U.S. patent number 3,864,564 [Application Number 05/400,915] was granted by the patent office on 1975-02-04 for acquisition system for slide analysis.
This patent grant is currently assigned to Corning Glass Works. Invention is credited to William J. Adkins.
United States Patent |
3,864,564 |
Adkins |
February 4, 1975 |
ACQUISITION SYSTEM FOR SLIDE ANALYSIS
Abstract
A closed loop scanning and positioning system for finding and
positioning white blood cells includes a multifaceted rotating
mirror, an optical system, two photoelectric sensors, two light
sources and a logic circuit. The logic circuit produces signals
which are used to drive a microscope stage in x and y directions to
cause the blood cell to be positioned within a small aperture.
Inventors: |
Adkins; William J. (Raleigh,
NC) |
Assignee: |
Corning Glass Works (Corning,
NY)
|
Family
ID: |
23585526 |
Appl.
No.: |
05/400,915 |
Filed: |
September 26, 1973 |
Current U.S.
Class: |
250/548; 318/577;
359/393; 250/234; 318/640 |
Current CPC
Class: |
G01N
15/1468 (20130101); G01N 21/5911 (20130101); G06K
9/00127 (20130101); G06K 9/20 (20130101) |
Current International
Class: |
G06K
9/00 (20060101); G01N 21/59 (20060101); G01N
15/14 (20060101); G06K 9/20 (20060101); G01j
001/20 () |
Field of
Search: |
;250/201,222PC
;356/39,40 ;235/92PC ;318/565,577 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Lawrence; James W.
Assistant Examiner: Nelms; D. C.
Attorney, Agent or Firm: Zebrowski; Walter S. Kurtz; Richard
E.
Claims
What is claimed is:
1. In a system producing an electrical output representing the
optical characteristics of an analytical slide containing a
specimen of interest, including:
conversion means producing said electrical output representing the
optical characteristics of said slide,
means for projecting an optical image of said analytical slide to
said conversion means,
a light sensing device,
means for successively scanning a portion of said optical image,
said light sensing device producing an output representing the
light absorption of the area of the slide being scanned,
a pulse shaping means connected to the output of said light sensing
device and producing an acquisition signal when the area of the
slide being scanned contains a specimen of interest, and
a positioning mechanism for positioning said slide, an acquisition
system comprising:
synchronizing means producing a synchronizing signal at a
predetermined point in each scan, and
logic circuitry responsive to said synchronizing signal and to said
acquisition signal for producing control signals, said control
signals being used to control said positioning mechanism to
position said slide so that the image of a specimen of interest is
centered on said conversion means.
2. The system recited in claim 1 wherein said synchronizing means
comprises:
a rotating mirror,
a light source focused on said rotating mirror, and
a second light sensing device, said second light sensing device and
said light source being positioned so that reflected light striking
said second light sensing device is in time relation to the
beginning of a scan of said mirror across said slide, said second
light sensing device producing a synchronizing signal upon
reception of said light.
3. The system recited in claim 1 wherein said logic circuitry
includes means for producing a normal stage motion signal if no
specimen is detected during a scan said normal stage motion signal
being applied to said positioning mechanism to move said slide by
an increment in a direction orthogonal to the direction of
scan.
4. The system recited in claim 1 wherein said logic circuitry
includes:
means responsive to said synchronizing signal for producing a
center pulse occurring approximately in the middle of each scan,
and
means responsive to the occurrence of an acquisition signal before
or after said center pulse for producing directional signals
representing the direction of motion of said slide along the
direction of scanning required to center said specimen on an
aperture of said conversion means, said directional signals being
applied to control said mechanism.
5. The system recited in claim 4 wherein said logic circuitry
further comprises:
means for producing a specimen-centered signal when said
acquisition pulse coincides with said center pulse thereby
indicating that said specimen is centered on said aperture.
6. The system recited in claim 4 wherein said means for producing a
center pulse comprises:
an oscillator producing clock pulses, a pulse counter, said clock
pulses being applied to said pulse counter upon the occurrence of
said synchronizing pulse, and
a decoder, the outputs of said counter being applied to said
decoder, said decoder producing said center pulse upon the
occurrence of a predetermined count in said counter.
