User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/08/07/Passivation study

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Introduction

So I'm going ahead with the belief that keeping the slide cold before putting it on the objective is the way to go. I've also noticed that there is not as much evaporation of material if the slide is on the cooler. Of course this is obvious but I didn't think about it previously.

Whole casein 1st try

  • 1.0mg/mL whole casein in PEM for 10 minutes on the cold plate.
  • 0.5mg/mL whole casein in PEM + 1mM ATP + 27ug/mL kinesin for 5 minutes on the cold plate.
  • Motility solution.

I should point out that it took me longer to get to the first ROI due to improper placement of the slide on the microscope. But, everything seems to be working just fine right now.

  • Movie 2
    • So far there is a mixture of small, medium, and long MTs exhibiting motility. I will note that the focus is drifting a little bit. This means that there is still some temperature gradients going on.
  • Between movies 5 & 6, I had to readjust the slide so that the images were in focus for the entire FOV.

I'm getting some squiggles and some circles reminiscent of the heavy water studies.

Alpha casein 1st try

I really think it is the cold plate that allows me to get passivation. I was able to see motility with the alpha passivation with relative ease. As opposed to when I used the hot plate.

I am having zero problems with this new setup.

  • Movie 3
    • There is a rather odd MT in that the the plus end of it was attached to the kinesin surface but the minus end was in solution. Very odd looking.
  • The focus isn't drifting that much with the alpha casein.

Beta casein 1st try

Moving forward to the next assay.

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