U.S. patent application number 11/403832 was filed with the patent office on 2006-08-31 for confocal microscope apparatus.
This patent application is currently assigned to TOKAI UNIVERSITY EDUCATIONAL SYSTEM. Invention is credited to Hideyuki Ishida, Takeo Tanaami.
Application Number | 20060192075 11/403832 |
Document ID | / |
Family ID | 33549133 |
Filed Date | 2006-08-31 |
United States Patent
Application |
20060192075 |
Kind Code |
A1 |
Ishida; Hideyuki ; et
al. |
August 31, 2006 |
Confocal microscope apparatus
Abstract
A confocal microscope apparatus has a confocal scanner for
scanning a sample with shifting a focal position of a light beam in
a direction perpendicular to an optical axis, a moving mechanism
for moving the focal position of the light beam in an optical axis
direction, a camera for picking up an image of the sample with the
light beam, and a movement control unit for controlling the moving
mechanism to move the focal position of the light beam by a
predetermined distance in the optical axis direction for every
vertical synchronizing signal of the camera in synchronization with
the vertical synchronizing signal. A high-speed three-dimensional
image can be displayed in such that while measuring the sample, two
or more slice images in such an arrangement on a common screen that
their positions relative to the sample enables to be grasped.
Inventors: |
Ishida; Hideyuki;
(Hadano-shi, JP) ; Tanaami; Takeo; (Musashino-shi,
JP) |
Correspondence
Address: |
WESTERMAN, HATTORI, DANIELS & ADRIAN, LLP
1250 CONNECTICUT AVENUE, NW
SUITE 700
WASHINGTON
DC
20036
US
|
Assignee: |
TOKAI UNIVERSITY EDUCATIONAL
SYSTEM
Tokyo
JP
JAPAN SCIENCE AND TECHNOLOGY AGENCY
Kawaguchi
JP
YOKOGAWA ELECTRIC CORPORATION
Musashino-shi
JP
|
Family ID: |
33549133 |
Appl. No.: |
11/403832 |
Filed: |
April 14, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
10853355 |
May 26, 2004 |
|
|
|
11403832 |
Apr 14, 2006 |
|
|
|
Current U.S.
Class: |
250/201.3 ;
250/235 |
Current CPC
Class: |
G02B 21/0044 20130101;
G02B 21/008 20130101 |
Class at
Publication: |
250/201.3 ;
250/235 |
International
Class: |
G02B 27/40 20060101
G02B027/40; H01J 3/14 20060101 H01J003/14; G02B 27/64 20060101
G02B027/64 |
Foreign Application Data
Date |
Code |
Application Number |
May 27, 2003 |
JP |
2003-149125 |
Claims
1-5. (canceled)
6. A confocal microscope display device for displaying a plurality
of slice images of a sample picked up at different positions in an
optical axis direction by using an optical microscope and a
confocal scanner, comprising: a moving portion for moving an
objective lens of the optical microscope in the optical axis
direction; and a processing portion for reading the slice images of
the sample obtained from the confocal scanner, to display the slice
images on a screen, wherein while measuring the sample, the
processing portion displays two or more slice images in such an
arrangement on a common screen that their positions relative to the
sample enables to be grasped.
7. (canceled)
8. The confocal microscope display device according to claim 6,
wherein the processing portion displays the slice images
one-dimensionally or two-dimensionally in such an arrangement that
their positions relative to the sample enables to be grasped.
9. The confocal microscope display device according to claim 6,
wherein the display number of the slice images enables to be
changed while measuring the sample.
10. The confocal microscope display device according to claim 6,
wherein the slice images are presented in a perspective view so
that the processing portion displays the sample and each of the
slice images at coincident relative positions in the optical axis
direction.
11. The confocal microscope display device according to claim 6,
wherein the display angle of the perspective view of the slice
images enables to be changed while measuring the sample.
12. The confocal microscope display device according to claim 6,
wherein the processing portion displays sizes, axes, frames or
marker images over the slice images.
13. The confocal microscope display device according to claim 6,
wherein the moving portion includes a piezo-element, stage drive
portion or a magnet actuator.
14. The confocal microscope display device according to claim 6,
wherein the processing portion includes a personal computer.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] The present invention relates to a confocal microscope,
which is enabled to measure a stereoscopic shape of a sample by
combining an optical microscope and a confocal optical-scanner.
