Systems and methods for rapid scanning of images in digital microscopes

• US 10,712,548 B2
• Filed: 06/08/2017
• Issued: 07/14/2020
• Est. Priority Date: 06/08/2017
• Status: Active Grant

• Current Assignee: Microscopes International LLC
• Original Assignee: Microscopes International LLC
• Inventors: Ward, Jonathan C., Rainbolt, Richard K.
• Primary Examiner(s): Harold, Jefferey F
• Assistant Examiner(s): Sanders, Justin B
• Application Number:
  US15/616,922
• Publication Number:
  US 20180356621A1
• Time in Patent Office: 1,132 Days
• CPC Class Codes: G02B 21/086 for transillumination only
• CPC Class Codes: G02B 21/18 Arrangements with more than one light path, e.g. for comparing two specimens
• CPC Class Codes: G02B 21/241 Devices for focusing focusing in general G02B7/28
• CPC Class Codes: G02B 21/367 providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison

Abstract

Systems and methods for preparing a field image of a portion of a target area or region of interest (ROI) of a microscope slide specimen, and of assembly a plurality of field images into an image of the target area, using a SSM having a movable slide stage, an objective lens, a digital video camera and a digital image sensing element (DIS).

Claims

1. A method of preparing a field image of a portion of a target area of a microscope slide specimen using a slide scanning microscope, the slide scanning microscope having a movable slide stage for holding the microscope slide, an objective lens, and a digital video camera optically coupled to the objective lens, the digital video camera having a digital image sensor operating as an M pixels by N lines digital pixel array, the method comprising:

performing a first movement to a first stationary position, the first movement selected from a slide stage movement and an objective lens movement, the first movement having a move time and a settle time;

determining a line number S of a video frame F output by the digital video camera at the end of the settle time as a complete line;

capturing, after the settle time has elapsed, a first field image portion comprising lines S to N of video frame F output by the digital video camera for the M pixels by N lines digital pixel array;

capturing a second field image portion comprising lines 1 to (S−

1) of video frame F+1 output by the digital video camera for the M pixels by N lines digital pixel array; and

assembling the field image using the second field image portion from video frame F+1 and the first field image portion from video frame F.

2. The method of claim 1, wherein:

the moveable slide stage comprises a generally planar stage capable of holding the microscope slide;

the generally planar stage is moveable in an X-axis direction and a Y-axis direction perpendicular to the X-axis; and

performing a first movement comprises performing a slide stage movement selected from an X-axis movement performed in an X-axis move time and having an X-axis settle time, and a Y-axis movement performed in a Y-axis move time and having a Y-axis settle time.

3. The method of claim 1, wherein:

the objective lens of the slide scanning microscope is movable in a Z-axis direction generally perpendicular to the microscope slide to focus the field image at a desired focal plane within the microscope slide specimen,

performing a first movement comprises performing an objective lens movement in a Z-axis direction to focus the field image at a desired focal plane within the microscope slide specimen, the objective lens movement having a Z-axis move time and a Z-axis settle time.

4. The method of claim 3, further comprising:

a) performing another objective lens movement to focus the field image at a desired focal plane;

b) repeating said steps of;

determining a line number S of a video frame F output by the digital video camera at the end of the settle time as a complete line;

capturing, after the settle time has elapsed, a first field image portion comprising lines S to N of video frame F output by the digital video camera for the M pixels by N rows digital pixel array;

capturing a second field image portion comprising lines 1 to (S−

1) of video frame F+1 output by the digital video camera for the M pixels by N rows digital pixel array; and

assembling the field image using the second field image portion from video frame F+1 and the first field image portion from video frame F; and

c) optionally repeating steps a) and b) one or more times to obtain a plurality of field images having different focal planes.

5. The method of claim 4, further comprising:

determining a focus value for each of said plurality of field images having different focal planes; and

retaining the field image having the most appropriate focus value.

6. The method of claim 3, wherein performing an objective movement comprises moving the objective lens in a Z-axis direction while the digital image sensor and the movable stage remain stationary.

7. The method of claim 1, wherein determining a line number S occurs at a time selected from:

before the move time has elapsed;

before the settle time has elapsed; and

after the settle time has elapsed.

8. The method of claim 1 wherein determining a line number S comprises determining a line number of the first complete line of a video frame F output by the digital video camera after the end of the settle time.

