U.S. patent application number 09/752022 was filed with the patent office on 2001-12-13 for method and apparatus for acquiring and reconstructing magnified specimen images from a computer-controlled microscope.
This patent application is currently assigned to Bacus Research Laboratories, Inc.. Invention is credited to Bacus, James V., Bacus, James W..
Application Number | 20010050999 09/752022 |
Document ID | / |
Family ID | 27015227 |
Filed Date | 2001-12-13 |
United States Patent
Application |
20010050999 |
Kind Code |
A1 |
Bacus, James V. ; et
al. |
December 13, 2001 |
Method and apparatus for acquiring and reconstructing magnified
specimen images from a computer-controlled microscope
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.
Inventors: |
Bacus, James V.; (Downers
Grove, IL) ; Bacus, James W.; (Oakbrook, IL) |
Correspondence
Address: |
FITCH EVEN TABIN AND FLANNERY
120 SOUTH LA SALLE STREET
SUITE 1600
CHICAGO
IL
606033406
|
Assignee: |
Bacus Research Laboratories,
Inc.
|
Family ID: |
27015227 |
Appl. No.: |
09/752022 |
Filed: |
December 28, 2000 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09752022 |
Dec 28, 2000 |
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09395694 |
Sep 13, 1999 |
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6226392 |
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Current U.S.
Class: |
382/128 ;
382/299 |
Current CPC
Class: |
G06T 7/0012 20130101;
G16H 30/40 20180101; G06T 3/4038 20130101; G16H 10/40 20180101;
G16H 30/20 20180101; G01N 15/1475 20130101; G16H 40/63 20180101;
G06V 20/69 20220101 |
Class at
Publication: |
382/128 ;
382/299 |
International
Class: |
G06K 009/00; G06K
009/32 |
Claims
What is claimed is:
1. A method of using a computer-controlled microscope imaging
system for acquiring and analyzing areas of interest, said method
comprising the steps of: scanning and digitizing a specimen at a
low magnification through a microscope; displaying to the observer
a low magnification, digitized image of the specimen; selecting a
segment of the specimen from the low magnification digitized image
for viewing at a higher magnification greater than the low
magnification; automatically scanning the selected segment at the
higher magnification and digitizing the image of the segment; and
making available in real time to the view of the observer the
respective, low magnification, digitized image and the high
magnification, digitized image of the segment.
2. The method of claim 1 wherein the step of scanning and
digitizing at low magnification further includes the steps of:
positioning and coordinating spatially a plurality of low
magnification image tiles spatially to provide a composite macro
image of the specimen formed from the plurality of low
magnification image tiles; and providing a macro image of the
specimens having a field of view substantially larger than the
field of view yielded by the objective lens used for the low
magnification scanning.
3. A method in accordance with claim 2 including the step of
reducing the size of the composite, macro image before displaying
the composite macro image of the specimen to the viewer.
4. A method in accordance with claim 2 wherein the step of
selecting and digitizing the low magnification, digitized image
further includes the step of positioning and coordinating spatially
a plurality of low magnification images spatially to provide a
composite image having a field of view substantially larger than
the field of view yielded by the objective lens used for high
magnification scanning.
5. A method in accordance with claim 1 in which the step of the
selecting a defined segment area of interest further includes the
step of moving a marker on the low magnification image to a point
of interest to be viewed at high magnification; and operating the
microscope to change objectives and conditions of illumination to
provide a good image at high magnification.
6. A method in accordance with claim 5 wherein the high and low
magnification images are each reconstructed from image tiles each
having a spatially known location used to assure their proper
positioning in the reconstructed image.
7. A method in accordance with claim 6 wherein the step of
automatically changing from one magnification scanning operation to
another magnification scanning operation which includes one or more
of the steps of: changing the microscope objective lens to a
different magnification; changing at least one filter; changing the
field diaphragm; changing the light intensity; changing the focus;
and shifting the microscopic stage.
8. The method of claim 5 including the step of displaying the low
magnification image of the specimen and displaying the marker on
the low magnification image, and simultaneously displaying the high
magnification image of the segment at a position located by the
marker on the low magnification image.
9. The method of claim 1 including the step of shifting the marker
to different segments and viewing at high magnification each of the
selected segments; and marking on the low magnification image, each
of the segments viewed at high magnification.
10. The method of claim 1 including the steps of locating the
microscope on the Internet having a web site therefor; including
the step of connecting an observer's computer and viewing screen
over the Internet to the computer-controlled microscope at the web
site in real time and transmitting the digitized signals for low
magnification image and high magnification image from the web site
to the observer's computer and viewing screen.
11. The method of claim 1 including the step of enhancing the
reconstructed, high magnification image by color filtering.
12. The method of claim 1 including the step of providing a full
color, low magnification, digitized image to the observer for
aiding the observer in selecting a plurality of areas of interest
on a priority basis.
13. The method of claim 1 including the steps of: analyzing the
specimen on the slide with image analysis tests; and reporting the
results of the tests to the observer while at least one of the
respective images is being viewed by the observer.
14. The method of claim 13 wherein the specimen is a biological
specimen, and the analyzing step including performing analysis
tests on tissue or cells in the biological specimen.
15. A method in accordance with claim 1 including the step of the
observer changing the magnification of the high magnification image
to a magnification intermediate the high and low magnification
images by the use of an image analysis algorithm.
16. A method in accordance with claim 1 including the step of
storing high magnification image tiles of adjacent sections of the
specimen; and scrolling the high magnification image tiles to bring
into view on a screen adjacent sections not previously viewed.
