Method and apparatus for automated assay of biological specimens

• US 5,428,690 A
• Filed: 02/18/1992
• Issued: 06/27/1995
• Est. Priority Date: 09/23/1991
• Status: Expired due to Term

• Current Assignee: Cell Analysis System Incorporated
• Original Assignee: Becton, Dickinson & Co
• Inventors: Wagner, Robert E., Bacus, James W., Bacus, James V.
• Primary Examiner(s): Mancuso, Joseph
• Assistant Examiner(s): Fox, David
• Application Number:
  US07/837,051
• Time in Patent Office: 1,225 Days
• Field of Search: 382/6, 356/39, 356/40, 364/413.07, 364/413.08, 364/413.13, 364/413.1, 422/65, 436/46, 359/391, 359/393, 358/93, 358/107
• US Class Current: 382/128
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 35/00029 provided with flat sample substrates, e.g. slides G01N35/028 takes precedence
• CPC Class Codes: G01N 35/00603 Reinspection of samples
• CPC Class Codes: G02B 21/34 Microscope slides, e.g. mounting specimens on microscope slides

Abstract

An apparatus and method for automated assay of biological specimens positioned on microscope slides. The apparatus comprises an interactive optical subsystem for viewing the biological specimen on the slide and for producing an interactive video signal corresponding to the viewed image. An automated optical subsystem includes a single high power microscope objective for scanning a rack of slides, portions of which having been previously identified for assay in the interactive optical means. The system also includes a processor for processing the interactive and automatic video signals for the two optical subsystems. The processor receives the automatic video signal and performs biological assay functions upon it. A method and apparatus are also disclosed for marking points for later analysis on the microscope slides and for associating an analysis function with each marked point.

Claims

1. A method for locating a small biological specimen on a large specimen area of a transparent slide for later automated analysis of the specimen, said method comprising the steps of:

providing a slide before an operator for viewing at low resolution at a first microscope so that the operator may rapidly exclude areas on the slide and proceed to locate an action point on a small specimen at the low resolution of a first magnification with a first microscope; and

viewing the slide at the low resolution and locating the action point in the section associated with the specimen, and storing the location as a coordinate address;

automatic scanning beginning at the coordinate address and progressing automatically through a plurality of image fields at a substantially higher magnification and resolution with a second microscope than the low resolution.

2. A method in accordance with claim 1 including the additional steps of:

providing the slide with a reference grid to divide the slide into a plurality of designated grid sections, and selecting one of the designated grid sections having specimen for viewing at the low resolution.

3. A method in accordance with claim 1 in which the step of storing the location of said specimen includes the steps of moving a mark on a monitor screen to the location of the specimen and then entering the address of the mark for initiating subsequent automatic scanning of a plurality of image fields.

4. A method in accordance with claim 1 including the additional steps of noting a previously placed mark on the slide adjacent the specimen made by a pathologist and entering the coordinate address of this previously placed mark as the address to initiate scanning at the substantially higher resolution.

5. A method in accordance with claim 1 including the step of providing a flat bed carrier and positioning the slide in the flat bed carrier with a plurality of other slides and positioning the carrier and each slide at its associated stored coordinate address to locate specimens for each of the slides to facilitate subsequent automatic scanning of a plurality of image fields of each of the slides.

36. A method in accordance with claim 2 and wherein the low resolution at which the slide is viewed provides an enlarged view of the grid section.

37. A method in accordance with claim 2 wherein the slide is viewed without magnification when selecting one of the designated grid sections having specimen.

6. A method of analysis of biological specimens on each of a plurality of slides having large specimen areas with a specimen occupying only a small fraction of the specimen area, said method comprising the steps of:

providing a plurality of specimen locating stations at which an operator locates the small specimen in the large specimen area;

providing a specimen carrier at each station for the operator to handle a plurality of slides associated with the carrier;

the operator at each station viewing a specimen slide at low resolution and the operator storing the location of the specimen on a storage media;

transferring the slides and their carrier to an automated analysis station;

transferring the stored locations of the addresses of the specimens to the analysis station;

initiating scanning of the specimens at a substantially higher resolution than the low resolution at the location address previously established by the operator; and

automatically positioning each of the slides beneath the microscope of a digital imaging apparatus for an automated analysis thereof using the previously applied addresses to avoid scanning large areas empty of the specimen to be scanned.

7. A method in accordance with claim 6 including the shifting of a flat bed carrier having a plurality of slides in front of the operator.

8. A method in accordance with claim 7 including the steps of the operator designating regions on the specimen area at a first low resolution, enlarging the designated region on the slide to show the specimen enlarged, and then noting the address to initiate scanning.