[0003] 2. Description of the Related Art
[0004] For example, a Nipkow's disc type confocal microscope
apparatus, as shown in FIG. 1A, is well known in the related art.
This confocal microscope apparatus is provided with: a microlens
array 101, a pinhole array 102 (i.e., Nipkow's disc) and an
objective lens 103 for condensing a laser light onto a sample 20;
an actuator 104 for moving the objective lens 103 in an optical
axis direction (or in a Z-direction, as shown); a camera 106 having
a condensing lens 105; and a beam splitter 107 for changing the
path of the reflected light coming from the sample through the
objective lens 103 and the pinhole array 102, in the direction to
the camera 106.
[0005] In the configuration, the Z-coordinate of the focused point
of the laser light is controlled depending on the position of the
objective lens 103 in the Z-direction, and the XY-coordinates of
the focused point of the laser light is controlled by turning the
microlens array 101 and the pinhole array 102. In other words, the
scanning point in the sample 20 to be picked up by the camera 106
can be three-dimensionally controlled depending on the Z-direction
position of the objective lens 103 and the turning angles of the
microlens array 101 and the pinhole array 102.
[0006] In the such a scanning technique of the confocal microscope
apparatus the operations to move the objective lens 103 uniformly
in a Z-coordinate increasing direction for a longer period than a
plurality of frame periods are started in synchronization with a
vertical synchronizing signal of the camera, as produced just after
the input of a trigger signal, while turning the microlens array.
101 and the pinhole array 102 in synchronization with the vertical
synchronizing signal of the camera 106. This scanning technique is
described, for example, in JP-A-2002-72102.
[0007] In the scanning technique, however, the timing for starting
the movement of the objective lens 103 is synchronized with the
vertical synchronizing signal, but the movement after the start is
performed asynchronously of the vertical synchronizing signal. As a
result, it is difficult to control the Z-direction position of the
scanning point highly precisely for the individual video frames to
be picked up by the camera 106. In the case of the repeated
capturing with the movement of the Z-direction position, more
specifically, the discrepancy of the Z-direction position is so
cumulatively enlarged that the discrepancy can be neither confirmed
nor corrected.
[0008] In the related art described above, moreover, the individual
scanning points are captured by scanning in the XY-directions while
changing the Z-coordinate at all times. According to the capturing
method by thus changing the Z-coordinate at all times, moreover,
the Z-coordinate point can be prevented from being unscanned for
all the XY-coordinates so that even a micro structure in the
Z-direction can enhance the probability of its appearance at least
in the captured images.
[0009] In the related art, the coordinates of the objective lens
103 change uniformly, too, even for the time period of the
synchronizing signal such as the vertical synchronizing signal,
when the capturing is not done in the camera 106. However, that
Z-coordinate range in the sample 20, which corresponds to the range
for the objective lens 103 to have moved for the synchronizing
signal period, is not captured in the least. According to the
related art, therefore, a micro structure in the Z-direction may
drop out.
[0010] Depending on the application of the confocal microscope
apparatus, on the other hand, the video frames having picked up the
XY-plane of the sample with the Z-coordinate being fixed may be
desirably produced individually for the different Z-coordinates.
For example, a set of video frames thus produced become as they are
the voxels having the XYZ-coordinate system so that they are suited
for the processing such as the three-dimensional analysis of the
sample 20.
[0011] According to the related art thus far described, however,
the Z-coordinate always changes, too, for the video pickup period
of the camera 106 so that the video frames having picked up the
XY-plane of the sample with the Z-coordinate being fixed cannot be
produced.
SUMMARY OF THE INVENTION
[0012] An object of the invention is to provide a confocal
microscope apparatus that improves the precision of the scanning
position control of a sample in the optical axis direction.
[0013] Another object of the invention is to provide a confocal
microscope apparatus that enhances the probability of grasping a
micro structure in an image picked up.
[0014] A further object of the invention is to provide a confocal
microscope apparatus that creates video frames captured by picking
up a plane normal to the optical axis of a sample with the
coordinate in the optical axis direction being fixed, individually
for the coordinates in the different optical axis directions.
[0015] A further object of the invention is to provide a confocal
microscope apparatus that creates video frames captured by picking
up a plane normal to the optical axis of the sample with the
coordinate in the optical axis direction being fixed, individually
for the coordinates in the different optical axis directions, and
to display a three-dimensional image at a high speed, thereby to
grasp the whole image while measuring a sample.
