Research Proposal

The Harmful Influence of Quantum Dots on Cytoskeleton Meshwork

Recent study suggests that Quantum Dot exposure causes significant morphological changes to macrophages; cells became round and condensed with fewer surface protrusions in response to QD treatment, it has also been demonstrated that QD’s intracellular localization undermined the capability of macrophagic phagocytosis.

Since actin is fundamental to cellular morphological changes, it would be worthwhile to investigate the activity between QDs and the filamentous actin. In order for the QDs to affect the microfilament meshwork, QDs might be able to attach itself to actin. Through confocal laser scanning microscopy and immunoprecipitaion I hope to prove that QDs are able to attach with actin.

Cell Culture
I plan to culture three types of cells, 293, HepG2, Hela. A viability test will be done to establish a concentration for further experiment that does not exert harm on cell growth and survival of all three types of cells. I will also be looking

Confocal Microscopy
The nuclei will be stained with 4',6-diamidino-2-phenylindole (DAPI, blue) and the cytoskeleton with FITC-conjugated phalloidin (green). The fluorescence for nuclei, cytoskeleton and QDs will be assessed through confocal laser scanning microscopy. All three types of cells will be examined by confocal microscopy. Through this experiment, I will be observing the relative position of QDs in the cell, and the reduction in the density of actin meshwork and in the numbers of actin rich surface protrusion in cells upon exposure to QDs.

Protein Pulldown and Western Blot
I will first expose the cells with QDs for 24 hours, and then extract the cell protein. After extraction, most of the proteins will be in the solvent, and can be separated from QDs, cell membranes, DNA etc. by centrifugation. By Western Blotting the