Method and apparatus for acquiring and reconstructing magnified specimen images from a computer-controlled microscope

• US 6,226,392 B1
• Filed: 09/13/1999
• Issued: 05/01/2001
• Est. Priority Date: 08/23/1996
• Status: Expired due to Term

• Current Assignee: Evident Scientific Incorporated (Olympus Corporation)
• Original Assignee: Bacus Laboratories Incorporated (Olympus Corporation)
• Inventors: Bacus, James W., Bacus, James V.
• Primary Examiner(s): Johns, Andrew W.
• Assistant Examiner(s): Le, Hieu C.
• Application Number:
  US09/395,694
• Time in Patent Office: 596 Days
• Field of Search: 382/240, 382/224, 382/128, 382/284, 382/294, 382/133, 348/79, 356/372, 356/39, 600/407, 250/455, 345/435, 128/922, 364/413.07, 364/413.13
• US Class Current: 382/128
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G06T 3/4038 Image mosaicing, e.g. composing plane images from plane sub-images
• CPC Class Codes: G06T 7/0012 Biomedical image inspection
• CPC Class Codes: G06V 20/695 Preprocessing, e.g. image segmentation
• CPC Class Codes: Y10S 128/922 including image analysis

Abstract

A method is disclosed for using a computer controlled microscope system to provide a reconstructed, seamless image from several contiguous fields of view to show the architecture and spatial relationship of biological material in a specimen. The specimen is scanned with a microscope and a digital scanner to provide digitized titles of contiguous, fields of view at a predetermined magnification, optical resolution and pixel resolution. Preferably, an automated X, Y stage with higher positional spatial resolution than the spatial pixel resolution of the digital scanner is used to acquire the contiguous fields of view and the image registration information which is used to reconstruct and to display on a monitor the reconstructed image formed of the contiguous multiple fields of view. Preferably, a macro image of contiguous image tiles at a low magnification and optical resolution is obtained and the user navigates within the macro image and selects areas. Higher magnification titles for the selected area are assembled and viewed at a higher magnification and optical resolution as a composite micro image of the selected area. Both the macro and micro images may be displayed simultaneously. Preferably, the user may scroll and cause additional fields of view to be added to previously displayed fields of view.

Claims

1. A method of using a computer controlled microscope imaging system to provide a reconstructed seamless image reconstructed from several contiguous fields of view showing the architecture and spatial relationship of biological material in a biological specimen, the method comprising:

scanning the specimen with a microscope at a predetermined magnification, optical resolution and pixel resolution and acquiring digitized image tiles of contiguous fields of view;

acquiring image registration information for the image tiles using microscope stage coordinates;

assembling and coordinating the digitized image tiles using a acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed seamless composite image formed of a plurality of digitized, contiguous image tiles;

displaying the reconstructed, seamless, composite image showing the architecture and the spatial relationship of the biological material in contiguous image tiles at an optical resolution for viewing and diagnostic analysis without the aid of a microscope;

scanning the specimen at a first prior time at a lower magnification and optical resolution and acquiring contiguous image tiles and acquiring image registration information for each of these lower magnification, image tiles to align the respective edges thereof;

assembling and coordinating the digitized, lower magnification, image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed, seamless, composite, lower magnification image formed of a plurality of digitized, contiguous, image tiles; and

navigating within the specimen'"'"'s composite, lower magnification and optical resolution image and selecting adjacent segments of the navigated area on the composite, overall specimen image for the tile image acquisition at a predetermined, higher predetermined magnification and optical resolution.

2. A method of using a computer controlled microscope imaging system to provide a reconstructed seamless image reconstructed from several contiguous fields of view showing the architecture and spatial relationship of biological material in a biological specimen, the method comprising:

scanning the specimen with a microscope at a predetermined magnification, optical resolution and pixel resolution and acquiring digitized image tiles of contiguous fields of view;

acquiring image registration information for the image tiles using microscope stage coordinates;

assembling and coordinating the digitized image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed seamless composite image formed of a plurality of digitized, contiguous image tiles;

displaying the reconstructed, seamless, composite image showing the architecture and the spatial relationship of the biological material in contiguous image tiles at an optical resolution for viewing and diagnostic analysis without the aid of a microscope; and

scrolling by the user and displaying additional adjacent image tiles to the previously displayed fields of view to create a new composite image and to bring into view a further architecture and spatial relationship including the biological material displayed before and after scrolling the optical resolution for diagnostic analysis.

3. A method of using a computer controlled microscope imaging system to provide a reconstructed seamless image reconstructed from several contiguous fields of view showing the architecture and spatial relationship of biological material in a biological specimen, the method comprising:

scanning the specimen with a microscope at a predetermined magnification, optical resolution and pixel resolution and acquiring digitized image tiles of contiguous fields of view;

acquiring image registration information for the image tiles using microscope stage coordinates;

assembling and coordinating the digitized image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed seamless composite image formed of a plurality of digitized, contiguous image tiles;

scanning the specimen at a first prior time at a lower magnification and optical resolution and acquiring contiguous image tiles and acquiring image registration information for each of these lower magnification, image tiles to align the respective edges thereof;

assembling and coordinating the digitized, lower magnification, image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed, seamless, composite, lower magnification image formed of a plurality of digitized, contiguous, image tiles; and

displaying the reconstructed, seamless, composite image showing the architecture and the spatial relationship of the biological material in contiguous image tiles at an optical resolution for viewing and diagnostic analysis without the aid of a microscope;

the scanning of the specimen at the predetermined magnification and optical resolution being done subsequent to the scanning of the specimen with a microscope at a lower magnification and optical resolution; and

displaying each of the seamless, composite images at different magnifications and optical resolutions simultaneously to the viewer.

