- Finish decanting microspheres.
- Rinse and vacuum filter microspheres with hexanes.
The general protocol described on 2013/03/01 was followed.
- It was found that the organic safflower oil layer needed to be removed several times. After removing a layer of safflower oil, the microspheres were allowed to sit for 15 minutes, and more oil was then able to be removed.
- In order to completely remove all of the safflower oil and to dry the microspheres, the microspheres were rinsed with hexanes.
- 2mL hexanes were added to each of the micrsophere samples. A cap was placed on the vials and the hexanes were swirled around the vial for 3 minutes.
- After rinsing, the microspheres were vacuum filtered (using Whatman 2 filter paper) and further rinsed with 2mL hexanes. Note: The vacuum filtration and rinsing with hexanes should be performed in a fume hood.
- The microspheres were filtered until completely dry and placed in a new, clean vial.
Note: When preparing microspheres in the future, a label will be added to the vial after the freeze-thaw method. It was noted that the safflower oil removes permanent marker from the glass vials over time.
- Dye will be added to the microspheres the next time. Enough dye will be added for the spheres to have a 1μM concentration. DSC will be run before the addition of dye. The spheres may be subjected to another freeze thaw cycle after the addition of dye to further crosslink the dye into the spheres.