7. The system recited in claim 4 wherein said means for producing
directional signals comprises:
a shift register having a first stage which is set by said
synchronizing signal and which is reset by said center pulse and a
second stage which is switched by said acquisition signal, said
first stage being connected to said second stage so that the state
of said first stage is set into said second stage when said
acquisition signal occurs, the output of said second stage being a
bistable directional signal which indicates whether said
acquisition signal occured before or after said center pulse.
8. The system recited in claim 6 further comprising:
a second decoder, the outputs of said counter being applied to said
second decoder to produce an end pulse coincident with the end of
each scan,
a specimen-centered flip-flop which is set by the coincidence of
said acquisition signal and said center pulse, and
a gate, said end pulse and the output of said specimen-centered
flip-flop being applied to said gate, said gate producing a
specimen-centered signal which is applied to said conversion means
at the end of a scan so that said conversion means converts the
optical image of a specimen which is centered on said aperture into
an electrical output.
9. The system recited in claim 8 further comprising:
an acquisition flip-flop, said acquisition flip-flop being set by
said acquisition signal, and
a second gate, said end pulse and the output of said acquisition
flip-flop being applied to said second gate to produce a
directional signal, said directional signal being applied to said
positioning mechanism to move said slide along the direction of
scanning required to center said specimen on said aperture.
10. The system recited in claim 9 further comprising:
means for producing a normal stage motion signal if no specimen is
detected during a scan, said normal stage motion signal being
applied to said positioning mechanism to move said slide by an
increment in a direction orthogonal to the direction of scan,
and
priority circuitry connected to enable said logic circuitry to
produce only one of said specimen-centered, directional and normal
stage motion signals to the exclusion of others in the order of
priority of, a specimen-centered signal, a directional signal, and
a normal stage motion signal.
11. The system recited in claim 6 wherein said oscillator has an
adjustable frequency to adjust the length of each scan.
12. The system recited in claim 6 comprising:
an adjustable one-shot multivibrator, said synchronizing pulse
being applied to trigger said one-shot multivibrator, the output of
said one-shot multivibrator being applied to said oscillator to
start the production of said clock pulses, said one-shot
multivibrator having an adjustable monostable time period so that
said center pulse occurs at a position in said scan which coincides
with the center of the field of view of said conversion means.
13. The system recited in claim 1 wherein said means for projecting
includes a beam splitter which projects one slide image to said
conversion means and another slide image to said rotating
mirror.
14. The system recited in claim 1 wherein said conversion means
includes an aperture, said positioning mechanism being controlled
to center the image of a specimen of interest on said aperture.
15. The system recited in claim 4 wherein said synchronizing means
comprises:
a rotating mirror,
a light source focused on said rotating mirror, and
a second light sensing device, said second light sensing device and
said light source being positioned so that reflected light striking
said second light sensing device is in time relation to the
beginning of a scan of said mirror across said slide, said second
light sensing device producing a synchronizing signal upon
reception of said light.
Description
BACKGROUND OF THE INVENTION
This invention relates to a system for converting the optical image
of a laboratory slide to electrical signals and more particularly
to an acquisition system for bringing a specimen of interest on the
slide into position for analysis.
In the analysis of blood samples, the blood is smeared on a
laboratory slide and the smear is stained. By counting the
leukocytes on the stained smear, laboratory technicians perform
what is referred to as a white blood cell differential. Automation
of this differential has significant economic impact because the
differential is performed so frequently at every hospital. A thesis
by J. W. Backus, "An Automated Classification of the Peripheral
Blood Leukocytes by Means of Digital Image Processing", University
of Illinois, Chicago, 1971, describes one automated system.
In a system developed by my co-employees, a scanning unit (in this
case a T.V. camera) linearly sweeps a vidicon target subjected to
intense illumination which passes through the smeared slide. Such a
system is described in copending application Ser. No. 353,004,
filed Apr. 20, 1973.
In order to count and classify the blood cells on a slide it is
necessary to successively find each blood cell and focus its image
on the vidicon target. The focusing system is described in
copending application Ser. No. 399,619, filed Sept. 21, 1973, Amos
et al. The system for successively locating and bringing each blood
cell into a position for analysis is the subject of the present
application.
