User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2011/01/25/Reusable flow cell

Initial trial

This trial will consist of me using:

• The 100W Hg lamp.
• 1.0 mg/mL alpha casein passivation. 10 minute incubation.
• Fluid exchange with 0.5 mg/mL alpha casein with 27μg/mL kinesin and 1 mM ATP. 5 minute incubation using twice as much fluid to passivate.
• Fluid exchange with regular motility solution using 20 μL.

I will take data for about 20 minutes or 10 data points. This allows me to get the slide to the objective temperature with enough stable data points to be able to determine the average speed. Once I have taken this data, the slide will come off of the microscope and a fluid exchange with the same regular motility solution will occur. Again, 20 μL of fluid and 10 data points. I am going to repeat this for a total of 9 times or until I see something catastrophic occur in the data taking.

The 9 times is because there will be 9 different solutions of fluid exchange for the water isotope study. Hopefully my fluid exchanges will not affect the passivation and kinesin adhesion on the slide. But, this is what needs to be determined.

Andy Maloney 12:35, 25 January 2011 (EST): So far there has been no hang ups with the 1st ROI. I will have to move the slide a bit however because it doesn't seem to be totally in focus.

First washing

Andy Maloney 13:03, 25 January 2011 (EST): So far everything looks to be working just fine. I did see one issue and that is the objective oil. I'll just have to be careful when changing the fluid so that no oil gets into the chamber. I also decided that I will toss the vinyl strips used to "seal" the flow cell between each fluid exchange.