17. A method of acquiring and analyzing biological specimens with a
computer-controlled microscope, said method comprising the steps
of: scanning and digitizing a series of fields of views through a
microscope objective lens of a low magnification and providing a
series of digitized fields of view images; arranging and
positioning the adjacent fields of view images and forming
therefrom a composite, macro image in color of the specimen having
a field of view larger than that of the objective lens; displaying
the macro color image to the observer to provide the observer with
a larger field of view of the specimen than the observer could see
through an objective lens on the microscope; selecting a segment of
the specimen from the macro image for viewing at a higher
magnification than the low magnification; scanning and digitizing a
series of fields of view through a microscope objective lens of a
higher magnification and providing a series of digitized images;
arranging and positioning the adjacent fields of view images at
high magnification and forming therefrom a composite micro image of
the segment having a field of view larger than that of the high
magnification objective lens; and displaying to the observer both
colored composite macro image and the micro image each having a
field of view substantially larger than their respective objective
lens.
18. A method in accordance with claim 17 including the step of
marking on the macro image, the location of the micro image, so
that the observer understands where the micro image is located on
the macro image.
19. A method in accordance with claim 18 including the step of
selecting several segments each for viewing as a micro image and
marking the location of each micro image on the composite
image.
20. A method in accordance with claim 19 including the step of
recording the macro image with the marks thereon to provide a
record of the segments viewed by the observer at high magnification
to provide an audit trail for a later auditing of the segments
viewed at higher resolution.
21. A method of acquiring and analyzing biological specimens with a
computer-controlled microscope; said method comprising the steps
of: providing a biological specimen in position for scanning
through the computer-controlled microscope; scanning and digitizing
the biological specimen through the microscope at a first, low
magnification and acquiring and storing a digitized image of the
specimen in color; displaying the stored, low magnification,
digitized image in color to an observer to provide a macro image
for viewing by the observer; selecting a segment of the colored,
digitized, low magnification image for viewing at a higher
magnification than the low magnification; scanning the segment at
the higher magnification and acquiring a colored digitized image of
the segment at the higher magnification to provide a micro image of
the segment; making available in real time to the view of the
observer, the respective micro and the macro images; and performing
a quantitative analysis of the specimen from the digitized high
magnification image.
22. A method in accordance with claim 21 including the step of
marking on the macro image the position of the micro image so that
the viewer understands the location of the micro view.
23. A method in accordance with claim 20 including the steps of:
digitizing a plurality of fields of view through the microscope at
each of the low and high magnifications to form a series of
digitized image tiles; arranging the respective image tiles to form
the respective macro and micro images; displaying to the observer
the micro and macro digitized images that each have a larger field
of view than a single field of view through respective objective
lens for low and high magnification.
24. A microscopic system for analyzing a specimen and a segment of
the specimen, said apparatus comprising: a microscope having a
plurality of objective lens to acquire images at different
magnifications; a computer-controlled light illumination subsystem
on the microscope for adjusting light illumination for different
magnifications; a computer-controlled, focusing subsystem on the
microscope for adjusting the focus for different magnifications; a
computer-controlled X and Y stage to position the specimen to be
viewed at specified X and Y coordinates; an imaging subsystem
connected to a microscope to acquire and digitize the images of the
specimen; a first optical system and image acquisition system
having a first objective lens to acquire and arrange a series of
images at a low magnification to provide a macro digitized image of
the specimen having a field of view larger than the field of view
from the objective lens used for the low magnification acquisition
of images; a second optical and image acquisition system having a
second, higher magnification objective lens to acquire and arrange
a series of images at higher magnification and to provide a micro
view image of the segment having a field of view larger than the
field of view from the higher magnification objective lens; and a
display screen device to make available in real time the micro and
macro images on screens of the screen device.
25. A system in accordance with claim 24 wherein the first image
subsystem includes an X and Y position storage device for storing
the X and Y coordinates for the macro image.
26. A system in accordance with claim 24 wherein the second image
acquisition subsystem has a subsystem for reconstructing high
magnification image tiles with their X and Y coordinates positioned
to reproduce spatially high resolution image tiles into the micro
image as if the micro image was in the original image.
27. A system in accordance with claim 24 wherein the first image
acquisition subsystem has a subsystem for reconstructing low
resolution image tiles with their X and Y coordinates positioned to
reproduce spatially the low resolution tiles into a macro image of
the specimen.
28. A method for analyzing biological specimens by an image
analysis system having a computer-controlled, automated microscope
comprising the following steps: placing a biological specimen in a
microscope for viewing; using a computer terminal to control the
image analysis system and microscope to acquire low magnification,
image tiles of the specimen and to provide low magnification,
composite, macro image from the plurality of image tiles;
displaying the macro image on a screen of the computer terminal;
interactively selecting at least one point of interest on the
displayed macro, specimen image for viewing at a higher
magnification; sending signals from the computer terminal to
control the image analysis system and the computer-controlled
microscope to acquire a plurality of higher magnification, image
tiles and to provide a low magnification, composite micro image;
and analyzing the micro images of the specimen.
29. A method in accordance with claim 28 including the step of
analyzing a layer of tissue cells in the specimen for pre-invasive
neoplasia.
30. A method in accordance with claim 29 including the step of
measuring morphometric features of the specimen and measuring
texture features of the specimen from the micro images.
31. A method in accordance with claim 30 wherein the step of
interactively selecting a least one point of interest includes the
further step of selecting a series of points along a basal layer of
tissue in the specimen for analysis at high magnification.
32. A method in accordance with claim 28 wherein the steps of
interactively selecting at least one point of interest includes the
further step of selecting a plurality of points of interest on the
macro image; and recording the location of each selected point of
interest.
