Devices and processes for analyzing nucleic acid damage and repair using electrophoresis
Systems, methods, and devices are provided for assessing DNA damage and repair in cells by measuring DNA migration under electrophoresis. In one exemplary embodiment, a microarray configured to hold cells in a predetermined spatial relationship is employed to improve accuracy, speed, and reliability of such measurements. In another embodiment, a self-contained cassette having a matrix material disposed therein can be used to create a substantially uniform environment for analyzing DNA damage and repair. Fluid can be circulated through the cell to assist in creating spatial patterns on the matrix material, or alternatively, the matrix material can already include a microarray pattern disposed thereon. Various methods and systems that take advantage of such microarrays and cassettes are also provided.
1. A method for analyzing a nucleic acid, comprising:
disposing at least one cell having DNA to be analyzed into each of a plurality of spatially-defined cell capture locations formed in a solidified matrix material;
positioning an overlay over the matrix material;
utilizing the overlay to apply at least one reagent to at least one cell capture location of the plurality of spatially defined cell capture locations or a portion of the matrix material; and
analyzing at least one cell following application of the reagent for nucleic acid fragmentation.
2. The method of claim 1 , further comprising:
removing the overlay from the matrix material;
performing electrophoresis on the plurality of cells; and
analyzing nucleic acid fragmentation based on comet parameters resulting from nucleic acid migration.
3. The method of claim 1 wherein the step of utilizing the overlay further comprises utilizing an overlay having at least one reagent disposed thereon to apply the reagent by contact with the cells.
4. The method of claim 1 wherein the step of utilizing the overlay further comprises utilizing an overlay having at least one opening to allow at least one reagent to be passed therethrough.
5. The method of claim 1 , wherein the matrix material is an agarose gel.
6. The method of claim 1 , wherein a thickness of the solidified matrix material is in the range of about 10 microns to about 300 microns.
7. The method of claim 1 , wherein a depth of the plurality of spatially-defined cell capture locations is in the range of about 4 micrometers to about 20 micrometers.
8. The method of claim 1 , wherein the step of disposing cells in the spatially-defined cell capture locations further comprises applying a fluid comprising the cells onto the surface of the matrix material.
9. The method of claim 1 , wherein the step of disposing cells in the spatially-defined cell capture locations further comprises printing cells onto the matrix material.
10. The method of claim 1 , wherein the step of applying at least one reagent to the cells further comprises applying multiple, different reagents to the cells.
11. The method of claim 1 , further comprising moving the overlay with respect to the matrix material while applying at least one reagent to the cells.
12. The method of claim 1 , further comprising:
applying a second matrix material to the first matrix material after cells have been applied over the surface of the first matrix material; and
allowing the second matrix material to solidify to approximately maintain a location of the cells with respect to the first matrix material.
13. The method of claim 1 , further comprising:
placing a divider over the matrix material; and
adding a second matrix material having a plurality of cells disposed therein above the divider.
14. The method of claim 1 , further comprising:
applying a matrix material to a substrate, the matrix material being the matrix material in which a plurality of spatially-defined cell capture locations that receive at least one cell having DNA to be analyzed are formed;
contacting the matrix material with a template having a pattern of protuberances to form the plurality of spatially-defined cell capture locations in the matrix material; and
removing the template from the matrix material.
15. A method for manufacturing a nucleic acid analysis device, comprising:
applying a matrix material to a substrate, the matrix material including an agarose gel;
contacting the matrix material with a template having a pattern of protuberances to form spatially-defined cell capture locations in the matrix material;
removing the template from the matrix material; and
allowing the matrix material to solidify such that a plurality of spatially-defined cell capture locations are formed therein at locations based on the pattern of protuberances, each cell capture location of which is configured to receive one or more cells having DNA to be analyzed.
16. The method of claim 15 , wherein a thickness of the solidified matrix material is in the range of about 10 microns to about 300 microns.
17. The method of claim 15 , wherein a depth of the plurality of spatially-defined cell capture locations is in the range of about 4 micrometers to about 20 micrometers.