Apparatus and method for sub-micrometer elemental image analysis by mass spectrometry

• US 9,312,111 B2
• Filed: 03/30/2015
• Issued: 04/12/2016
• Est. Priority Date: 04/02/2014
• Status: Active Grant

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Bendall, Sean C., Angelo, Robert M., Nolan, Garry P.
• Primary Examiner(s): Vanore, David A
• Application Number:
  US14/673,279
• Publication Number:
  US 20150287578A1
• Time in Patent Office: 379 Days
• Field of Search: 250/282, 250/287, 250/288, 250/281, 250/307, 250/309, 250/423.R, 250/305, 250/306, 250/397, 250/398, 250/492.3
• US Class Current: 1/1
• CPC Class Codes: H01J 49/0004 Imaging particle spectrometry
• CPC Class Codes: H01J 49/0036 Step by step routines describing the handling of the data generated during a measurement
• CPC Class Codes: H01J 49/0409 Sample holders or containers containers for retaining a material to be analyzed, B01L3/50, for DNA, C12Q1/6834, for biological materials, G01N33/543
• CPC Class Codes: H01J 49/0413 for automated handling
• CPC Class Codes: H01J 49/142 using a solid target which is not previously vapourised
• CPC Class Codes: H01J 49/40 Time-of-flight spectrometers H01J49/36 takes precedence
• CPC Class Codes: H01J 49/401 characterised by orthogonal acceleration, e.g. focusing or selecting the ions, pusher electrode

Abstract

A mass spectrometer system having: a primary ion source capable of irradiating a segment on a planar sample with a beam of primary ions, an orthogonal ion mass-to-charge ratio, the analyzer being configured to separate secondary elemental atomic ions according to their mass-to-charge ratio by time of flight; an ion detector for detecting secondary elemental atomic ions and producing mass spectra measurements; and a synchronizer. In the system, the beam of primary ions scans across the planar sample in two dimensions and the synchronizer associates the mass spectra measurements with positions on the planar sample.

Claims

1. A mass spectrometer system for elemental analysis of a planar sample, the mass spectrometer system comprising:

a) a sample interface comprising a holder that is configured to hold a substrate comprising a planar sample;

b) a primary ion source capable of irradiating a segment on the planar sample with a beam of primary ions that is less than 1 mm in diameter, wherein irradiation of the planar sample with the primary ions results in the production of secondary elemental atomic ions derived from staining elements associated with the planar sample;

c) an orthogonal ion mass-to-charge ratio analyzer positioned downstream of sample interface, the analyzer being configured to separate secondary elemental atomic ions according to their mass-to-charge ratio by time of flight;

d) a main ion detector for detecting the secondary elemental atomic ions and producing mass spectra measurements; and

e) a synchronizer that associates the mass spectra measurements with positions on the planar sample,

wherein the system is configured so that so that the beam of primary ions scans across the planar sample in two dimensions.

2. The mass spectrometer system of claim 1, wherein the system is configured so that the primary ion source continuously irradiates the planar sample as the beam of primary ions scans across the planar sample.

3. The mass spectrometer system of claim 1, wherein the diameter of the beam of primary ions is tunable to a selected diameter in the range of 10 nm to □

mm.

4. The mass spectrometer system of claim 1, wherein the system is configured to:

perform a first scan a first area of said planar sample to collect a first set of data; and

perform a second scan said first area of said planar sample to collect a second set of data;

wherein the diameter of the beam of primary ions of the first scan is at least 2×

larger than the diameter of the beam of primary ions of the second scan, and the synchronizer associates the mass spectra measurements with a position on the planar sample and the timing of the scans.

5. The mass spectrometer system of claim 4, wherein the system is configured to

perform a third scan the first area of said planar sample to collect a third set of data, using a beam of primary ions that has a diameter that is smaller than the diameter of the beam of the second scan.