33. A method in accordance with claim 28 wherein the step of
interactively selecting at least one point of interest comprises
the further steps of: marking a region on the macro image on the
screen that includes portions of adjacent, low magnification image
tiles; acquiring a high magnification image tile from each of
marked low magnification tiles; and assembling these high
magnification image tiles into a micro image.
34. A method in accordance with claim 28 including the step of
simultaneously displaying on computer windows of the computer both
micro and macro images of the specimen.
35. A method in accordance with claim 28 wherein the computer
terminal is at a remote location from the computer-controlled
microscope, and including the step of first connecting the computer
terminal at the remote location over a transmission channel to the
image analysis system and the computer-controlled microscope.
Description
FIELD OF THE INVENTION
[0001] This invention relates to a method of and apparatus for
acquiring and analyzing digital images of an image viewed through a
computer-controlled automated microscope and more particularly, to
using the latter in a quantitative analysis of plant or biological
specimens.
BACKGROUND OF THE INVENTION
[0002] In the image analysis and quantification of DNA from tissue
sections as disclosed in U.S. Pat. No. 4,741,031, and also
especially in the immunohisto-chemistry assays on the kinds of cell
analysis systems disclosed in U.S. Pat. Nos. 5,086,476; 5,202,931;
and 5,252,487 issued to Bacus, there is a problem of first locating
the cancer regions for analysis under low power and then
remembering them when performing the analysis under higher power.
Specifically, the problem is that once the microscope is set up for
quantitation by image analysis under, e.g. 40.times., where all of
the diaphragms are set and light adjusted, etc., if the operator
needs to move to another tissue area, it is first desirable to
locate it at e.g. 10.times.. In fact, often regions can only be
located at this power. In order to do so however, all of the
settings (diaphragms, light levels, wavelengths of light, etc.)
have to be changed to view the tissue at this magnification.
Currently, there is no way to ensure that one could go back to the
settings at the previous 40.times. magnification and continue on
with the quantitative image analysis of that same specimen. This
necessitates finding those areas under 40.times., without changing
objectives, which is very slow and time-consuming, and often
important cancer areas can be missed.
[0003] Also, another problem with tissue analysis, at its current
state-of-the-art, is that it is not completely automated, for
example, with regard to finding structural regions such as glands,
basal layers or other important diagnostic regions. However, as set
forth in my co-pending patent application Ser. No. 701,974, filed
Aug. 23, 1996, if these regions are located, important very
sensitive diagnostic measurements can be performed, which patent
application is incorporated by reference as if fully reproduced
herein. For example, as disclosed in the aforesaid patent
application, assays are made of a variety of tissue types, both
human and animal for analysis of neoplasia in tissue, for
pre-invasive cancer in tissue, and the effects on the tissue of
chemopreventive agents. A quantitative analysis by image processing
techniques is performed on tissue types, having various
architectural features, such as breast tissue, colon tissue,
prostate tissue, esophageal tissue, skin tissue, cervix tissue,
etc. These tissues have different morphologies, and they undergo
different neoplasias usually resulting from a cellular mutation, as
may be enhanced by a carcinogen, or resulting from a cellular
proliferation rate enhanced by hormones, growth factors, or other
inducers of abnormal tissue growth. Often it is desired to quantify
small changes in the neoplasia when it is incipient or through a
series of analyses performed at close time intervals to measure
whether the neoplasia progression is increasing or has been slowed,
stopped or regressed.
[0004] Usually, the tissue specimens are cut to expose the basal
layer for review under the microscope. Typically, the quantitative
measurements are performed at 40.times. to obtain 100 to 400 tissue
images. The 40.times. objective provides a narrow field of view of
a very small portion of the entire basal layer. Often, the basal
layer is somewhat elongated and generally linear such as a basal
layer in a rat esophagus; and the analysis of the basal layer
requires examining it along its length. The basal layer in a mouse
colon is more in the form of an irregular, circular shape; and the
analysis of this basal layer requires traveling about this circular
shape. In breast tissue samples, suspected tumor areas may be at
widely-spaced points in the stained tissue; and one wants to be
able to navigate and travel to these specific suspected areas and
to do the 40.times. analysis at these areas in an efficient manner.
There is a need to allow an experienced operator to interact with
the analysis to locate and identify such regions in an interactive
manner. Especially, an interactive manner that would be familiar
and consistent with the use of a microscope manually, with high
power magnification and low power magnification simultaneously
available, but performed on a computer terminal. Such a level of
interaction is different than the interaction with the system
disclosed in the above-listed Bacus patents. There is a need to
take the level of interaction to a higher level and let each
component, the human and the computer, perform the part that it
does best, in the most cost-effective manner.
[0005] There are available on the market computer-controlled,
automated microscopes such as those sold by Carl Zeiss, Inc.,
Thornwood, N.J., under the name Axioplan 2 for taking photographic
images of a specimen in the microscopic field of view. Those
particular microscopes have computer-controlled and automatically
adjusted subsystems, such as an illumination subsystem, a focusing
subsystem, a diaphragm or optical stops subsystem, an objective
lens subsystem, or a filtering subsystem. As an operator selects
changes from one objective lens, such as one providing low
magnification, e.g., 4.times., to a higher magnification, e.g.,
40.times., the computer-automated system will turn the lens turret
to switch in the high magnification automatically and adjusts the
lens and also automatically adjusts the illumination to eliminate
glare and to provide the proper light illumination including light
density. Further, the focus is adjusted, and the proper diaphragm
openings are automatically reset. Thus, the computer-controlled,
automated subsystems automatically reset to values stored and
predetermined for each selected objective lens and the analysis
being done.