SUMMARY OF THE INVENTION
In accordance with this invention a specimen of interest on an
optical slide is positioned with respect to a viewing aperture by a
logic circuit which receives signals from light sensing devices. A
rotating mirror successively scans portions of the optical image of
the slide across one of the light sensing devices. The logic
circuit produces control signals which are used in positioning of
the slide. If no specimen of interest, in this case a blood cell,
is detected during a scan, the logic produces a signal which moves
the microscope slide in the y direction, orthogonal to the
direction of scanning so that another field of view can be
scanned.
A second light sensing device receives light at the beginning of
each scan of the rotating mirror. This light sensing device
produces a synchronizing pulse which is applied to the logic
circuit. If a cell is detected during a scan, the logic circuit
compares the time occurrence of the acquisition of the cell to the
time of the synchronizing pulse. In this manner the logic circuit
produces signals which specify the direction in which the slide is
to be moved along the x axis in order to center the cell in the
viewing aperture.
The foregoing and other objects, features and advantages of the
invention will be better understood from the following more
detailed description and appended claims.
DESCRIPTION OF THE DRAWINGS
FIG. 1 is an optical and electrical schematic diagram of the system
of this invention;
FIGS. 2A-2D depict the acquisition of a blood cell; and
FIGS. 3A-3C show the electrical schematic of the logic circuit.
DESCRIPTION OF THE PREFERRED EMBODIMENT
FIG. 1 shows the optics for a system for scanning and counting
leukocytes on a blood smeared slide 10. Light from the lamp 12
passes through lens 14, is reflected from fold mirror 16, and
passes through condenser lens 18. The light passes through the
slide 10 and is collected by the objective lens 20. A first beam
splitter 22 reflects approximaterly 40 percent of the light to the
automatic focus and acquisition system of which this invention is a
part. The remainder of light is reflected by mirrors 24 and 26 to
an optical-to-electrical convertor which scans and analyzes a blood
cell whose image is centered on the aperture 28. The light image
passing through the aperture 28 is applied to relay lens 30, to
optical preprocessor 32 and to the compensator 34. The cell image
is applied to vidicon type T.V. camera 36 which produces electrical
signals which are ultimately converted into digital signals
representing the characteristics of each blood cell. This
conversion is more fully described in the aforementioned Cotter
application.
A portion of the focused light from beam splitter 22 strikes a
rotating multifaceted mirror 38. Light striking mirror 38 forms an
image of the slide which is parfocal with the image applied to the
converter. The rotating mirror reflects a portion of the slide
image onto a light sensing device 40. Light sensing devices 42 and
44 are used in the focusing system as more fully described in the
copending Amos et al application. A very narrow slice of the slide
image is scanned as the mirror 38 rotates. The light sensing device
40 produces an output proportional to the light absorption of the
area of the slide being scanned. This signal is amplified and peak
detected in the pulse shaping circuits 46. This circuit produces
acquisition pulses indicating the detection of a blood cell on a
slide only for signals above a given threshold.
A second light source 48 is focused on the rotating mirror 38 and
the focused light is swept across a second light sensing device
such as photocell 50. The light sensing device 50 is adjusted so
that the light from source 48 strikes it at a predetermined
position in the scan. In this case, the light strikes photocell 50
in time relation to the beginning of a scan of the mirror across
the slide. The photocell 50 produces a synchronizing signal to mark
the beginning of the sweep by a given mirror facet.
The present invention is directed to the system which centers an
image of each blood cell on the slide onto the aperture 28. The
acquisition pulses and synchronizing pulses are applied to logic
circuitry 52. The logic circuitry utilizes these pulses to
determine if a specimen of interest is in the field of view of one
of the scanning segments. If so, the circuitry determines which way
the stage must move in order to center the specimen in the
aperture. As each facet of the rotating mirror sweeps the field,
one of four signals is generated by the logic circuitry. If no
specimen is encountered, a normal stage motion signal is generated.
This causes the y positioning motor 54 to move the slide one step
in the y direction so that another field of view can be scanned.
