User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/10/11/Water isotope experiment
9% Heavy Oxygen trial 01
So when I initially started this study, all the microtubules were breaking apart. I'm not sure why this happens but, I let the experiment sit for quite some time between the first run and the sequential runs and now the assay is working. I am very unsure as to why this happens. The antifade that I'm using is greater than a week old but, that has never been a problem up to now so I don't think the problem is an antifade one. It may be that the antifade just didn't mix sufficiently well enough in solution.
Also, when I went to look at the MTs, I noticed that they were really dim. So, I had to bump up the amount of Hg illumination to 50%. Everything seems to be working just fine right with the 50% illumination so I'm going to collect data. If anything, this should be a good way to determine if the temperature I'm at with the objective is sufficient for 50% illumination.
I'm now running the 20th ROI. Due to the odd behavior I was getting early on in the assay, I'm going to let this one run to 30 different ROIs.
After completing the 9% run, I also taught Nadia how to make a PEM solution and the associated tools necessary to make it. We aren't done but, I'm hoping she learns a lot.
81% Heavy Oxygen trial 01
Moving forward to another assay.
I am again having the same issue of catastrophe initially in this assay as well. I'm going to let it sit on the microscope for 10 minutes before I illuminate it again.
It's odd but, waiting seems to work. I waited about 10 minutes and they are still breaking apart. I have taken one ROI and closed the shutter. I'll wait a few more minutes before looking again.
Waiting definitely seems to work. I have no clue why. I was able to collect data and I'm right now tracking it.