROI.

Things are definitely moving better in the second ROI. I still have no clue as to why sometimes the first ROI has breaking MTs in it.

63% 3rd try

Just for fun, I ran the 63% O-PEM with the same motility solution I made for the second try. The first ROI again has MTs that are breaking up.

I moved the first ROI to some other MTs and these guys seem to be moving better. With no breakage.