Deparaffinization of tissue by electric field generation and ionization
Paraffin-embedded tissue, which may be disposed on a solid substrate, is prepared by a dry technique that removes paraffin from tissue without adding any liquid to the tissue, thereby rendering the tissue substantially free of paraffin. The dry technique may entail applying heat energy to the tissue effective to melt the paraffin and thereby render it flowable, and applying an electric field. The electric field is effective to impart electrical charge to the paraffin and to move the paraffin out from the tissue due to electrical charge repulsion or attraction, which may be assisted by moving an electrode utilized to generate the electric field relative to the paraffin. The electric field, or both the electric field and the heat energy, may be applied until the tissue is substantially free of paraffin.
1. A method for deparaffinizing a tissue, the method comprising:
providing a tissue embedded with paraffin;
applying heat energy to the tissue effective to melt the paraffin; and
applying an electric field effective to move the paraffin out from the tissue by electrostatic force.
2. The method of claim 1 , comprising
continuing to apply the electric field until the tissue is substantially free of paraffin.
3. The method of claim 1 , wherein the applying heat energy is performed by a process selected from the group consisting of:
placing the tissue sample on a surface and heating the surface;
placing the tissue sample in a chamber and heating the chamber;
applying radiant heat energy;
applying infrared radiation;
applying heat energy so as to heat the paraffin to a temperature in a range from about 35° C. to about 70° C.; and
a combination of two or more of the foregoing.
4. The method of claim 1 , comprising applying the electric field so as to produce a plasma in an ionization region to which the paraffin is exposed.
5. The method of claim 4 , wherein the plasma is produced from air molecules.
6. The method of claim 1 , wherein applying the electric field comprises applying a voltage between a first electrode and a second electrode.
7. The method of claim 6 , wherein applying the electric field produces a corona discharge in a region surrounding the first electrode, and further comprising positioning the first electrode so as to expose the paraffin to the corona discharge.
8. The method of claim 6 , wherein the first electrode has a configuration selected from the group consisting of:
the first electrode comprises a curved feature configured to generate a region of elevated electric field strength surrounding the first electrode; and
the first electrode comprises a curved feature configured to generate a region of elevated electric field strength surrounding the first electrode, wherein the curved feature comprises an edge or a tip of the first electrode.
9. The method of claim 6 , wherein the second electrode is a planar electrode.
10. The method of claim 6 , comprising moving at least one of the first electrode and the tissue relative to the other, in a direction that moves the paraffin away from the tissue.
11. The method of claim 6 , wherein the first electrode comprises an array of electrodes.
12. The method of claim 6 , wherein the tissue is positioned between the first electrode and the second electrode.
13. The method of claim 1 , comprising moving at least one of the electric field and the tissue relative to the other, in a direction that moves the paraffin away from the tissue.
14. The method of claim 1 , comprising applying the heat energy while applying the electric field.
15. The method of claim 1 , wherein providing the tissue comprises a step selected from the group consisting of:
placing the tissue on a solid substrate;
placing the tissue on a slide;
placing the tissue on a plate;
placing the tissue on beads;
placing the tissue on a porous medium;
placing the tissue on a filter;
placing the tissue in a liquid container; and
placing the tissue on a substrate surface having a higher surface energy than the paraffin.
16. The method of claim 1 , wherein providing the tissue comprises positioning the tissue in or above a container, and further comprising collecting the paraffin in the container after moving the paraffin out from the tissue.
17. The method of claim 1 , comprising staining the tissue after removing the paraffin.
18. The method of claim 17 , wherein staining the tissue comprises staining with an immunohistochemical staining agent.
19. The method of claim 1 , comprising isolating nucleic acids from the tissue after removing the paraffin.