Biological laser printing for tissue microdissection via indirect photon-biomaterial interactions

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 characterized by cellular heterogeneity coated on top of the interlayer opposite to the support;

wherein the interlayer comprises a metal oxide;

providing a source of photon energy;

selecting particular cells, groups of cells, or sub-cellular regions to be microdissected from the tissue by the 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 the particular cells, groups of cells, or sub-cellular regions across a gap between the target substrate and the receiving substrate or collection vessel and onto the receiving substrate or into the collection vessel.

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 titanium dioxide.

11. The method of claim 1 , wherein the interlayer is about 1 Angstroms to about 10 microns thick.

12. The method of claim 1 , wherein the photon energy source is a pulsed laser.

13. The method of claim 12 , wherein the laser energy has a fluence between about 1 and about 1000 mJ/cm 2 .

14. The method of claim 12 , wherein the laser energy has a fluence of at least about 0.1 mJ/cm 2 .

15. The method of claim 12 , wherein the laser energy has a fluence of at least about 1 nJ/cm 2 .

16. The method of claim 1 , wherein the photon energy source is a continuous laser.

17. The method of claim 1 , wherein the photon energy source is a flash lamp.

18. The method of claim 1 , wherein the photon energy source is a maser.

19. The method of claim 1 , wherein the target substrate and the photon energy source are moveable with respect to each other.

20. 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.

21. The method of claim 1 , wherein the target substrate comprises a plurality of regions comprising different cells and regions of tissues.

22. 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.

23. The method of claim 22 , wherein the dehydrating step comprises successive washes in alcohol/water solutions having increasing percentages of alcohol.

24. The method of claim 23 , further comprising a final wash in about 100% alcohol.

25. The method of claim 22 , wherein the thin section contains paraffin.

26. The method of claim 25 , further comprising the step of:

deparaffinizing the thin section.

27. The method of claim 22 , further comprising the step of:

staining the thin section.

28. The method of claim 27 , 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.

29. The method of claim 1 , wherein the receiving substrate or collection vessel comprises a microcentrifuge tube.

30. The method of claim 29 , further comprising the step of:

performing an assay, biochemical or physical, on the collected cells, groups of cells, or sub-cellular regions.

31. The method of claim 1 , wherein the particular cells, groups of cells, or sub-cellular regions comprise a substantially homogenous population of cells.

32. The method of claim 1 , wherein the particular cells, groups of cells, or sub-cellular regions comprise cells of a specific and defined population of cells.

33. The method of claim 1 , wherein the particular cells, groups of cells, or sub-cellular regions comprise diseased cells.

34. The method of claim 1 , wherein the particular cells, groups of cells, or sub-cellular regions are selected by visual identification using a microscope objective, a video camera, and a video screen.

35. The method of claim 1 , wherein the histopathologically processed or frozen tissue/cell materials are histopathologically processed or frozen tissue.

36. The method of claim 1 , further comprising:

performing the method a plurality of times;

wherein the plurality of transferred particular cells, groups of cells, or sub-cellular regions are collected in the same collection vessel; and

performing a physical assay, biochemical assay, visual assay, biological assay, proteomic assay, genomic assay, or general molecular biology assay on the collected cells, groups of cells, or sub-cellular regions.

37. The method of claim 1 , wherein the receiving substrate or collection vessel is a collection vessel.