[0016] A further object of the invention is to provide a confocal
microscope apparatus that grasps slice images in each section and
their stereoscopic relations precisely.
BRIEF DESCRIPTION OF THE DRAWINGS
[0017] FIGS. 1A and 1B are diagrams showing a configuration and a
scanning sequence of a confocal microscope apparatus of the related
art;
[0018] FIG. 2 is a schematic diagram showing a configuration of a
confocal microscope apparatus according to an embodiment of the
invention;
[0019] FIGS. 3A to 3G are diagrams showing a scanning sequence
according to the embodiment of the invention;
[0020] FIGS. 4A and 4B are block diagrams showing examples of the
configurations of a Z-axis scan control device and an actuator
according to the embodiment of the invention;
[0021] FIGS. 5A to 5G are diagrams showing another scanning
sequence according to the embodiment of the invention;
[0022] FIG. 6 shows a display example of a three-dimensional image
according to the confocal microscope of the related art;
[0023] FIGS. 7A to 7D show map display examples of measured
images;
[0024] FIG. 8 is a configuration diagram showing another example of
the confocal microscope apparatus according to the invention;
[0025] FIG. 9 is a diagram showing a three-dimensional display
example having a plurality of slice images; and
[0026] FIGS. 10A and 10B are contrast diagrams of the map display
and a perspective display.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0027] The invention will be described in detail with reference to
the accompanying drawings. FIG. 2 is a configuration diagram
showing one embodiment of a confocal microscope apparatus according
to the invention.
[0028] As shown in FIG. 2, the confocal microscope apparatus is
provided with: a body portion 1; a stage 2 for mounting a sample
20; a laser light source 3; a confocal scanner unit 4; a high-speed
camera 5 such as a double-speed camera of 66 frames/second adopting
IEEE 1394 as the communication standards; an image intensifier 6
for adding an image intensifying function, a high-speed shutter
function and so on to that camera 5; an objective lens 7; an
actuator 8 for moving the objective lens 7 in the direction of an
optical axis; a Z-axis scan control device 9; an image processing
device 10 constructed of a computer or the like having a video
capture interface; and a display device 11. In the confocal scanner
unit 4, moreover, there are housed the microlens array, the pinhole
array, the beam splitter, as has been described hereinbefore, and
the rotation control unit for rotationally driving the microlens
array and the pinhole array.
[0029] The present confocal microscope apparatus is further
provided with an illuminating light source 12 so that it also
functions as an optical microscope apparatus with the illuminating
light source 12 and the optical system housed in the body portion
1.
[0030] The scan control in the present confocal microscope
apparatus will be described below. FIGS. 3A to 3G show a scanning
sequence of the confocal microscope apparatus. The scanning
sequence is started with a TRIGER signal outputted from the Z-axis
scan control device 9. In response to the TRIGER signal as an
external trigger signal, the camera 5 synchronizes a vertical
synchronizing signal with the TRIGER signal, as shown in FIG. 3B,
to start the capturing.
[0031] On the other hand, the vertical synchronizing signal VSYNC
of the camera 5 is outputted to the confocal scanner unit 4 and the
Z-axis scan control device 9. In synchronization with the vertical
synchronizing signal VSYNC inputted, the (not-shown) rotation
control unit of the confocal scanner unit 4 drives the microlens
array and the pinhole array so rotationally as to scan the whole
XY-area once for every image pickup periods of the individual video
frames. Here, the image pickup period portion such a period in one
video frame period as excludes at least the vertical synchronizing
signal period. Moreover, the image pickup period may exclude a
horizontal synchronizing signal period and the period, for which
the pixels before and after the horizontal synchronizing signal
period and the vertical synchronizing signal period.
[0032] In synchronization with the vertical synchronizing signal
VSYNC inputted, as shown in FIG. 3C, the Z-axis scan control device
9 outputs a predetermined number of pulses of a predetermined
period in the image pickup period of each video frame, as a
movement control signal CNT to the actuator 8. Moreover, the Z-axis
scan control device 9 counts the vertical synchronizing signals
VSYNC inputted, and stops the pulse output when a predetermined
count is reached. The Z-axis scan control device 9 executes a
stage, at which it outputs a reset signal RST to the actuator 8, as
shown in FIG. 3D. When the next vertical synchronizing signal VSYNC
is inputted, the Z-axis scan control device 9 repeats such a
sequence like before for the period of a predetermined number of
video frames as is composed of the stage, at which it outputs the
pulses of a predetermined number of predetermined periods as the
movement control signal CNT to the actuator 8, and the stage, at
which it stops the pulse output for one video frame period and at
which it outputs the reset signal RST to the actuator 8.