9. A method of preparing an image of a target area of a microscope slide specimen from a plurality of field images, each field image comprising an image of a portion of the target area of the slide specimen, using a slide scanning microscope having a movable slide stage for holding the microscope slide, an objective lens, and a digital video camera optically coupled to the objective lens, the digital video camera having a digital image sensor operating as an M pixels by N lines digital pixel array, the method comprising:

capturing each field image of the plurality of field images by;

a) performing a first movement to a desired stationary position, the first movement selected from a slide stage movement and an objective lens movement, the first movement having a move time and a settle time;

b) determining a line number S of a video frame F output by the digital video camera at the end of the settle time as a complete line;

c) capturing, after the settle time has elapsed, a first field image portion comprising lines S to N of video frame F output by the digital video camera for the M pixels by N lines digital pixel array;

d) capturing a second field image portion comprising lines 1 to (S−

1) of video frame F+1 output by the digital video camera for the M pixels by N lines digital pixel array;

e) assembling the field image using the second field image portion from video frame F+1 and the first field image portion from video frame F;

f) repeating steps a-e for each field image in the plurality of field images; and

g) assembling the plurality of field images to form a single image of the target area.

10. The method of claim 9, wherein:

the moveable slide stage comprises a generally planar stage capable of holding the microscope slide;

the generally planar stage is moveable in an X-axis direction and a Y-axis direction perpendicular to the X-axis; and

performing a first movement comprises performing a slide stage movement selected from an X-axis movement performed in an X-axis move time and having an X-axis settle time, and a Y-axis movement performed in a Y-axis move time and having a Y-axis settle time.

11. The method of claim 9, wherein:

the objective lens of the slide scanning microscope is movable in a Z-axis direction generally perpendicular to the microscope slide to focus the field image at a desired focal plane within the microscope slide specimen, and

performing a first movement comprises performing an objective lens movement in a Z-axis direction to focus the field image at a desired focal plane within the microscope slide specimen, the objective lens movement having a Z-axis move time and a Z-axis settle time.

12. The method of claim 11, further comprising:

h) performing another objective lens movement to focus the field image at a desired focal plane;

i) repeating said steps of;

determining a line number S of a video frame F output by the digital video camera at the end of the settle time as a complete line;

capturing, after the settle time has elapsed, a first field image portion comprising lines S to N of video frame F output by the digital video camera for the M pixels by N rows digital pixel array;

capturing a second field image portion comprising lines 1 to (S−

1) of video frame F+1 output by the digital video camera for the M pixels by N rows digital pixel array; and

assembling the field image using the second field image portion from video frame F+1 and the first field image portion from video frame F; and

j) optionally repeating steps h) and i) one or more times to obtain a plurality of field images having different focal planes.

13. The method of claim 12, further comprising:

k) determining a focus value for each of said plurality of field images having different focal planes; and

l) retaining the field image having the most appropriate focus value.

14. The method of claim 11, wherein performing an objective movement comprises moving the objective lens in a Z-axis direction while the digital image sensor and the movable stage remain stationary.

16. The method of claim 12 wherein repeating at least steps a-e to capture each of the plurality of field images comprises repeating steps a-k for to capture each of the plurality of field images.

15. A method of preparing an image of a target area of a microscope slide specimen from a plurality of field images, each field image comprising an image of a portion of the target area of the slide specimen, using a slide scanning microscope having a moveable slide stage for holding the microscope slide, at least one objective lens, and a digital video camera having a digital image sensor operating as an M columns by N rows digital pixel array, the method comprising:

capturing each field image of the plurality of field images by;

a) performing a slide stage movement to move the slide stage to a desired stationary slide stage position, the slide stage movement having a move time and a settle time;

b) determining a line number S of a video frame F output by the digital video camera at the end of the settle time as a complete line;

c) capturing, after the settle time has elapsed, a first field image portion comprising lines S to N of video frame F output by the digital video camera for the M pixels by N lines digital pixel array;

d) capturing a second field image portion comprising lines 1 to (S−

1) of video frame F+1 output by the digital video camera for the M pixels by N lines digital pixel array;

e) assembling the field image using the second field image portion from video frame F+1 and the first field image portion from video frame F;

f) performing an objective lens movement to a desired stationary objective lens position, the objective lens movement having a move time and a settle time;

g) determining a line number S of a video frame G output by the digital video camera at the end of the settle time as a complete line;

h) capturing, after the settle time has elapsed, a first field image portion comprising lines S to N of video frame G output by the digital video camera for the M pixels by N lines digital pixel array;

i) capturing a second field image portion comprising lines 1 to (S−

1) of video frame G+1 output by the digital video camera for the M pixels by N lines digital pixel array;

j) repeating steps f-i to obtain a Z-stack of field images at the desired stationary slide stage position, each field image in the Z-stack of field images comprising an image having a different focus from the other images in the Z-stack;

k) retaining one field image of the Z-stack of field images as the field image for the desired stationary slide stage position;

l) repeating at least steps a-e to capture each of the plurality of field images; and

assembling the plurality of field images to form a single image of the target area.