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1. WO2016112171 - TRANSFERT D'ÉCHANTILLON ASSISTÉ PAR LASER À POINTE AMÉLIORÉE, POUR SPECTROMÉTRIE DE MASSE DE BIOMOLÉCULE

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

[ EN ]

CLAIMS

We claim at least the following:

1. A method for transferring molecules from a film to a collection device for analysis via an atomic force microscope and a pulsed laser comprising:

positioning a gold-coated silicon tip of the atomic force microscope above the film to image a sample of the molecules, the gold-coated silicon tip having a radius of curvature and being positioned at a first distance from the film;

focusing the pulsed laser onto the gold-coated silicon tip of the atomic force microscope;

ablating the sample from a plurality of ablation spots on the film onto the collection device, the collection device being suspended at a first position relative to the gold-coated silicon tip, wherein a first amount of material from the sample is ablated from each of the ablation spots and transferred onto the collection device.

2. The method of claim 1, wherein the collection device comprises a metal wire.

3. The method of claim 1 or claim 2, wherein the molecules comprise at least one of: anthracene or rhodamine 6G, and wherein the metal wire is cut and affixed to a metal target using double-sided conductive tape, and wherein the molecules are analyzed by laser desorption ionization (LDI) using a commercial laser desorption time-of-flight mass spectrometer.

4. The method of claim 1 or claim 2, wherein the molecules comprise at least one of: angiotensin II or bovine insulin, and wherein the molecules are dissolved in a saturated matrix solution and deposited on a target for matrix-assisted laser desorption ionization (MALDI) analysis.

5. The method of any one of claims 1-4, wherein the radius of curvature of the gold-coated silicon tip is about 30 nm and the first distance is about 10 nm.

6. The method of any one of claims 1-5, wherein the first position relative to the gold-coated silicon tip is located about 300 μηι vertically from the gold-coated silicon tip and about 100 μηι horizontally from the gold-coated silicon tip.

7. The method of any one of claims 1-6, wherein a polarization of a laser beam of the pulsed laser is adjusted via a half wave plate, the polarization of the laser beam being perpendicular at 355 nm or parallel at 532 nm.

8. The method of claim 7, wherein the laser beam is focused with a 25 cm calcium fluoride (CaF2) lens to irradiate the gold-coated silicon tip at an angle 83° that is from normal.

9. The method of any one of claims 1-8, further comprising engaging the gold-coated silicon tip in a tapping mode with the sample at an amplitude set point of 1 V.

10. The method of any one of claims 1-9, wherein the sample is ablated via about 30 laser shots at more than about 3 seconds per ablation spot.

11. The method of any one of claims 1-10, wherein the analysis is mass spectrometry.

12. The method of claim 11, wherein the resonant frequency is about 300 kHz.

13. A system for capturing and transferring molecules from a surface for mass spectrometry comprising:

a pulsed laser configured to:

focus onto a gold-coated silicon tip of an atomic force microscope; and ablate the molecules from a plurality of ablation spots on the surface; and

a collection device configured to:

capture the molecules ablated from the plurality of ablation spots, the collection device being suspended at a first position relative to the gold-coated silicon tip of the atomic force microscope.

14. The system of claim 13, wherein the pulsed laser is a 355 nm or a 532 nm pulsed laser.

15. The system of claim 13 or claim 14, wherein the pulsed laser is focused onto the gold-coated silicon tip at an angle of about 45° in a horizontal plane relative to the surface and at an angle of about 8° in a vertical plane relative to the surface.

16. The system of any one of claims 13-15, wherein the collection device comprises a capture wire.

17. The system of claim 16, wherein the capture wire is a silver capture wire and a tip of the silver capture wire is flattened to produce a ribbon of about 400-500 um in width.

18. The system of any one of claims 13-17, wherein the first position relative to the gold-coated silicon tip is located about 300 μηι above the gold-coated silicon tip and about 100 um horizontally from the gold-plated silicon tip.

19. The system of any one of claims 13-18, wherein a focal point of a laser beam of the pulsed laser is about 4 cm beyond the gold-coated silicon tip and a resulting diameter where the laser beam intersects the gold-coated silicon tip is about 600 um.

20. The system of claim 19, wherein a distance between the collection device and at least one of the plurality of ablation spots is about 100 μηι.