[0006] Those particular microscopes can be used to view various
objects or specimens, but are most typically used to view and to
take still photographs of biological specimens, such as tissues and
cells. Those particular microscopes lack a computer-controlled X
and Y stage for translating a specimen-carrying slide with respect
to the field of view of the selected objective lens. Currently,
pathologists and others who use such microscopes want to view the
specimen images in full color or in enhanced colors using
fluorescent illumination and/or monochromatic images using the
automated filter subsystems on the microscopes. Currently trained
pathologists or clinicians are accustomed to manually adjust and
have a microscope available to them to view larger areas of the
specimen at low magnification, and then to momentarily switch in a
new higher magnification lens to obtain a more highly magnified
image of a portion of the specimen viewed at low magnification.
Pathologists and those working in this area have created in
themselves a desire to view suspect tissue through a microscope and
appear to resist analysis systems that do not provide them this
ability.
[0007] The microscopic field of view reduces very substantially as
the magnification increases. The skill level of the clinician
and/or pathologist is important to locate viewing the most
suspicious areas or points of interest on the specimen. Sometimes,
a technician will do a first assay and analysis. A pathologist will
return to the selected points of interest or other points of
interest for review and analysis. One concern with respect to a
quantitative analysis of breast cancer tissue or prostate biopsy
tissue samples to pap smears or other tests for various cancers or
the like is that a particularly suspicious point in the tissue may
be overlooked and missed during the visual assay or for selection
for an automated review analysis. When observing at high
magnifications, the field of view is limited to very small area of
the specimen. Hence, the observer has difficulty in knowing and
remembering the actual, exact location of this small periscopic
view within the very large whole specimen.
[0008] Often, also the problem is finding or locating the tissue or
cells for view at high magnification so that artifacts and/or blank
spaces on the slide are not viewed. A number of approaches have
been proposed to prescreen and locate by an X and Y address the
cells or small points of interest from a very large number of
potential points of interest.
[0009] There are currently available commercial services for
prescreening pap smears where one can mail in slides and the
service will do a microscopic prescan at high magnification for
suspected or suspicious areas of interest which are marked and
given address locations, and also a video tape of the slide
specimen is returned by this service to the sender. The sender then
reviews the areas of interest located during the prescreening
and/or the video tape to complete the analysis.
[0010] In an attempt to locate and allow review of specified points
of interest, U.S. Pat. No. 5,428,690 to Bacus discloses a system
for prescreening of a field of cells on a specimen slide at low
magnification before the viewer. When seeing a point of interest to
be viewed at high magnification, the viewer will operate a switch
or the like to select and record the address of these selected
prescreened points of interest. Later, these prescreened points of
interest are then brought into position to be analyzed at high
magnification. This system is fairly or too slow for many uses.
[0011] A very expensive system is currently in use in which a
pathologist located at a diagnostic center is able to make a
diagnostic opinion with respect to specimens under a microscope at
a remote center. The pathologist at the diagnostic center
manipulates robotic controls to send telepathy signals via a
special, dedicated satellite or other large bandwidth channel to
control the microscope at the remote site in approximately real
time. The pathologist then can maneuver the remote microscope to
shift the microscope's field of view and to send, by telepathy, a
highly magnified, very small image back to the pathologist. This
system requires each subscriber to have a special microscope
operable by manipulation of the robotic controls at the diagnostic
center and a dedicated or large bandwidth channel to convey real
time video signals and hence results in a very high cost for the
assay being done. To assist the pathologist in staying within the
specimen at the remote site, a peripheral edge or map of the
specimen is made using a second video camera and a light box or by
using computerized scanning equipment to trace the outline or
peripheral edge of the specimen boundaries. A small circle of light
is displayed within the map of the specimen so that the pathologist
at the diagnostic center knows the location of the field of view of
the highly magnified image within the specimen. In a sense, the
pathologist is operating in real time in the same way that he would
use his own microscope at his diagnostic center except for a slight
transmission delay used to transmit the video signals of the highly
magnified image over large bandwidth channel. Although the
pathologist has a small map or peripheral outline of the specimen,
the pathologist's field of view of the actual specimen is only the
small circle of view that is coming through the microscope
objective lens. This does not help the pathologist locate
suspicious areas of interest as in a prescreening of the entire
tissue. The pathologist may switch to the lowest magnification to
get the largest field of view of a small section of the specimen,
but he never views the entire specimen at any magnification. Also,
there is no image analysis quantitative testing from the received
images at the diagnostic center; and no quantitative assaying is
done with these images at the diagnostic center.
[0012] There is a particular interest today in using the Internet
system because it is so readily accessible by users at a low cost
and using a computer and viewing screen connected to the computer.
One problem with trying to do any transmission of digitized,
microscopic, highly magnified images over the Internet is that the
bandwidth is too narrow to accommodate the tremendous amount of
stored data which needs to be transmitted. There is a need for a
system which would allow a pathologist or another person, to be
able to perform tissue analysis or quantitative assays using a
standard computer terminal from a location remote from the
automated microscope.
SUMMARY OF THE INVENTION
[0013] In accordance with the present invention, a person such as a
pathologist at a computer terminal may control an automated
microscope to acquire on a computer screen or window images of the
specimen at different magnifications as selected by the person.
Further, the person may receive on the screen a low magnification,
reconstructed image of the entire specimen to aid the person in
interactively selecting points of interest on the specimen, such as
along a basal layer of a tissue specimen.