9. An interactive method of specifying one or more action points on each of a group of microscope slides mounted in a carrier for subsequent automatic image analysis comprising:

positioning a slide mounted in the carrier before an operator who visually inspects the slide to select the coordinate locations of one or more action points on the slide;

entering into storage the coordinate address of at least one action point;

selecting and entering into storage one or more appropriate actions for subsequent automatic image analysis at said at least one action point;

providing a group of slides in the carrier;

proceeding to each of the group of slides in the carrier and having the operator visually inspect each slide to select one or more action points and having the operator enter the coordinate location for each action point into storage;

forming an enlarged image on a monitor related to the location of the action point and the operator entering from the enlarged image a more finely defined coordinate address for a given action point into storage;

displaying on a monitor an image of a microscope slide having a coordinate grid pattern of rectangles thereon; and

forming a coordinate grid on at least one of the microscope slides and the operator using a display cursor to locate the action point on the monitor grid related to the location of the action point on the slide.

10. A method in accordance with claim 9 including the selecting with the cursor one of the displayed grid rectangles and then enlarging that grid rectangle for viewing on a monitor and using a spot locator to select a location spot within the enlarged grid rectangle on the monitor as the coordinate location of the action point.

11. A method of assaying with a microscope and an image analysis means of biological specimens on each of a group of microscope slides mounted in a carrier comprising:

positioning a slide before an operator who visually inspects the slide for one or more action points on the slide with a first microscope at a first resolution and selects a coordinate address on the slide for the action point;

entering into storage the address for the action point and a selected function from one of several functions performable at the various action points;

proceeding to each of the slides of the group of slides in the carrier and having the operator view each of the slides, the operator entering the coordinate address and selected analysis action for each action point into storage;

positioning the carrier with the slides therein into an operative position with respect to a second microscope at the image analysis means and at a higher resolution; and

reading each of the stored addresses for each of the slides and automatically moving the carrier relative to the second microscope to position each stored address on each slide at the microscope for performing automatically the action for that action point by the image analysis means.

12. A method in accordance with claim 11 including:

displaying an enlarged image on a monitor related to the location of the action point and the operator entering from the enlarged image a more finely defined coordinate address for a given action point into storage.

13. A method in accordance with claim 12 including the steps of locating an action point for light calibration and locating an action point for analysis of specimen cells.

14. A method in accordance with claim 13 including the forming of a grid on the slide and for the operator viewing a grid displayed on the monitor from storage, the operator using a cursor to locate the address point on the grid on the monitor related to the location of the action point on the slide.

15. A method in accordance with claim 14 including the selecting with the cursor one of the grid areas and then enlarging that grid area for viewing on a monitor and using a spot locator to select a location spot within the enlarged grid area on the monitor as the coordinate address of the action point.

16. An apparatus for interactively specifying one or more action points on each of a group of microscope slides, said apparatus comprising:

means for holding the group of slides in a side-by-side relationship before an operator who visually inspects the slides for one or more action points on each slide;

means for providing an associated visible coordinate system for the slide to allow an operator to select a coordinate address on the slide for each action point;

a monitor to display the visible coordinate system with respect to a slide at the operative position;

a storage means for storing addresses of selected action points on the slide;

means for displaying an enlarged image on the monitor related to the location of the action point at a resolution substantially lower than the resolution at which said action point is subsequently analyzed; and

means for generating a spot on the enlarged image and thereby to produce a more finely defined coordinate address for a given action point;

wherein said visible coordinate system is provided by means for generating a grid pattern on the slide for viewing by the operator, and wherein said apparatus includes a monitor having a grid displayed from storage, a monitor cursor to locate the address point on the grid on the monitor related to the location of the action point on the slide.

17. An apparatus in accordance with claim 16 including means for producing from storage an enlarged view of a grid area on the monitor and a spot generator to selected a location spot within the enlarged grid area on the monitor as the coordinate address.

18. An apparatus for assaying biological specimens on each of a group of microscope slides, said apparatus comprising:

means for holding a slide before an operator who visually inspects the slide for one or more action points on the slide;

means providing an associated coordinate system for the slide to allow an operator to select a coordinate address on the slide for the action point;

a storage means for storing in the storage means address and associated analysis action for each slide;

means for entering into storage the address and a selected analysis action from one of several analysis actions performable at the various action points;

said means for holding slide, holding the next slide of the group of slides in the carrier for viewing the next slide and an associated coordinate system provided therewith and selecting an action point thereon, said means for entering being operable by the operator to enter the coordinate address and selected analysis action for each action point into storage;

an image analysis means for receiving a carrier with the slides therein into an operative position;

a microscope in the image analysis means for viewing a slide at the operative position;

means for automatically moving the carrier relative to the microscope to position each stored address on each slide at the microscope operative position for performing the associated stored analysis action for that action point by the image analysis means;

a monitor to display the coordinate system with respect to a slide at the operative position;

means for forming an enlarged image on the monitor related to the location of the action point;

means for generating a spot on the enlarged image and thereby to produce a more finely defined coordinate address for a given point; and

means for generating a grid pattern on the slide for viewing by the operator, said monitor having a grid displayed from storage, and a cursor to locate the address point on the grid on the monitor related to the location of the action point on the slide.

19. An apparatus in accordance with claim 18 in which the image analysis means includes means for performing a light calibration action at one addressed action point on a slide and means for analyzing specimen cells at another addressed action point on a slide.

20. An apparatus in accordance with claim 18 including means for producing from storage an enlarged view of a grid area on the monitor and a spot generator to select a location spot within the enlarged grid area on the monitor as the coordinate address of an action point.