4. A method in accordance with claim 3 wherein:

the scanning of the specimen with a microscope at first and second magnifications and optical resolutions is done at a first location;

transmitting the digitized and stored contiguous fields of view and edge registering information to a remote location;

displaying each of the reconstructed, seamless, composite images simultaneously to the user; and

interaction by the user with the composite, lower magnification seamless image at the remote location to provide new displays of seamless, reconstructed, composite images at the higher resolution to the remote viewer of adjacent segments of specimen.

5. A method of using a computer controlled microscope imaging system to provide a reconstructed seamless image reconstructed from several contiguous fields of view showing the architecture and spatial relationship of biological material in a biological specimen, the method comprising:

scanning the specimen with a microscope at a predetermined magnification, optical resolution and pixel resolution and acquiring digitized image tiles of contiguous fields of view;

acquiring image registration information for the image tiles using microscope stage coordinates;

assembling and coordinating the digitized image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed seamless composite image formed of a plurality of digitized, contiguous image tiles;

scanning the specimen at a first prior time at a lower magnification and optical resolution and acquiring contiguous image tiles and acquiring image registration information for each of these lower magnification, image tiles to align the respective edges thereof;

assembling and coordinating the digitized, lower magnification, image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed, seamless, composite, lower magnification image formed of a plurality of digitized, contiguous, image tiles;

displaying the reconstructed, seamless, composite image showing the architecture and the spatial relationship of the biological material in contiguous image tiles at an optical resolution for viewing and diagnostic analysis without the aid of a microscope; and

forming a composite, magnified and reconstructed image at the lower magnification and lower resolution for displaying substantially the entire specimen to the observer.

6. A method of using a computer controlled microscope imaging system to provide a reconstructed seamless image reconstructed from several contiguous fields of view showing the architecture and spatial relationship of biological material in a biological specimen, the method comprising:

scanning the specimen with a microscope at a predetermined magnification, optical resolution and pixel resolution and acquiring digitized image tiles of contiguous fields of view;

acquiring image registration information for the image tiles using microscope stage coordinates;

assembling and coordinating the digitized image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed seamless composite image formed of a plurality of digitized, contiguous image tiles;

displaying the reconstructed, seamless, composite image showing the architecture and the spatial relationship of the biological material in contiguous image tiles at an optical resolution for viewing and diagnostic analysis without the aid of a microscope;

using an automated X, Y stage with higher positional spatial resolution than the spatial pixel resolution of a digital image scanner used to acquire the contiguous fields of view; and

using the higher spatial resolution X, Y stage coordinates associated with each image tile as the registration information to reconstruct and display on a display monitor a visually seamless, reconstructed image formed of the contiguous, multiple fields of view.

7. The method of claim 6 including using stage-positional, spatial resolution of at least about 0.1 micron or less when acquiring image registration information to align respective edges of contiguous image tiles.

8. The method of claim 7 comprising using a closed loop stage drive system to provide the at least about 0.1 micron stage-positional, spatial resolution.

9. A computer controlled microscope imaging system to provide a reconstructed seamless image reconstructed from several contiguous fields of view showing the architecture and spatial relationship of biological material in a biological specimen, the system comprising:

a microscope for scanning the specimen at a predetermined magnification, optical resolution and pixel resolution for acquiring contiguous fields of view of the specimen;

an imaging subsystem including a digital image scanner connected to the microscope for acquiring digitized image tiles of contiguous fields of view, and for acquiring image registration information for the image tiles using microscope stage coordinates, and for assembling and coordinating the digitized image tiles using the acquired image registration information to align respective edges of the contiguous image tiles to form a reconstructed seamless composite image formed of a plurality of digitized, contiguous image tiles; and

a display screen device for displaying the reconstructed, seamless, composite image showing the architecture and the spatial relationship of the biological material in contiguous image tiles at an optical resolution for viewing and diagnostic analysis without the aid of a microscope;

an automated X, Y stage with higher positional, spatial resolution than the spatial pixel resolution of the digital image scanner used to acquire the contiguous fields of view; and

the display screen device using the higher spatial resolution X, Y stage coordinates associated with each image tile as the registration information to reconstruct and to display a visually seamless, reconstructed image formed of the contiguous, multiple filed of views.

10. The system of claim 9 wherein the automated X, Y stage has a positional, spatial resolution of at least about 0.1 micron or less when acquiring image registration information to align respective edges of contiguous image tiles.

11. The system of claim 10 wherein the X, Y stage comprises a closed loop stage drive system to provide the at least about 0.1 micron stage-positional, spatial resolution.