[0014] More specifically, and in accordance with the present
invention, the microscope's small field of view limitation of a
specimen is overcome by providing to the viewer a reconstructed,
digitized and magnified image of the entire specimen (or a very
large portion of the specimen) for performing a visual analysis of
the entire tissue in full color to aid in the selection of points
of interest to be viewed at a higher magnification. This is
achieved by acquiring a large number of low magnification images of
the specimen through a microscopic scanning system, e.g. , 35 image
tiles of the specimen at 1.25.times., and then assembling and
coordinating the tiles to form an overall, low magnified image of
the specimen, i.e., a macro image of the specimen. Preferably, the
digitized macro image is reduced in size by a software system to a
smaller size, e.g., a 1/4 size image that is displayed on a local
screen or is sent over a low band width or a high band width
channel to a remote screen. Thus, the pathologist not only does not
need to have others do a slow laborious prescreening to locate
suspicious areas for analysis or for viewing at high magnification,
he can use his own experiences to go directly to the most
suspicious areas which he sees on the macro image. He can, on a
priority basis, do the most suspicious area first, followed by
lower priority areas of interest.
[0015] In accordance with the present invention, there is provided
a new and improved automated, computer-controlled microscope that
displays the low magnification composite image of the specimen to
allow the user to view and to interactively select points of
interest, each of which may be displayed at high magnification.
This is achieved by providing the user with a marker, such as a
cursor or the like, to select the defined area of interest; and to
acquire reproduced, spatially adjacent high magnification,
digitized images of the selected area of interest. More
specifically, the specimen, when it was first scanned at low
magnification to provide a macro view of the specimen, the
addresses or locations of the tile images and/or pixels for the
composite image were acquired. Therefore, any selected region of
interest in the macro image has locations to which the microscopic
stage may be automatically repositioned under a changed, higher
magnification lens to acquire higher magnification, digitized image
tiles that can be assembled into a micro image. Herein, both the
macro and micro images are formed of adjacent digitized image tiles
which have been coordinated to reproduce spatially the original
image that was scanned.
[0016] It is the high magnification images, usually at 40.times.,
that were analyzed using image processing techniques as disclosed
in the aforesaid patent application, to provide an assay or
numerical histological data for the specimen.
[0017] In accordance with the preferred embodiment of the
invention, the pathologist may select a larger region for analysis
at high resolution than can be accommodated at this magnification
on his high magnification viewing screen. He can, though, view all
of the adjacent, highly magnified, digitized image tiles on this
high magnification screen by scrolling up or down or right to left
to shift these digitized, adjacent image tiles into view on the
screen. Thus, even at a higher magnification of a region the
pathologist is able to obtain a much larger view than the small
field for the objective lens in use of adjacent tissue or cells to
give him a broader, overall perspective of what is happening or
what has happened in specific section of a specimen. For instance,
a pathologist may want to see at high magnification and to assay at
this high magnification, a suspicious area, the pathologist can
draw a mark about the area and cause it to then be assayed and
displayed.
[0018] By having displayed on a low magnification screen or split
screen of the full composite area, by having the high magnification
region being marked on the low image screen, and by having the
region being scrolled to view adjacent high magnification images on
the high magnification screen, the pathologist has available
information to guide him in the sense of helping to navigate
intelligently within the specimen in his search for cancerous
tissue or the like for further inspection or measurement of
malignant characteristics. Often, when restricted to a field of
view of an objective microscope, the pathologist has a difficult
time, in the words of a cliche, of seeing the forest for the trees.
In the present invention, the pathologist has a full, magnified,
reduced in size specimen view with the higher image area marked
thereon to guide him and to let him see the forest. He can also see
a region of the forest at higher magnification by scrolling
adjacent tree images onto the high magnification screen.
[0019] In accordance with a further aspect of the invention, the
user may elect to change to an intermediate magnification being
viewed by either switching automatically to a new objective lens
and acquiring new digitized image tiles at the intermediate
magnification or by using software to reconstruct from the existing
high and low magnification, digitized images a new intermediate
digitized image.
[0020] The preferred, low magnification image, which is reduced in
size, can be transmitted over narrow band width channels such as a
local area network or over the Internet through various servers and
computers. Likewise, the reconstructed, high magnification images
can be transmitted over such narrow band width channels. Because
the microscope is fully computer controlled, a pathologist or other
person having a split screen computer such as a PC, can be
connected to the microscope and operate it from a remote location
to obtain the macro image and to navigate to points of interest and
obtain the desire micro images. With the present invention, there
is no need for a specialized microscope at each remote location nor
for a broad band channel to send video signals in real time between
the diagnostic center and the remote location.
