Method and apparatus for automated image analysis of biological specimens

• US 20070053569A1
• Filed: 11/07/2006
• Published: 03/08/2007
• Est. Priority Date: 11/30/1995
• Status: Active Grant

• Current Assignee: Carl Zeiss Microscopy GmbH (Carl-Zeiss-Stiftung)
• Original Assignee: CARL Zeiss Microimaging GmbH (Carl-Zeiss-Stiftung)
• Inventors: Riding, Thomas, Douglass, James, Ring, James
• Granted Patent: US 7,359,548 B2
• Time in Patent Office: Days
• US Class Current: 382/128
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1468 with spatial resolution of the texture or inner structure of the particle
• CPC Class Codes: G01N 2015/1472 with colour
• CPC Class Codes: G01N 2015/1488 Methods for deciding
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G02B 21/244 using image analysis techniques
• 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
• CPC Class Codes: G06T 7/60 Analysis of geometric attributes
• CPC Class Codes: G06V 20/69 Microscopic objects, e.g. biological cells or cellular parts
• CPC Class Codes: G06V 20/693 Acquisition

Abstract

A method and apparatus for automated cell analysis of biological specimens automatically scans at a low magnification to acquire images which are analyzed to determine candidate cell objects of interest. The low magnification images are converted from a first color space to a second color space. The color space converted image is then low pass filtered and compared to a threshold to remove artifacts and background objects from the candidate object of interest pixels of the color converted image. The candidate object of interest pixels are morphologically processed to group candidate object of interest pixels together into groups which are compared to blob parameters to identify candidate objects of interest which correspond to cells or other structures relevant to medical diagnosis of the biological specimen. The location coordinates of the objects of interest are stored and additional images of the candidate cell objects are acquired at high magnification. The high magnification images are analyzed in the same manner as the low magnification images to confirm the candidate objects of interest which are objects of interest. A high magnification image of each confirmed object of interest is stored for later review and evaluation by a pathologist.

Claims

1. An electronic microscope system, comprising:

a processor;

a movable stage, movable in two orthogonal directions, under control of the processor, said movable stage includes at least one part that determines a current position of the movable stage;

a microscope part, which magnifies a sample on the movable stage;

an electronic camera part, which obtains an image of a magnified sample at a specified position controlled by said movable stage; and

at least one vibration isolation mount, coupled to said stage, and isolating vibration between said movable stage and said microscope part.

2. A system as in claim 1, wherein said vibration isolation mount includes a high viscosity silicon gel.

3. A system as in claim 1, wherein said vibration isolation mount includes a spring and piston.

4. A system as in claim 1, wherein said vibration isolation mount obtains damping factors on the order of 17 to 20%.

5. A system as in claim 1, wherein said movable stage is movable in x and y directions.

6. A system as in claim 5, further comprising an x-position motor, moving the movable stage in an x direction, and a y position motor, moving the stage in the y direction.

7. A system as in claim 5, wherein the processor controls storage of electronic images along with information about x,y positions of the stage at the time when the electronic images were obtained.

8. A method comprising:

using a processor to move a sample holding stage;

magnifying an image of an area of the sample on the movable stage, where the area is based on a moved position of the sample holding stage;

obtaining an electronic file indicative of the magnified image; and

isolating vibration between said moving of said stage and said magnifying and obtaining.

9. A method as in claim 8, wherein said isolating comprises using a high viscosity silicon gel to isolate said vibration.

10. A method as in claim 8, wherein said isolating comprises using a spring and piston for isolating the vibration.

11. A method as in claim 8, wherein said isolating comprises obtaining damping factors on the order of 17 to 20%.

12. A method as in claim 8, wherein said using comprises moving the stage in X and y directions.

13. A method as in claim 12, wherein said moving comprises controlling separate motors where an x-position motor is controlled to move the movable stage in an x direction, and a y position motor is controlled to move the stage in the y direction.

14. A method as in claim 12, further comprising storing information about x,y positions of the stage at the time when the electronic files were obtained.

15. A system, comprising:

an electronic microscope that obtains an electronic signal indicative of pixels of a microscope image; and

a processor, that processes said electronic signal to determine relationships between different colors within the whole electronic signal.

16. A system as in claim 15, wherein said processor calculates ratios between different color pixels of the signal.

17. A system as in claim 15, wherein said processor calculates ratios between a first channel of color of the image across the entire image, and a second channel of color of the image across the entire image.

18. A system as in claim 17, further comprising calculating a color balance for the image.

19. A system as in claim 15, wherein said processor determines a relationship between a first color channel and a second color channel.

20. A system as in claim 19, wherein said first and second color channels are each one of red, green and blue channels.

21. A system as in claim 19, wherein said relationship is a ratio.

22. A system as in claim 19, wherein said processor calculates at least one ratio from the group consisting of red/red;

red/green;

red/blue;

green/green;

green/blue;

green/red;

blue/blue;

blue/green;

blue/red.

23. A system as in claim 19 wherein said processor calculates ratios of each of red/red;

red/green;

red/blue;

green/green;

green/blue;

green/red;

blue/blue;

blue/green;

blue/red.

24. A method, comprising:

obtaining an electronic signal indicative of pixels of a microscope image; and

processes said electronic signal to determine relationships between different colors within each of the pixels of the image.

25. A method as in claim 24, further comprising calculating ratios between different color pixels of the image.

26. A method as in claim 24, further comprising calculating ratios between a first channel of color of the image across the entire image, and a second channel of color of the image across the entire image.

27. A method as in claim 25, further comprising calculating a color balance for the image.

28. A method as in claim 24, further comprising calculating a relationship between a first color channel and a second color channel.

29. A method as in claim 28, wherein said first and second color channels are each one of red, green and blue channels.

30. A method as in claim 24, wherein said relationship is a ratio.

31. A method, comprising:

obtaining an electronic version of a microscope image; and

determining information about colors within the image by establishing a high threshold and a low threshold for a specified color, counting a number of pixels that are in between said high threshold and said low threshold, dividing the number of pixels by a value which relates pixels to cell size, and using a resultant number as a count of the number of cells within the image.

32. A method as in claim 31, wherein said colors are colors of a stain on a slide.

33. A method as in claim 32, wherein said stain is hematoxylin, and said colors are a blue color matching the characteristics of hematoxylin.

34. A method, comprising:

staining a biological sample with a stain of a specified color;

obtaining an electronic version of a microscope image of said biological sample;

using said specified color to determine high threshold and a low threshold around said specified color;

analyzing the image, to determine information about areas in the image that are in between said high threshold and said low threshold; and

using said information to determine a count of the number of stained cells in the image.

35. A method as in claim 34, wherein said using comprises, determining a number of pixels which are between said thresholds, dividing the number of pixels by a value which relates pixels to cell size, and using a resultant number as a count of the number of cells within the image.