Nanoshell Method for Nanoparticle Attachment Protocol
Protocol for attaching 5nm gold nanoparticles to yeast cells:
1. Measured 0D600 of cells = 10.4
2. Transferred 2mL of cells to a 2mL Eppendorf tube and centrifuge at 5000rpm for 20 minutes to collect cells.
3. Decant supernatant, resuspend in 1mL of PBS
4. Centrifuge at 5000rpm for 20 minutes
5. Decant supernatant, resuspend in 1mL of PBS
6. Set up 5 tubes according to the following table:
| % NP |
0% |
20% |
40% |
60% |
80% |
| Vol yeast |
100 µL |
100 µL |
100 µL |
100 µL |
100 µL |
| Vol NP |
0 |
400 µL |
800 µL |
1.2 mL |
1.6 mL |
| Vol PBS |
1.9 mL |
1.5 mL |
1.1 mL |
700 µL |
300 µL |
*Vol yeast corresponds to the volume of cell solution taken from the tube containing PBS/yeast (Step 5)
7. Incubate at room temperature for half an hour
8. Centrifuge at 4000rpm for 15 minutes
9. Resuspend in 1mL of 1X TE buffer
Source: Jiang, N., Yang, X-Y., Ying, G-L., Shen, L., Liu, J., Geng, W., Dai, L-J., Liu, S-Y., Cao, J., Tian, G., Sun, T-L., Li, S-P., and Su, B-L. (2015) “Self-repairing” nanoshell for cell protection. Chem Sci., 6:486-491. Retrieved from http://pubs.rsc.org/en/Content/ArticleLanding/2015/SC/c4sc02638a#!divAbstract