BRIEF DESCRIPTION OF THE DRAWINGS
[0021] FIG. 1 is a screen view of a system embodying the present
invention showing a low magnification image of a specimen on a
microscope slide in one window, a high magnification image of a
portion of the low magnification image selected by a region marker
and a control window;
[0022] FIG. 2 is a view of a display screen of the apparatus
embodying the present invention showing the control window a low
magnification window having a plurality of high magnification micro
image regions delineated therein and a high magnification window
including one or more of the micro image regions;
[0023] FIG. 3 is a view similar to FIG. 2 including the control
window but also including a low magnification region from the slide
showing regions marked by a histology grade or structure through
automatic analysis of tissue and a high magnification window
showing markings related to the grading or histology grade yielded
by the automatic analysis of tissue in combination with a window
showing a numerical score;
[0024] FIG. 4 is a block diagram of the apparatus embodying the
present invention;
[0025] FIG. 5 is a block diagram of a portion of the apparatus
shown in FIG. 4 showing details of a mechanical arrangement of a
microscope;
[0026] FIG. 6 is a flow diagram related to operation of the
apparatus;
[0027] FIG. 7 is a flow diagram of details of one of the steps in
FIG. 6;
[0028] FIG. 8 is a display screen showing control parameters to be
manipulated thereon;
[0029] FIG. 9 is a flow chart for a region outlying routine;
[0030] FIG. 10 is a flow chart for a scanning and analyzing
routine;
[0031] FIG. 11 is a schematic showing of the limits of travel of
the microscope stage with respect to the image tiles;
[0032] FIG. 11A is a perspective view of the microscope stage and
stepper motors and encoders providing a closed loop drive for the
motors;
[0033] FIG. 12 is a block diagram of a networked system allowing
multiple workstations to obtain access to diagnostic image
information and to manipulate such information locally at each
workstation;
[0034] FIG. 12A is a view of the system described in connection
with FIG. 5;
[0035] FIG. 13 is a block diagram of a remote networked system for
distributing and accessing diagnostic images and data through a
hypertext transport protocol based server directly or over a packet
network;
[0036] FIG. 14 is a view of a low magnification, reconstructed
image from a basal layer of rat esophagus;
[0037] FIG. 14A is a view of a high magnification, reconstructed
image from a selected point of interest from FIG. 14;
[0038] FIG. 15 is a view of a low magnification image of a mouse
colon having a basal layer;
[0039] FIG. 15A is a view of a reconstructed macro image of a mouse
colon;
[0040] FIG. 16 is a schematic view of an analysis from regions of a
basal layer;
[0041] FIG. 16A is a schematic view of an analysis to provide to a
Z score; and
[0042] FIG. 17 is a schematic view showing texture analysis tests
for regions.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0043] Referring now to the drawings, and especially to FIGS. 4 and
5, apparatus for synthesizing low magnification and high
magnification microscopic images is shown therein and generally
identified by reference numeral 10. The system includes a computer
12 which is a dual Pentium Pro personal computer in combination
with a Hitachi HV-C20 video camera 14 associated with a Zeiss
Axioplan 2 microscope 16. The computer system 12 is able to receive
signals from the camera 14 which captures light from the microscope
16 having a microscope slide 18 positioned on an LUDL encoded
motorized stage 20. The encoded motorized stage 20 includes a MAC
2000 stage controller for controlling the stage in response to the
computer 12. A microscope slide 18 includes a biological specimen
21 which is to be viewed by the microscope and whose image is to be
digitized both at low magnification and at high magnification as
selected by a user. The low magnification digitized image is then
displayed on a 21 inch Iiyama video display monitor 22 having
resolution of 1600 by 1200 to provide display screens of the type
shown in FIGS. 1 through 3 including a low magnification image 24,
for instance, at 1.25 power, a high magnification image 26, for
instance at 40 power and a control window or image 28. The low
magnification image may have identified therein a region 30 which
is reproduced at high magnification in high magnification screen or
window 26 so that a pathologist or other operator of the system can
review architectural regions of interest in low magnification image
24 and simultaneously view them in high magnification in the high
magnification screen or window 26 to determine whether the cells
forming a portion of the architectural feature need be examined
further for cancer or the like or not.
[0044] The computer 10 is constructed around a PCI system bus 40
and has a first Pentium Pro microprocessor 42 and a second pentium
pro microprocessor 44 connected thereto. The system bus 40 has
connected to it a PCI bus 50 and an ISA bus 52. The PCI bus 50 has
a SCSI controller 60 connected thereto to send and receive
information from a hard disk 62. The hard disk 62 also is coupled
in daisy chain SCSI fashion to a high capacity removal disk and to
a CD Rom drive 66. The hard disks 62 contains the programs for
operating the system for controlling the microscope 16 and for
processing the images as well as for doing a quantitative analysis
of the selected portions of the histological specimens being viewed
on the slide 18. The system bus 40 also has connected to it a
random access memory 70 within which portions of the program being
executed are stored as well as a read only memory 72 for holding a
bootstrap loader as well as portions of the basic input/output
operating system. A floppy disk controller 74 is coupled to the
system bus 40 and has connected to it a floppy disk drive 76 for
reading and writing information to a floppy disk as appropriate. A
mouse controller 80 is coupled to the system bus and has a mouse 82
which operates as a pointing device for controlling manipulations
on the screen 22 and within the windows 24, 26 and 28. A keyboard
controller 90 is connected to the system bus and has a keyboard 92
connected thereto. The keyboard 92 may be used to send and receive
alpha numeric signals to other portions of the computer. An audio
controller 100 has a plurality of speakers 102 and a microphone 104
connected thereto for audio input and output and is coupled to the
system bus 40. A network interface, such as a network interface
card 104, is connected to the system bus and can provide signals
via a channel 106 to other portions of a network or internet to
which the system may be connected. Likewise, signals can be sent
out of the system through a modem 110 connected to the ISA bus 52
and may be sent via a channel 112, for instance, to the internet. A
printer 116 is connected via a parallel I/O controller 118 to the
system bus in order to provide printouts as appropriate of screens
and other information as it is generated. A serial I/O controller
122 is connected to the system bus and has connected to it a camera
controller 124 which is coupled to CCD sensors 126 in the cameras.
The CCD sensors 126 supply pixel or image signals representative of
what is found on the slide 18 to an Epix pixci image acquisition
controller 130 coupled to the PCI bus 50.
[0045] The microscope 16 includes a base 140 having a stage 20
positioned thereon as well as an objective turret 142 having a
plurality of objectives 144, 146 and 148 thereon. The objective
144, for instance, may be of 1.25.times. objective. The objective
146 may be a 20.times. objective. The objective 148 may be a
40.times. objective. Signals from the camera sensors and controller
are supplied over a bus 128 to the image acquisition system where
they are digitized and supplied to the PCI bus for storage in RAM
or for backing storage or the hard disk 62.