6. The mass spectrometer system of claim 4, wherein the first set of data is collected using a beam of primary ions that has a diameter in the range of 100 nm to 100 μ

m and the second set of data is collected using a beam of primary ions that has a diameter in the range of 10 nm to 1 μ

m.

7. The mass spectrometer system of claim 1, wherein the system is configured to:

a) move the planar sample to a defined position, thereby presenting a first area on planar sample to the beam of primary ions and

b) raster the beam of primary across said first area to produce a plurality of mass spectra measurements for said first area.

8. The mass spectrometer system of claim 7, wherein the system is configured to:

c) move the planar sample to a second defined position after said plurality of mass spectra measurements for said first area have been collected, thereby presenting a second area on the planar sample to the beam of primary ions; and

d) raster the beam of primary across said second area to produce a plurality of mass spectra measurements for said second area.

9. The spectrometer system of claim 8, further comprising repeating steps c) and d) until sufficient data has been collected.

10. The mass spectrometer system of claim 1, wherein the planar sample is mounted on a conductive substrate where the substrate surface has been depleted of atoms that give rise to said secondary elemental atomic ions.

11. The mass spectrometer system of claim 1, wherein said beam of primary ions ionize mass tags in the planar sample, and said mass spectra measurements comprise the abundance and identify of said mass tags.

12. The mass spectrometer system of claim 1, wherein said beam of primary ions is an oxygen, xenon, argon, gold, bismuth gallium, SF₆ or C₆₀ ion beam.

13. The mass spectrometer system of claim 1, wherein said mass tags comprise elements having an atomic number of 21-29, 39-47, 57-79 or 89.

14. The mass spectrometer system of claim 1, wherein the system comprises an energy filtering means configured to enrich for the secondary elemental atomic ions before they are separated by the analyzer.

15. A method for reconstructing an image of a planar sample, comprising:

(a) placing said planar sample comprising staining elements into the holder of the mass spectrometer system comprising;

i. a sample interface comprising a holder that is configured to hold a substrate comprising a planar sample;

ii. a primary ion source capable of irradiating a segment on the planar sample with a beam of primary ions that is less than 1 mm in diameter, wherein irradiation of the planar sample with the primary ions results in the production of secondary elemental atomic ions derived from staining elements associated with the planar sample;

iii. an orthogonal ion mass-to-charge ratio analyzer positioned downstream of sample interface, the analyzer being configured to separate secondary elemental atomic ions according to their mass-to-charge ratio by time of flight;

iv. a main ion detector for detecting the secondary elemental atomic ions and producing mass spectra measurements; and

v. a synchronizer that associates the mass spectra measurements with positions on the planar sample, wherein the system is configured so that so that the beam of primary ions scans across the planar sample in two dimensions;

(b) producing a data file containing mass spectra measurements for an area of the planar sample using the mass spectrometer system, wherein said mass spectra measurements are associated with positions on the planar sample; and

(c) reconstructing an image of said of the planar sample using the mass spectra measurements.

16. The method of claim 15, further comprising sending said data file and/or the image to a remote location.

17. The method of claim 15, wherein the method comprises:

performing a survey scan of the planar sample to identify regions of interest; and

re-scanning the regions of interest;

i) by rastering said ion beam at a higher resolution than the survey scan;

ii) using a beam of primary ions having a smaller diameter than the survey scan;

iii) with a longer segment acquisition time than the survey scan, thereby collecting more mass spectra per segment or spectra from more ions per segment

iv) with a larger mass range than the survey scan, thereby measuring a greater number of elemental isotopic masses per segment.

18. The method of claim 17, wherein the region of interest are computationally or manually identified in the initial survey scan;

and

areas of the planar substrate that are found to be devoid of sample are omitted from subsequent imaging analyses.

19. The method of claim 15, wherein the visual image of the planar sample is reconstructed with color or shading scales based on individual or combined levels of mass-to-charge species.

20. The method of claim 15, wherein the image is displayed on a screen or an electronic file of the image with mass-to-charge information is produced.