# US 12,665,165 B2 · Method for preparing a microscopic sample for FIB/SEM tomography

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- Number: US 12,665,165 B2
- Document ID: US12665165B2
- Status: Granted
- Kind: B2
- Application number: 18/480,012
- Filed: 2023-10-03
- Published: 2024-04-11
- Granted: 2026-06-23
- Earliest claimed: 2022-10-06
- Assignees: Carl Zeiss Microscopy GmbH
- Applicants: Carl Zeiss Microscopy GmbH
- Inventors: Lorenz Lechner
- Web page: https://patentgrep.com/patent/US12665165B2

## CPC

- G01N 1/286: {involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising (microtomes G01N1/06; pulverising in general B02C; mixing in general B01F)}
- H01J 37/3056: {for microworking, e. g. etching of gratings or trimming of electrical components}
- G01N 2001/2873: {Cutting or cleaving}
- G01N 2001/2886: {Laser cutting, e.g. tissue catapult}
- H01J 2237/221: Image processing
- H01J 2237/31745: for preparing specimen to be viewed in microscopes or analyzed in microanalysers
- H01J 2237/31749: Focused ion beam
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- H01J: ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS (spark-gaps H01T; electric arc lamps H05B31/00; particle accelerators H05H)

## Abstract

A method for preparing a volume of interest from a bulk sample uses a particle beam system configured to accommodate and to image the bulk sample and configured to provide a particle beam for the removal of material from the bulk sample. Moreover, the particle beam system is configured to define trimming regions. A trimming region determines location and size of a volume to be removed from the bulk sample.

## Claims

### Claim 1

A method of exposing a volume of interest (VOI) present in a bulk sample, the method comprising:
arranging the bulk sample in a first orientation relative to an optical axis of a particle beam system configured to provide a particle beam along its optical axis;
recording an image of the bulk sample while the bulk sample is in the first orientation;
based on the first image, defining a first trimming region which determines a location and a size of a first volume of the bulk sample;
using the particle beam to remove the first volume from the bulk sample to expose a first surface of the VOI;
arranging the bulk sample in a second orientation relative to the optical axis of the particle beam system, the second orientation being different from the first orientation;
recording a second image of the bulk sample while the bulk sample is in the second orientation;
based on the second image of the bulk sample, defining a second trimming region which determines a location and a size of a second volume of the bulk sample;
using the particle beam to remove the second volume from the bulk sample to expose a second surface of the VOI, the second surface being different from the first surface, the second surface directly contacting the first surface;
based on the second image, defining a third trimming region which defines a location and a size of a third volume of the bulk sample; and
using the particle beam to remove the third volume from the bulk sample to expose a third surface of the VOI, the third surface being different from both the first and second surfaces, the third surface directly contacting the first surface.

### Claim 2 (depends on 1)

The method of claim 1, further comprising, after removing the first, second and third volumes of the bulk sample, tomographically analyzing the VOI.

### Claim 3 (depends on 2)

The method of claim 2, wherein removing the first volume from the bulk sample exposes a cross-section plane that is parallel to the optical axis when the bulk sample is arranged in its first orientation.

### Claim 4 (depends on 3)

The method of claim 3, wherein, when the bulk sample is in its second orientation, the cross-section plane faces the particle beam.

### Claim 5 (depends on 4)

The method of claim 4, wherein the second volume is adjacent a first side of the VOI, the third volume is adjacent a second side of the VOI, and the first and second sides of the VOI are opposite sides of the VOI.

### Claim 6 (depends on 5)

The method of claim 5, wherein the particle beam system comprises a rotation axis that allows arranging the bulk sample in the first orientation and in the second orientation by rotating the bulk sample around the rotation axis.

### Claim 7 (depends on 1)

The method of claim 1, wherein removing the first volume from the bulk sample exposes a cross-section plane that is parallel to the optical axis when the bulk sample is arranged in its first orientation.

### Claim 8 (depends on 1)

The method of claim 1, wherein:
removing the first volume from the bulk sample exposes a cross-section of the bulk sample; and
when the bulk sample is in its second orientation, the cross-section of the bulk sample faces the particle beam.

### Claim 9 (depends on 1)

The method of claim 1, wherein the second volume is adjacent a first side of the VOI, the third volume is adjacent a second side of the VOI, and the first and second sides of the VOI are opposite sides of the VOI.

### Claim 10 (depends on 1)

The method of claim 1, wherein the particle beam system comprises a rotation axis that allows arranging the bulk sample in the first orientation and in the second orientation by rotating the bulk sample around the rotation axis.

### Claim 11 (depends on 10)

The method of claim 10, wherein the bulk sample is rotated by 90° around the rotation axis to move the bulk sample from the first orientation to the second orientation, and the bulk sample is rotated by −90° around the rotation axis to move from the second orientation to the first orientation.

### Claim 12 (depends on 1)

The method of claim 1, wherein the particle beam system comprises a FIB system.

### Claim 13 (depends on 1)

The method of claim 1, wherein the particle beam system comprises a laser system.

### Claim 14

One or more machine-readable hardware storage devices comprising instructions that are executable by one or more processing devices of a particle beam system to perform operations comprising:
arranging a bulk sample in a first orientation relative to an optical axis of the particle beam system configured to provide a particle beam along its optical axis, a volume of interest (VOI) being present in the bulk sample;
recording an image of the bulk sample while the bulk sample is in the first orientation;
based on the first image, defining a first trimming region which determines a location and a size of a first volume of the bulk sample;
using the particle beam to remove the first volume from the bulk sample to expose a first surface of the VOI;
arranging the bulk sample in a second orientation relative to the optical axis of the particle beam system, the second orientation being different from the first orientation;
recording a second image of the bulk sample while the bulk sample is in the second orientation;
based on the second image of the bulk sample, defining a second trimming region which determines a location and a size of a second volume of the bulk sample;
using the particle beam to remove the second volume from the bulk sample to expose a second surface of the VOI, the second surface being different from the first surface, the second surface directly contacting the first surface;
based on the second image, defining a third trimming region which defines a location and a size of a third volume of the bulk sample; and
using the particle beam to remove the third volume from the bulk sample to expose a third surface of the VOI, the third surface being different from both the first and second surfaces, the third surface directly contacting the first surface.

### Claim 15

A particle beam system comprising:
one or more processing devices; and
one or more machine-readable hardware storage devices comprising instructions that are executable by the one or more processing devices to perform operations comprising:
arranging a bulk sample in a first orientation relative to an optical axis of the particle beam system configured to provide a particle beam along its optical axis, a volume of interest (VOI) being present in the bulk sample;
recording an image of the bulk sample while the bulk sample is in the first orientation;
based on the first image, defining a first trimming region which determines a location and a size of a first volume of the bulk sample;
using the particle beam to remove the first volume from the bulk sample to expose a first surface of the VOI;
arranging the bulk sample in a second orientation relative to the optical axis of the particle beam system, the second orientation being different from the first orientation;
recording a second image of the bulk sample while the bulk sample is in the second orientation;
based on the second image of the bulk sample, defining a second trimming region which determines a location and a size of a second volume of the bulk sample;
using the particle beam to remove the second volume from the bulk sample to expose a second surface of the VOI, the second surface being different from the first surface, the second surface directly contacting the first surface;
based on the second image, defining a third trimming region which defines a location and a size of a third volume of the bulk sample; and
using the particle beam to remove the third volume from the bulk sample to expose a third surface of the VOI, the third surface being different from both the first and second surfaces, the third surface directly contacting the first surface.