This one step per mirror facet motion continues until a specimen is
encountered by the acquisition sensor 40. At this time a
directional signal is produced. The directional signal indicates
whether the specimen is to be moved to the right or left, .+-. x
direction, in order to center the cell on the aperture. The x
positioning motor 56 moves the slide in either the +x or the -x
direction. This motion continues until the logic circuitry produces
a cell-centered signal. At this time all slide motion is halted,
the cell specimen is centered in the aperture 28, and the vidicon
36 is enabled to allow it to make an analysis of the cell. The
stage motors 54 and 56 are stepping motors which move the stage a
predetermined distance for each applied pulse.
In the overall system of which this invention is a part, a small
digital computer is used in the analysis and storage of information
regarding the blood cells. It is convenient to use this digital
computer for control purposes. Therefore, it will be preferred to
apply the normal stage motion, directional and cell-centered
signals from the logic circuitry to the digital computer. However,
for purposes of explanation it is clearer to describe the logic
circuitry as controlling the x and y positioning motors 54 and 56
and the vidicon 36. The modifications required to use the digital
computer for control are well understood by those skilled in using
these devices.
The operation can best be explained with reference to FIGS. 2A-2D.
These figures depict the image of a blood smeared slide. Included
in this image is a specimen of interest, notably the blood cell 58.
The figures depict the image in relation to the aperture 28 and the
figures depict the image in relation to the field of view which is
scanned and which is shown as the shaded areas. (It should be
remembered that the image applied to aperture 28 is parfocal with
the image scanned by the rotating mirror 38. Therefore, it is
accurate to depict the scan field of view as the shaded area in
relation to the aperture 28 in this manner.)
A first scan by one facet of the mirror 38 is depicted in FIG. 2A.
During this scan no blood cell is detected by the acquisition
detector 40. Therefore, the logic circuitry produces a normal stage
motion signal which moves the slide by one increment in the y
direction. During the next scan depicted in FIG. 2B, the
acquisition detector 40 does not produce an acquisition pulse.
Again a normal stage motion signal moves the slide one increment in
the y direction to the position depicted in FIG. 2C. During this
scan the acquisition detector 40 produces a pulse indicating that a
blood cell lies in the field of view of this scan. Further, the
logic circuitry detects that the cell lies to the left of the
aperture 28. Therefore, the logic circuitry produces a directional
signal which moves the slide in the +x direction. This directional
signal is produced until the slide is positioned with the cell 58
centered on the aperture 28 as depicted in FIG. 2D.
The logic circuitry for producing signals in this manner is shown
in FIGS. 3A-3C. A synchronizing pulse from the synchronizing
detector 50 is applied to set the flip-flop 60 at the beginning of
each scan by a mirror facet. If there are more than one sync pulse
during a scan, due to noise, only the first sync pulse sets the
flip-flop 60. The flip-flop 60 triggers the one-shot multivibrator
62. By adjusting the monostable time period of multivibrator 62, it
is possible to center the aperture with respect to the viewing
binoculars used by the operator.
One-shot multivibrator 62 triggers the one-shot multivibrator 64
which produces a pulse of predetermined width. This is used to set
the flip-flop 66. When the flip-flop 66 is set an oscillator 68 is
turned on. This triggers clock pulses which are applied to one-shot
multivibrator 70 which produces pulses of a predetermined width.
These pulses are applied to the binary counters 72 and 74. The
outputs of the counters 72 and 74 are decoded by the decoding gates
76 and 78. The decoding circuitry divides each scan line into 60
equal increments. A center pulse is produced at the middle of the
scan and an end pulse is produced at the end of the scan. In order
to produce the center pulse, the decoding gate 76 has inputs from
counters 72 and 74 such that the output of gate 76 goes up when the
counters receive the twenty-ninth clock pulse. The thirtieth clock
pulse passes through the gate 80. This is the center pulse.
The decoding gate 78 has inputs from counters 72 and 74 such that
the output of gate 78 goes up on the fifty-ninth clock pulse. The
sixtieth clock pulse passes through the gate 82. This is the end
pulse. It resets flip-flops 60 and 66. This cuts off the
oscillator. The end pulse also provides a reset for the counters 72
and 74.