[0046] When a specimen is on the slide 18 the stage 20 may be
manipulated under the control of the computer through a stage
controller 160 coupled to the serial I/O controller 122. Likewise,
a microscope controller 162 controls aspects of the microscope such
as the illumination, the color temperature or spectral output of a
lamp 168 and the like. For instance, in normal operation, when a
specimen is placed on the slide, specimen slide 18 is placed on the
stage 20 in a step 200, as shown in FIG. 6, the processors 42 or 44
send a command through the system bus to cause the serial I/O
controller 122 to signal the microscope controller to change
magnification to 1.25.times. in a step 202. This is done by
rotating the objective turret of the Axioplan 2 microscope to
select the objective 144. Likewise, the controller sets the color
temperature of the lamp 168, sets a pair of neutral density filter
wheels 170 and 172 and sets a field diaphragm 174 for the correct
illumination. A condenser diaphragm 176 is also controlled and a
color filter wheel 180 may also be controlled to apply the
appropriate filter color to the CCD censors 126 in the camera. The
entire slide is then scanned in a step 204. The images are tiled
and melded together into the overall image 24 supplied on the
screen 22 to provide the operator in the step 206 with a visually
inspectable macro image of relevant regions of the slide of
interest.
[0047] In order to provide the magnified image, the mouse may be
moved to identify a marker segment or region which, for instance,
may be a rectangular region (as shown as 30 in FIG. 1) which will
cause the microscope to change magnification as at step 208 to
4.times., 20.times., 40.times., etc., by rotating the turret to
bring the appropriate objective lens system into viewing
position.
[0048] Next the user, in a step 209a, uses the mouse to select the
region on the macro image in order to select the micro image to be
viewed on the screen 22. In a step 209b a test is made to determine
whether the user has commanded continued inspection. If the user
has, a test is made in a step 209c to determine if the
magnification is to be changed by changing the selected objective.
In the event the magnification is to be changed control is
transferred to the step 208. If the magnification is to remain
unchanged control is transferred to the step 209a. In the event
inspection is not to continue the region selected is outlined for
higher magnification scan in a step 209d. In a step 209e, a command
may be received to scan or acquire the higher magnification image
for display in screen 26. The image may then be archived for later
analysis, displayed or analyzed immediately.
[0049] In order to perform the magnification called for in step
206, the overall illumination and control of the microscope will be
controlled so that in a step 210 the objective turret 142 will be
rotated to place the higher power objective above the slide 18. In
a step 212 voltage to the lamp will be changed to adjust the lamp
168 to provide the proper illumination and color temperature as
predetermined for the selected objective. In a step 214, the
condenser diaphragm 176 will have its opening selected as
appropriate to provide the proper illumination for that objective.
In a step 216, the filter turret 180 will select the proper light
wavelength filter to be supplied to the camera sensors. For
instance, a red, blue or green filter, as appropriate, particularly
if the specimen has been stained. In a step 218 the field diaphragm
174 will have its opening changed. In a step 220 the neutral
density filter wheel 170 will select a neutral density filter and
in a step 222 the neutral density filter wheel 172 will also select
a neutral density filter. In a step 224 the X, Y and Z offsets will
be used for reconstruction of the recorded image at the
magnification and in a step 226 the current position will be read
from encoders in the stage which are accurate to 0.10 micron.
[0050] In order to identify the selected region the mouse is moved
to that area of the region in a pointing operation in a step 240 as
shown in FIG. 9. The mouse may be moved to draw a box around the
region selected. In a step 242 the X and Y screen points are
computed for the edges of the regions selected and the computed
image or pixel points are translated to stage coordinate points in
order to control the stage of the microscope. In a step 244 a list
of all of the X fields for positioning the stage for the objective
is stored in random access memory and may be backed up on the hard
disk. The information from the X offsets for the objective and the
stage offsets is used as well as the size of the field to position
the slide properly under the objective to capture the micro
image.
[0051] When the slide has been positioned properly, as shown in
FIG. 10 in a step 250 the stage is positioned for each of the X and
Y coordinate values in stage coordinate values and the digitized
image is captured by the cameras and stored in RAM and backed up on
the hard disk. The image may be then analyzed quantitatively in
various manners such as those set forth in the
previously-identified United States application. Optionally the
image may be stored for archival purpose in a step 254.
[0052] In order to override the specific control functions that
take place as shown in FIG. 7, a screen is provided as shown in
FIG. 8 wherein the XY step size can be edited, the X, Y and Z
offset can be edited, the lamp voltage can be selected, the neutral
density filter can be selected as well as the opening of the field
diaphragm and several other microscopic characteristics. FIG. 8 is
a view of the settings of the microscope objective properties of
the Axioplan 2, computer-controlled microscope.
[0053] The X and Y positioning is specifically carried out as shown
in FIG. 11 where the slide 18 is shown with a slide boundary 270,
272, 274 and 276. Stage boundary for limits of the stage travel for
purposes of the stage the stage can be moved all the way from an
upper left hand corner of travel 276 to a lower right hand corner
of travel 280. At the upper left hand bounded corner of travel 278
limits which a signal that the end of travel has been reached and
the stage is then translated a short distance 282 in the extra
action and a short distance 284 in the Y direction to define the
first tile 288 in terms of a reference point 290 at its upper left
hand corner. Since the size of the macro image tile 288 is known,
the next macro image tile 292 may be placed contiguous with it by
moving the stage appropriately and by measuring the location of the
stage from the stage in counters without the necessity of
performing any image manipulation. The image tiles 288 and 292 may
be abutted without any substantial overlap or they may be
overlapped slightly, such as a one pixel with overlap, which is
negligible insofar as blurring of any adjacent edges of abutted
image tiles. The upper left hand corner 300 of the tile 292 defines
the rest of 292 and other tiles can be so defined. Micro image
tiles can likewise be defined so that they are contiguous but not
substantially overlapping, as would interfere with the composite
image. This avoids the problems encountered with having to perform
extended computations on digital images in a frame storer or
multiple frame storage in order to match or bring the images into
contiguity without blurriness at the edges of contiguous image
tiles. It may be appreciated as shown in FIG. 2 that the low power
image 24 has a plurality of micro images defined therein which are
tiled and which are shown in higher magnification as individual
tiles 312, 314, 316 and the like in FIG. 2.
