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 ...view middle of the document...
I will also be looking
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 solvent, I will be able to see whether actin has decreased, which means that it could have attached to QDs. The sediments will be boiled and the upper solvents of the boiled material will be used in Western Blotting to see if there is actually actin in the sediment, which will further prove that actin and QDs are able to attach.