Semiconductor nanocrystal-based phagokinetic tracking
Methods for determining metabolic properties of living cells through the uptake of semiconductor nanocrystals by cells. Generally the methods require a layer of neutral or hydrophilic semiconductor nanocrystals and a layer of cells seeded onto a culture surface and changes in the layer of semiconductor nanocrystals are detected. The observed changes made to the layer of semiconductor nanocrystals can be correlated to such metabolic properties as metastatic potential, cell motility or migration.
1. A method for phagokinetic tracking, comprising:
providing a culture surface for supporting biological activity of living cells;
depositing semiconductor nanocrystals on the culture surface, thereby resulting in deposited semiconductor nanocrystals;
noting the starting positions of the deposited semiconductor nanocrystals on the culture surface;
seeding the cells onto the culture surface;
allowing the seeded cells to move and take up the deposited semiconductor nanocrystals; and
detecting a phagokinetic track of the cells by observing changes in the positions of the deposited semiconductor nanocrystals after sufficient time has elapsed to allow the seeded cells to move and take up the deposited semiconductor nanocrystals, wherein the changes in the positions of the deposited semiconductor nanocrystals indicate the phagokinetic track of the cell and wherein the detection of a phagokinetic track of the cells indicates the cells having the properties of motility, migration, movement and/or metastatic potential.
2. The method of claim 1 further comprising adding a marker to be taken up by the cells, wherein the marker is selected from the group consisting of other semiconductor nanocrystals that are different from the deposited semiconductor nanocrystals and an organic fluorescent dye.
3. The method of claim 2 wherein the organic fluorescent dye is selected from the group consisting of actinomycin D, acridine orange, bisbenzimide, 4′,6-Diamidino-2-phenylindole (DAPI), propidium iodide, ethidium bromide, carbocyanines, pyridinium dibromides, tetramethylrhodamine ethyl ester (TMRE), ceramides, fluorescein-labeled phalloidin, GFP, DsRed, YCP, CFP and BFP.
4. The method of claim 1 wherein the detecting-takes place over a time period between ten minutes and 10 days.
5. The method of claim 1 wherein each of the semiconductor nanocrystals comprises a core semiconductor covered by a hydrophilic outermost shell.
6. The method of claim 5 wherein the core is CdSe.
7. The method of claim 6 wherein the outermost shell is a siloxane.
8. The method of claim 7 wherein each of the semiconductor nanocrystals further comprises an intermediate shell of ZnS.
9. The method of claim 5 wherein the outermost shell comprises a stabilizing group which carries a positive charge.
10. The method of claim 1 further comprising adding other semiconductor nanocrystals that have a different size from the deposited semiconductor nanocrystals.
11. The method of claim 1 wherein the depositing comprises depositing the semiconductor nanocrystals in patterned arrangements onto the culture surface.
12. The method of claim 1 wherein the depositing comprises depositing the semiconductor nanocrystals in linear arrangements onto the culture surface.
13. The method of claim 1 wherein the depositing comprises depositing the semiconductor nanocrystals homogenously onto the culture surface.
14. The method of claim 1 wherein the culture surface comprises an extracellular matrix.
15. The method of claim 14 wherein the extracellular matrix comprises proteins.
16. The method of claim 15 wherein the proteins are selected from the group consisting of collagen, laminin, fibronectin, elastin, nidogen, enactin, and proteoglycan.
17. The method of claim 1 wherein the cells are cancerous.
18. The method of claim 1 wherein the cell properties are correlated with metastatic potential.
19. The method of claim 1 further comprising measuring or quantifying cell motility.
20. The method of claim 1 further comprising measuring or quantifying cell chemotactic behavior.
21. The method of claim 7 wherein each of the semiconductor nanocrystals further comprises an intermediate shell of CdS.
22. The method of claim 5 wherein the outermost shell comprises a stabilizing group which carries a negative charge.