21. An apparatus for automated assay of biological specimens positioned on microscope slides comprising:

a first microscope of a first resolution for viewing action points on a microscope slide;

specifying means actuated by an operator for specifying at least one action point on a microscope slide by an operator;

selecting means operated by an operator for selecting an analysis action from one of several actions performable by said automated apparatus to be performed at each action point specified by the specifying means;

means operable by said specifying means and selecting means for storing associated representations of the location of each specified action point and each defined action to be performed at the specified action point for subsequent automatic analysis;

means for reading a specified action point representation and the defined action representation associated therewith from the storage means; and

automatic analysis means including a second microscope having a microscope objective at a higher resolution, for aligning said microscope objective with each action point represented by the action point representation read from said storage means and for performing the action represented by the action representation associated therewith.

22. An apparatus in accordance with claim 21 wherein said means for specifying comprises display means for displaying a visual image of portions of a microscope slide;

means responsive to operator interaction for moving a cursor on said displayed visual image; and

means for selecting an action location identified by said cursor.

23. An apparatus in accordance with claim 22 wherein said display means displays location reference markings on the displayed visual image.

24. An automated apparatus in accordance with claim 23 comprising means for forming on the microscope slide a visual pattern of location reference markings substantially identical to the pattern of reference markings displayed on said displayed visual image.

25. An apparatus in accordance with claim 22 wherein said display means displays a pattern of grid markings defining a plurality of rectangles on said displayed visual image, and said means for specifying comprises means for specifying one of said rectangles.

26. An automated apparatus in accordance with claim 25 comprising means for forming on the microscope slide a pattern of grid markings substantially identical to the grid markings presented on said displayed visual image.

27. An apparatus in accordance with claim 21 wherein said means for defining comprises display means for displaying a plurality of possible actions;

means responsive to operator interaction for moving a cursor on the displayed plurality of actions; and

means for selecting one of said plurality of actions identified by said cursor.

28. An automated apparatus in accordance with claim 22 comprising means for displaying indicia of an identified action point on said displayed visual image of portions of a microscope slide.

29. An apparatus in accordance with claim 21 wherein one of said definable actions comprises a scan action and said means for defining actions comprises means for accumulating information identifying a type of analysis to be performed from a plurality of types of analysis performable by said automatic analysis means, and said means for storing, stores the information identifying the type of analysis in association with a specified action point.

30. An apparatus in accordance with claim 29 wherein said type of analysis includes an estrogen/progesterone assay.

31. An apparatus in accordance with claim 29 wherein said type of analysis includes a DNA mass assay.

32. An apparatus in accordance with claim 21 wherein one of said definable actions comprises a scan action, said means for defining an action comprises means for accumulating information describing a pattern of scan movements between said microscope slide and said microscope objective, and said means for storing stores the information describing the pattern of scan movements in association with a specified action point.

33. An automated apparatus for identifying action points for the performance of analysis functions on biological specimens positioned on microscope slides comprising;

means operable manually by an operator for specifying by operator interaction at least one action point address on a microscope slide;

means operable manually by an operator for selecting an analysis action from one of several actions performable by said automated apparatus to be performed at each action point address specified by the operator operating said means for specifying;

means for storing in a data file associated representations of each specified action point address and each defined action to be performed at the specified action point;

means for transmitting to a separate image analysis apparatus said data file for performance of said specified actions;

means responsive to operator interaction for moving a cursor on said displayed visual image;

means for selecting an action point identified by said cursor;

wherein said display means displays location reference markings on the displayed visual image; and

means for forming on the microscope slide a visual pattern of location reference markings substantially identical to the pattern of reference markings on said displayed visual image.

38. An apparatus in accordance with claim 33 wherein said means for defining comprises display means for displaying a plurality of possible actions;

means responsive to operator interaction for moving a cursor on the displayed plurality of actions; and

means for selecting one of said plurality of actions identified by said cursor.

34. An automated apparatus for identifying action points for the performance of analysis functions on biological specimens positioned on microscope slides comprising:

means operable manually by an operator for specifying by operator interaction at least one action point address on a microscope slide;

means operable manually by an operator for selecting an analysis action from one of several actions performable by said automated apparatus to be performed at each action point address specified by the operator operating said means for selecting;

means for storing in a data file associated representations of each specified action point address and each defined action to be performed at the specified action point;

means for transmitting separate image analysis apparatus said data file for performance of said specified actions;

means responsive to operator interaction for moving a cursor on said displayed visual image;

means for selecting an action point identified by said cursor;

wherein said display means displays a pattern of grid markings defining a plurality of rectangles on said displayed visual image, and said means for specifying comprises means for specifying one of said rectangles; and

means for forming on the microscope slide a pattern of grid markings substantially identical to the grid markings presented on said displayed visual image.

35. An apparatus in accordance with claim 34 wherein said means for defining comprises display means for displaying a plurality of possible actions;

means responsive to operator interaction for moving a cursor on the displayed plurality of actions; and

means for selecting one of said plurality of actions identified by said cursor.