[0054] In addition, the region 310 when magnified as shown in the
window 26 may exceed the bounds of the window and thus the window
may include scroll bars or other means for allowing the image 310
which is larger than the window 26 to be examined from within the
window 26.
[0055] The stage 200 is best seen in FIG. 11A and includes the X
and Y stepper motors 279 and 281 with their respective encoders,
which provide a closed loop system to give the 0.1 micron accuracy
versus the usual 5 or 6 micron accuracy of most microscope stages
without a closed loop system. This closed loop system and this very
high accuracy allow the abutting of the tile images for both high
magnification and low magnification images without the substantial
overlap and the time-consuming and expensive software currently
used to eliminate the overlap and blurriness at the overlapping
edges of adjacent image tiles. With the precisely positioned stage
and by using the tiling system described in connection with FIG.
11, where the slide is precisely positioned relative to a center
point CP for the slide, and the known position of point 278 is
always taken from the same point, the tiles may be positioned
precisely in a horizontal row and precisely in vertical rows to
reconstruct the macro image and the micro image. This
reconstruction is done without the use, as in the prior art, of
extensive software manipulation to eliminate overlapping image
tiles, horizontally or vertically or the haphazard orientation of
image tiles.
[0056] Furthermore, as shown in FIG. 3, the low power window 24,
high power window 26 and control window 28 can be used in
conjunction with reporting of quantitative analysis data,
histograms, etc. for the specimen being viewed; and such analysis
information may be provided as a visual output in a window 320.
Each of the various regions 30 that a pathologist may follow in
marking various features in the low power window 24 and the high
power window 26 may be reflected in both windows in order that an
audit trail is provided for the system.
[0057] The present invention also includes the facility for
allowing remote diagnostics to occur by being able to couple the
system either over a network communication facility to an intranet,
for instance via the network interface, or via a modem or other
suitable connection, to an internet so that once the image has been
scanned and stored in memory on hard disks or other storage, remote
users may be able to access the low magnification image as well as
the high magnification image and move around within both images to
make determinations as to the histological characteristics of the
samples via Z scores.
[0058] An additional feature of the system includes a plurality of
networked workstations coupled to a first computer console 12
having a display screen 22 connected to the microscope 14.
Satellite work stations 350 and 352 are substantially identical to
the work station 12 including respective computers 354 and 356
coupled to displays 358 and 360. The devices can be manipulated
through input devices 360 and 362 which may include a keyboard,
mouse and the like. Also a third device can be connected including
a work station 370, having a display 372, a computer 374 and an
input device 376. Each of the devices is connected over respective
network lines 380, 382, 384 to the computer 12 which transmission
may be via either net or the like. Each of the different operators
at the physically separate viewing stations can locate regions from
the view of entire tissue cross sections via a macro view and label
the regions for subsequent scanning and/or quantitative analysis. A
single operator at the instrument station 12 can locate regions to
view the entire tissue cross section. Those regions can be labeled
for subsequent scanning and/or quantitative analysis with
subsequent review and physically remote viewing stations, for
instance, in an operating room or in individual pathologists'
signout areas in order to review analysis results while still
maintaining and reviewing the entire macro view of the tissue
and/or the individual stored images from which the quantitative
results were obtained. The viewing stations 350, 352 and 370 can
comprise desk top computers, laptops, etc. There is no need for a
microscope at the network stations 350, 352 and 370.
[0059] In a still further alternative embodiment, remote
workstations 400, 402, 404, 406 and 408 may be connected through a
server 410 which may be supplied via a packet switched network. The
server 410 and may be a hypertext transport protocol based server
of the type used for the World Wide Web or may be a telnet type
server as used previously in internet remote operation
applications. The server 410 communicates via a communications
channel 414 with a local computer 416 having a display 418
associated therewith, the local computer 416 being connected to the
microscope 420. Each of the remote work stations 400, 402, 404, 406
and 408 may perform the same operations as the stations 350, 352
and 370 although they do it from nearby buildings or even from
around the world, thus providing additional flexibility for others
to make use of the specimen obtained and being viewed under the
microscope 420. In addition, stored images may be disseminated
through the server 410 to the remote servers 400 through 408 for
further analysis and review.
[0060] In FIG. 14, there is illustrated on screen 28 a basal layer
431a of a cut cross-section of a rat esophagus. The basal layer is
elongated and linear in a downward direction, and the selected
point of interest is shown as a box 30 on the basal layer on the
composite, low magnification image. The high magnification image 26
of this selected point of interest is shown on screen 26 in FIG.
14A. In FIG. 15 is shown a mouse colon as a reconstructed, low
magnification macro image 28 which has been reduced {fraction
(1/16)}th in size. The macro image 26 is shown in FIG. 15A, and the
marking therefore is shown in FIG. 15.
[0061] The analysis for texture and for morphological features used
to analyze a series of regions 30 on the elongated basal layer that
were analyzed at high magnification are shown in FIGS. 16, 16A and
17. The manner of doing these tests and of obtaining a Z score or
grade is disclosed in the aforesaid patent application.
[0062] While there has been illustrated and described a particular
embodiment of the present invention, it will be appreciated that
numerous changes and modifications will occur to those skilled in
the art, and it is intended in the appended claims to cover all
those changes and modifications which followed in the true spirit
and scope of the present invention.
* * * * *