Subject Number : S-16-NM-0056
Support Type : 機器利用
Proposal Title (English) : Nanotoxicity　（Usage of cell analyzer and confocal microscope）
Username (English) : Qingqing Sun
Affiliation (English) : Graduate School of Advanced Science and Engineering, Waseda University

1. Summary
In the year of 2016, I used the cell analyzer and confocal microscope to check the cellular uptake and localization of nanoparticles. These machines are very good tools with convenient operation to us.

2. Experimental
2.1 Cell analyzer
After incubation with nanoparticles for certain time, cells were collected and stained with suitable dyes. Following that, cell analyzer was used to check the cellular uptake. Basically, suitable detection parameters were fixed firstly, and then cells were filtered and detected.
2.2 Conforcal laser scanning microscope
Cells were cleaned, fixed and stained with suitable dyes after exposure with nanoparticles for certain time. Finally, confocal microscope machine was used to observe cells and nanoparticles at the 63X objective.

3. Results and Discussion
For the cellular uptake, our work found that larger nanoparticles owned a higher cellular uptake, which confirmed to the confocal microscope observation results.

Figure 1. Change in uptake by NCI-H292 cells with increasing nanoparticle suspensions’ concentration and size. (A) Histograms of cellular uptake. (B) Curves showing the change in cellular uptake with increasing size and concentration of nanoparticle suspensions. (C) Confocal microscope pictures of the cellular uptake of nanoparticles. The blue ones show the nuclei of cells stained by DAPI; the green ones show the FITC-TiO2-PEG nanoparticles as directed by the white arrows.

4. Others
Acknowledgements:
In here, we thank Xianglan Li, Hattori shinya, Min-hua chen and Morita hiromi for training and trouble shooting and discussion of machine operation and work troubles.

5. Publication/Presentation
(1) Sun, Qingqing, et al. “Uniform TiO2 nanoparticles induce apoptosis in epithelial cell lines in a size-dependent manner.” Biomaterials Science (2017).
(2) Qingqing Sun, poster, “Uniform TiO2 nanoparticles induce apoptosis in epithelial cell lines in a size-dependent manner” in the japan biomaterials society, 2016/11/23.
(3) Qingqing Sun, oral, “Uniform TiO2 nanoparticles induce apoptosis in epithelial cell lines in a size-dependent manner” in the 第26回インテリジェント材料・システムシンポジウム　プログラム, 2017/01/11.

6. Patent
N/A