Gold implantation/deposition of biological samples for laser desorption three dimensional depth profiling of tissues

• US 20050035284A1
• Filed: 06/04/2004
• Published: 02/17/2005
• Est. Priority Date: 06/06/2003
• Status: Active Grant

• Current Assignee: Ionwerks, Inc.
• Original Assignee: Ionwerks, Inc.
• Inventors: Schultz, J. Albert, Egan, Thomas F., Tempez, Agnes, Ugarov, Michael V., Della-Negra, Serge, Le Bayec, Yvon
• Granted Patent: US 6,989,528 B2
• Time in Patent Office: Days
• US Class Current: 250/287
• CPC Class Codes: H01J 49/0031 Step by step routines describing the use of the apparatus H01J49/0081 takes precedence
• CPC Class Codes: H01J 49/0418 for laser desorption, e.g. matrix-assisted laser desorption/ionisation [MALDI] plates or surface enhanced laser desorption/ionisation [SELDI] plates
• CPC Class Codes: H01J 49/164 Laser desorption/ionisation, e.g. matrix-assisted laser desorption/ionisation [MALDI] sample holders H01J49/0418

Abstract

The present invention enhances the laser desorption of biological molecular ions from surfaces by creating a surface localized MALDI particle matrix by ion implantation of low energy ionized clusters (gold, aluminum, etc.) or chemically derivatized clusters into the near surface region of the sample. MALDI analysis of the intact biomolecules on the surface or within a narrow subsurface region defined by the implantation range of the ions can then be performed by laser desorption into a mass spectrometer or, in a preferred embodiment, into a combined ion mobility orthogonal time of flight mass spectrometer.

Claims

1. An analytical instrument for the characterization and analysis of a sample comprising:

a MALDI sampling device comprising a sample stage, said sample stage capable of accommodating a sample;

a component selected from the group consisting of a metal ion cluster beam source, an inorganic cluster ion beam source, a vapor deposition system, a laser ablation system, a desorption source, and any combination thereof, said component being capable of adding a matrix to said sample, said component being fluidly coupled to said MALDI sampling device;

a laser coupled to said MALDI sampling device, said laser being capable of desorbing material from said sample;

an ion mobility cell having a drift tube, said mobility cell coupled to said MALDI sampling device and capable of receiving sample from said MALDI sampling device; and

,

a time-of-flight mass spectrometer having a flight tube positioned orthogonally to said drift tube, said flight tube fluidly coupled to said drift tube.

2. The instrument of claim 1, wherein the metal ion cluster beam is a gold ion cluster beam.

3. The instrument of claim 2, wherein the gold cluster ion beam delivers gold clusters in the range Au₁₀₀-Au₃₀₀ and having energy within the range of a few hundred eV/gold atom, to an energy of several hundreds of keV/gold atom.

4. The instrument of claim 2, wherein the gold cluster beam has a spatial resolution of less than one micron.

5. The instrument of claim 1, wherein the MALDI sampling device is an atmospheric MALDI device wherein the MALDI ions are desorbed at atmospheric pressure and transported through a differential pumping interface into the mass spectrometer.

6. The instrument of claim 1, further comprising a differentially pumped interface between the MALDI sampling device at atmospheric pressure and the mass spectrometer, said differentially pumped interface is an ion mobility cell operating at a pressure of from about 1-10 Torr up to atmospheric pressure.

7. The instrument of claim 1, wherein the drift tube has a carrier gas comprising nitrogen or helium at 2 Torr pressure.

8. The instrument of claim 1, further comprising a data acquisition electronics and software system.

9. The instrument of claim 1, wherein the sample stage is an X-Y movable stage.

10. The instrument of claim 1, wherein the sample stage is housed in a low pressure chamber.

11. The instrument of claim 1, wherein the component is a vapor deposition system.

12. The instrument of claim 11, wherein the sample stage is a rotatable sample stage.

13. The instrument of claim 1, wherein the component is a laser ablation deposition system.

14. The instrument of claim 13, wherein the sample stage is a rotatable sample stage.

15. The instrument of claim 1, wherein the sample stage is a desorption source coupled to an ion mobility cell.

16. The instrument of claim 15, wherein the deposition source comprises a laser ablation source, an electrospray source or a combination thereof.

17. The instrument of claim 15, wherein the instrument further comprises gating electronics for size selecting the mobility ion.

18. The instrument of claim 15, wherein the sample stage is cryogenically cooled.

19. A method for the collection of mass spectrometric data from a sample, comprising the steps of:

adding matrix to the sample with a component selected from the group consisting of a metal ion cluster beam, an inorganic cluster ion beam, a vapor deposition system, a laser ablation deposition system, a desorption source, and any combination thereof;

laser desorbing chemical species from said sample;

separating the desorbed chemical species in a drift tube by ion mobility; and

,

further separating the chemical species in a time-of-flight mass spectrometer.

20. The method of claim 19, wherein the step of adding matrix to the sample comprises adding matrix with a metal ion cluster beam.

21. The method of claim 20, wherein said step of adding matrix with a metal ion cluster beam comprises microfocusing said metal ion cluster beam onto a spot on said sample.

22. The method of claim 19, further comprising the step of microdissecting said sample.

23. The method of claim 19, wherein the metal ion cluster beam is a gold ion cluster beam.

24. The method of claim 19, wherein the step of laser desorbing comprises laser desorbing in an atmospheric MALDI device.

25. The method of claim 19, wherein the step of separating the desorbed chemical species in a drift tube by ion mobility comprises separating in a nitrogen or helium mobility carrier at about 1 Torr pressure.

26. The method of claim 19, further comprising the step of acquisition of two dimensional mass-volume data.

27. The method of claim 19, further comprising the step of moving the sample in either or both of the X and Y directions.

28. The method of claim 19, wherein the step of adding matrix to the sample comprises adding matrix to the sample with vapor deposition.

29. The method of sentence 28, further comprising the step of rotating the sample.

30. The method of sentence 20, wherein the step of adding matrix to the sample comprises adding matrix to the sample with a laser ablation deposition system.

31. The method of sentence 30, further comprising the step of rotating the sample.

32. The method of sentence 20, wherein the step of adding matrix to the sample comprises adding matrix to the sample with a desorption source coupled to a mobility cell.

33. The method of sentence 32 wherein the desorption source comprises a laser ablation source, an electrospray ionization source, or a combination thereof.