If an acquisition pulse occurs during a scan, it passes through AND
gate 84 and inverter 86 to set the acquisition flip-flop 88. If an
acquisition pulse occurs at a time which coincides with the center
pulse, the center pulse passes through AND gate 94 which is enabled
by the acquisition pulse from gate 84. The output of gate 94 passes
through inverter 96 to set the cell-centered flip-flop 98.
Flip-flop 98 enables the AND gate 100 and the end pulse passes
through it to produce the cell-centered pulse.
If an acquisition pulse occurs during a scan, but it does not
coincide with the center pulse, a directional signal is produced
which indicates the direction of movement along the scan required
to center the acquisition pulse. Whether the acquisition pulse
occurs before or after the center pulse is detected by a shift
register which includes a first stage flip-flop 102 and a second
stage flip-flop 104. The first stage flip-flop is initially set; it
is reset by the center pulse. Upon the occurrence of the
acquisition pulse, the state of the first stage flip-flop 102 is
set into the second stage flip-flop 104. If the acquisition pulse
occurs before the center pulse, a set state is transferred from the
flip-flop 102 to flip-flop 104. If the acquisition pulse occurs
after the center pulse, a reset condition is transferred from
flip-flop 102 to flip-flop 104. If an acquisition has occurred, the
end pulse will pass through AND gate 106 and interrogate the AND
gates 108 and 110. If the acquisition occurs before the center
pulse, the flip-flop 104 is set so that the connection from
flip-flop 104 to the gate 110 is high. This allows the pulse from
gate 106 to pass through gate 110 to produce a directional signal
indicating a move in the +x direction. Conversely, if the
acquisition pulse occurs after the center pulse, the flip-flop 104
is reset and the inverter 112 applies a high condition to the gate
108. The pulse from gate 106 passes through the gate 108 to produce
a directional signal indicating a move in the -x direction. If
there is no acquisition pulse during a scan, the acquisition and
cell-centered flip-flops 88 and 98 will still be reset at the time
of the end pulse. The bottom outputs of these flip-flpos acts
through OR gate 90 to enable the AND gate 92. The AND gate 92
passes the end pulse to produce a normal stage motion signal.
Priority circuitry is connected so that only one of the
cell-centered, directional, and normal stage motion signals can be
produced to the exclusion of the others. With the circuitry
described thus far, it would be conceivable that all three signals
would be produced at the end of a given scan. Of course it is
necessary that only one produced so as to avoid confusion. The
highest priority signal is the cell-centered signal. When the
flip-flop 98 is set, the lower output from this flip-flop is low.
This acts through the gate 114 to hold the acquisition flip-flop 88
in the reset condition. This prevents an end pulse from passing
through AND gate 106 to produce a directional signal. The bottom
output of flip-flop 98, which is low because the flip-flop is set,
acts through OR gate 90, to block AND gate 92. This prevents the
production of a normal stage motion signal.
The second priority is the production of directional signals. If
the acquisition flip-flop 88 is set, the lower output of the
flip-flop is applied through OR gate 90 to block the gate 92. No
normal stage motion pulse can be produced.
The only time a normal stage motion pulse is produced is at the end
of a scan in which neither the acquisition flip-flop 88 nor the
cell-centered flip-flop 98 has been set. In this case, the end
pulse can pass through both gates 90 and 92 and produce a normal
stage motion pulse.
Refer once more to the adjustment for the one-shot multivibrator
62. This manual adjustment moves the center pulse in relation to
the sync pulse. It is desirable that the center pulse occur with
respect to the scan at a point which is approximately in the middle
of the field of view of the camera 36. The one-shot multivibrator
62 can be adjusted until this condition is met. Similarly, the
frequency adjustment of the oscillator 68 can be adjusted. By
lowering the frequency of these pulses, the sixty clock pulses will
encompass a greater length of scan. Similarly, increasing the
frequency decreases the length of scan.
While a particular embodiment of the invention has been shown and
described, various modifications are within the true spirit and
scope of this invention. The appended claims are, therefore,
intended to cover all such modifications.
* * * * *