[0033] On the other hand, the actuator 8 integrates the pulses of
the movement control signal CNT inputted from the Z-axis scan
control device 9, to produce the drive signals which keep constant
values for the periods of the vertical synchronizing signals VSYNC
but uniformly increase for the image pickup periods of the video
frames, as shown in FIGS. 3E and 3F. The objective lens 7 is moved
in the Z-direction with those drive signals. Here, the movement of
the objective lens 7 is made proportional to the magnitudes of the
drive signals. If the movement of the objective lens 7 is not
linearly proportional to the magnitudes of the drive signals, the
drive signals are produced to have such a waveform as to move the
objective lens 7 not for the periods of the vertical synchronizing
signals VSYNC but uniformly for the image pickup periods of the
video frames. Here, the mechanism for moving the objective lens 7
can be exemplified by one using a piezo-element.
[0034] In response to the reset signal RST from the Z-axis scan
control device 9, moreover, the actuator 8 returns the drive
signals to the initial value.
[0035] By the operations thus far described, the scanning points in
the sample 20 and the individual video frames are given such
relations as are shown in FIG. 3G. For the individual video frame
periods (as indicated by T1 to T8), the images in the different
Z-axis ranges are picked up for every video frame periods. In this
embodiment, the objective lens 7 is moved in the Z-direction only
for the image pickup periods of the video frames. For any of the
XY-coordinates, therefore, there hardly occurs a Z-coordinate range
of no capturing.
[0036] Reverting to FIG. 2, the image processing device 10 repeats
the operations to fetch and store the individual video frames VIDEO
outputted from the camera 5 and to synthesize and display them in
the display device 11. Here, the image processing device 10 is fed
with timing signals indicating the timings of the TRIGER signals,
from the Z-axis scan control device 9. In accordance with these
timing signals, the image processing device 10 repeats the
operations: to recognize the correspondences between the individual
video frames and the orders of the samples 20 picked up in the
video frames, in the Z-direction of the scanning face; to
synthesize and arrange the individual video frames in accordance
with the recognized orders thereby to reconstruct the
three-dimensional images (or voxels) of the samples 20; and to
display such a three-dimensionally expressed image in the display
device 11 that the three-dimensional image is projected on a
virtual two-dimensional screen by a suitable rendering algorithm
(e.g., a volume rendering). In short, the image processing device
10 makes a real time display of the three-dimensionally expressed
image of the sample 20.
[0037] One configuration example of the Z-axis scan control device
9 will be described in the following. FIGS. 4A and 4B shows the
configuration example of the Z-axis scan control device 9. The
following description is made by assuming that the Z-axis scan
control device 9 repeats such a sequence in the scanning sequence
shown in FIGS. 3A to 3G as is composed of the stage, at which the
Z-axis scan control device 9 outputs a predetermined number M of
pulses of a predetermined period T as the movement control signal
CNT to the actuator for the image pickup period of each vide frame
in a predetermined number N of video frame periods, and the stage,
at which the Z-axis scan control device 9 stops the pulse output
for one video frame period and outputs the reset signal RST to the
actuator 8.
[0038] In FIGS. 4A and 4B, a sequence control unit 91 generates the
aforementioned TRIGER signal in response to a demand from the image
processing device 10, the user's operation or the like.
[0039] A first counter 92 is reset with the TRIGER signal to count
the vertical synchronizing signals VSYNC from 0. A first decoder 93
decodes the counted value of the first counter 92. When this
counted value reaches the predetermined value N, the first decoder
93 outputs a reset enable signal to a reset output circuit 94 and a
mask circuit 95, and outputs a counter reset signal to the first
counter 92. The reset output circuit 94 produces, when fed with the
reset enable signal, the reset signal RST of a predetermined pulse
length, and outputs the reset signal RST to the actuator 8. When
the first counter 92 is fed with a counter reset signal, on the
other hand, it is reset to 0 in synchronization with the input of
the next vertical synchronizing signal VSYNC.
