User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/03/08/Fixed microtubules

Fixed microtubules...Attempt 1

In order to fix microtubules to a glass slides, we use poly-L-lysine. A previous attempt at getting fixed microtubules failed due to bad Taxol and possibly due to using too much poly-L-lysine.

Planning for another run

Scheduling with Emmalee will be determined either here or through email. The things that I can do on my end include:

• Preparing fluorescein microtubules.
• Determining the correct amount of poly-L-lysine to use as our "fixer".
• Ensuring that we can fix microtubules reproducibly.

Experiment

Emmalee's Notes:
In the past weeks, I have made lipid vesicles using both straight PC lipid and a combination of PC and PG with Texas Red that I will use for the experiments with MTs. I was able to use dynamic light scattering to find the size of the vesicles to be around 100 nm. (Hydronamic radius varied from 50-70 nm) Unfortunately, while I was using the mini-extruder to make my last batch of vesicles, the end of one of the syringes broke off. I have ordered a replacement, and these are the steps I would like to take once it arrives.

• Visualize MT and vesicles on polylysine surface (I can bring over a Texas Red cube for the microscope)

• Introduce tau onto surface with MT and vesicles and observe any change in binding to MT

• Use physiological and mutant hyperphosphorylated tau

I asked for rush shipping on the syringe, so hopefully it will get here before the end of the week. After it arrives, I will need a few hours to prepare new vesicles and confirm their size with DLS.

I anticipate being able to work with the vesicles Friday (Mar 12). At the latest, I should have them ready the next Monday (Mar 15).