User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2009/09/10/Surface passivation studies
To keep moving forward, I'm going to keep writing my "To do" list at the top of each notebook.
- Hydrophobic surface.
- Hydrophobic surface with a groove.
- Corn starch.
- Potato starch.
- Silica nanoparticles.
- Silanized surface. I'm not sure how to do this one but apparently lots of people do it to attache DNA to a coverslip.
- Lipids with PEG on a hydrophobic surface spin coated.
- Better spin coating of lipids on coverslips.
- PE lipids?
- ATP study
Koch talked about how to clean slides and how he used the patented ultra awesome Windex method. It is designed to clean glass isn't it? At any rate, I had a buddy of mine here at CHTM also clean us some slips and slides using the ridiculous overkill Piranha method. I will try both versions to see if one is better than the other.
Koch and I also came to the conclusion that the 100 µL aliquots of 100 mM ATP we have is too much to be storing in the e•IceBuckets for extended periods of time. So, Brian and I made 5 µL aliquots.
I tried both the Piranha and ethanol cleaned slides to no avail. There is simply no motility. This could mean one of 2 things now; the kinesin is bad or the casein is bad. To find out if the casein is bad, I am going to way overload the flow cell with kinesin. That way what ever denaturing occurs to the kinesin on the slide will make a bed for some to be active on. If, this doesn't work, then I know it is the kinesin which has gone south.
Andy Maloney 21:53, 10 September 2009 (EDT): I believe that the kinesin is bad. The reason I belive this is because I put a crazy amount of it on a slide and I only saw a minimal amount of motility.
Steve Koch 22:23, 10 September 2009 (EDT): How long ago did you thaw the kinesin aliquot? Do you have the message from Haiqing that I think I forwarded? I can dig it up, but I think she only used it for max 2 weeks after thawing. And in any case, it's definitely not very stable at 4C (my theory is because it aggregates and denatures). Hopefully you were using an oldly-thawed aliquot, because that's an easy fix! (For fun, you could look at the old kinesin under DIC microscopy and see if you see the same aggregates (beads on a string) that I saw (I can dig up my images someday).)