Claims
- 1. A method for microdissecting histopathologically processed tissue/cell materials comprising the steps of:
providing a receiving substrate or collection vessel; providing a target substrate comprising a photon-transparent support, a photon absorbent interlayer coated on the support, and a transfer material comprising histopathologically processed or frozen tissue/cell materials coated on top of the interlayer opposite to the support; providing a source of photon energy; and directing the photon energy through the transparent support so that it strikes the interlayer;
wherein a portion of the interlayer is energized by absorption of the photon energy; and wherein the energized interlayer causes a transfer of a portion of the histopathologically processed tissue/cell material across a gap between the target substrate and the receiving substrate and onto the receiving substrate or into the collection vesicle.
- 2. The method of claim 1, wherein the energized interlayer remains substantially intact and adhered to the substrate.
- 3. The method of claim 1, wherein the energized interlayer is removed.
- 4. The method of claim 1,
wherein the transfer material comprises a biomatrix; and wherein the transfer occurs through evaporation of a portion of the biomatrix adjacent to the energized interlayer.
- 5. The method of claim 1, wherein the transfer occurs through photomechanical shock.
- 6. The method of claim 1, wherein the transfer occurs through photothermal shock.
- 7. The method of claim 1, wherein the transfer occurs through ablation of the interlayer.
- 8. The method of claim 1, wherein the photon transparent support comprises quartz.
- 9. The method of claim 1, wherein the photon transparent support comprises a material selected from the group consisting of a glass, a salt, and a polymer.
- 10. The method of claim 1, wherein the interlayer comprises a metal or a metal oxide.
- 11. The method of claim 1, wherein the interlayer comprises a material selected from the group consisting of titanium metal, titanium dioxide, chrome, molybdenum, gold, polymer, and combinations thereof.
- 12. The method of claim 1, wherein the interlayer is about 1 Angstroms to about 10 microns thick.
- 13. The method of claim 1, wherein the photon energy source is a pulsed laser.
- 14. The method of claim 13, wherein the laser energy has a fluence between about 1 and about 1000 mJ/cm2.
- 15. The method of claim 13, wherein the laser energy has a fluence of at least about 0.1 mJ/cm2.
- 16. The method of claim 13, wherein the laser energy has a fluence of at least about 1 nJ/cm2.
- 17. The method of claim 1, wherein the photon energy source is a continuous laser.
- 18. The method of claim 1, wherein the photon energy source is a flash lamp.
- 19. The method of claim 1, wherein the photon energy source is a maser.
- 20. The method of claim 1, wherein the target substrate, the receiving substrate, and the photon energy source are moveable with respect to each other.
- 21. The method of claim 1, wherein the step of providing a target substrate is repeated one or more times using one or more additional target substrates comprising one or more different transfer materials.
- 22. The method of claim 1, wherein the target substrate comprises a plurality of regions comprising different cells and regions of tissues.
- 23. The method of claim 1, wherein the biological material comprises histopathologically processed tissue/cells processed according to the steps of:
providing a thin section of the tissue; placing the thin section onto the interlayer; dehydrating the thin section with an alcohol; and rehydrating the thin section with a buffer containing glycerol.
- 24. The method of claim 23, wherein the dehydrating step comprises successive washes in alcohol/water solutions having increasing percentages of alcohol.
- 25. The method of claim 24, further comprising a final wash in about 100% alcohol.
- 26. The method of claim 23, wherein the thin section contains paraffin.
- 27. The method of claim 26, further comprising the step of:
deparaffinizing the thin section.
- 28. The method of claim 23, further comprising the step of:
staining the thin section.
- 29. The method of claim 28, wherein the staining is performed using a stain selected from the group consisting of but not limited to eosin, hematoxylin, methyl green, and Wright's stain.
- 30. The method of claim 1, wherein the biological material comprises histopathologically processed or frozen tissue and further comprising the step of:
selecting particular cells, groups of cells, or sub-cellular regions to be microdissected from the tissue by the photon energy.
- 31. The method of claim 30, wherein the receiving substrate comprises a receptacle for collecting the transferred cells, groups of cells, or sub-cellular regions.
- 32. The method of claim 31, further comprising the step of:
performing an assay, biochemical or physical, on the collected cells, groups of cells, or sub-cellular regions.
Parent Case Info
[0001] This application claims the benefit of U.S. Provisional Patent Application No. 60/476,377, filed on Jun. 6, 2003 and U.S. Provisional Patent Application No. 60/542,841, filed on 02/10/2004. All applications named are incorporated by reference. The U.S. Patent Application to Ringeisen et al., titled “Biological Laser Printing via Indirect Photon-Biomaterial Interactions,” designated NC 84,621, filed on the same day as the present application is also incorporated by reference.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60476377 |
Jun 2003 |
US |
|
60542841 |
Feb 2004 |
US |