[0040] A second counter 96 counts clock signals of the
predetermined period T outputted by an oscillator 97, from 0. A
second decoder 98 decodes the counted value. When this counted
value becomes M, the second decoder 98 outputs a pulse mask signal
to the mask circuit 95 and outputs a stop signal to the second
counter 96. Only for the time period while the reset enable signal
is not outputted from the first decoder 93 and while the pulse mask
signal is not outputted from the second decoder 98, the mask
circuit 95 outputs the clock signal of the predetermined period T
outputted from the oscillator 97, as the pulse of the movement
control signal CNT to the actuator 8. Here, the second counter 96
stops the counting operation, when fed with the stop signal, until
the vertical synchronizing signal VSYNC is inputted. When the
vertical synchronizing signal VSYNC is inputted, the second counter
96 resets the count value to 0, and starts the counting
operation.
[0041] The foregoing configuration of the Z-axis scan control
device 9 is just one example, and can adopt another. In a
configuration, for example, a PLL can be used to produce a pulse
signal of a 1/M period having an image pickup period synchronized
with the vertical synchronizing signal, and this pulse signal can
be outputted as the movement control signal CNT only for the image
pickup period. Alternatively, the Z-axis scan control device 9 may
also be constructed as a CPU circuit so that the foregoing
operations of the Z-axis scan control device 9 may be executed in
the software manner.
[0042] Next, the drive signal is produced in the actuator 8 by
integrating the pulses of the movement control signal CNT inputted
from the Z-axis scan control device 9, as has been described
hereinbefore. This integration may be made by the well-known analog
integration circuit. Another analog integration circuit can be
constructed, as shown in FIG. 4B, to include: a counter 81 for
counting the pulses of the movement control signal CNT; a D/A
converter 82 for D/A converting the counted value of the counter
81; and a driver circuit for amplifying the output of the D/A
converter 82. Here, the counter 81 is reset with the reset signal
RST inputted from the Z-axis scan control device 9.
[0043] Now, the confocal microscope apparatus of the embodiment
thus far described may further execute the following scanning
sequence.
[0044] As shown in FIGS. 5A to 5G, the scanning sequence is started
with the TRIGER signal, as shown in FIG. 5A, which is outputted by
the Z-axis scan control device 9. The camera 5 receives the TRIGER
signal as the external trigger signal, and synchronizes the
vertical synchronizing signal with the TRIGER signal, as shown in
FIG. 5B, to start the capturing.
[0045] The vertical synchronizing signal VSYNC of the camera 5 is
outputted to the confocal scanner unit 4 and the Z-axis scan
control device 9. In synchronization with the vertical
synchronizing signal VSYNC inputted, the rotation control unit of
the confocal scanner unit 4 drives the microlens array and the
pinhole array so rotationally as to scan the whole XY-area once for
every image pickup periods of the individual video frames.
[0046] In synchronization with the vertical synchronizing signal
VSYNC inputted, as shown in FIG. 5C, the Z-axis scan control device
9 executes the stage, at which it outputs pulses for the vertical
synchronizing signal period, as the movement control signal CNT to
the actuator 8. Moreover, the Z-axis scan control device 9 counts
the vertical synchronizing signals VSYNC inputted, and stops the
pulse output of the movement control signal CNT when a
predetermined count is reached. The Z-axis scan control device 9
executes a stage, at which it outputs the reset signal RST to the
actuator 8, as shown in FIG. 5D. When the next vertical
synchronizing signal VSYNC is inputted, the Z-axis scan control
device 9 repeats such a sequence like before for the period of a
predetermined number of video frames as is composed of the stage,
at which it outputs the pulses for the vertical synchronizing
signal period for a predetermined number of video frame periods as
the movement control signal CNT to the actuator 8, and the stage,
at which it stops the pulse output of the movement control signal
CNT for one video frame period and at which it outputs the reset
signal RST to the actuator 8.
[0047] The actuator 8 integrates the pulses of the movement control
signal CNT inputted from the Z-axis scan control device 9, to
produce the drive signals which increase for the vertical
synchronizing signal period but keep constant values for the image
pickup periods of the video frames, as shown in FIGS. 5E and 5F.
The objective lens 7 is moved in the Z-direction with those drive
signals. Here, the movement of the objective lens 7 is made
proportional to the magnitudes of the drive signals.
[0048] In response to the reset signal RST from the Z-axis scan
control device 9, moreover, the actuator 8 returns the drive
signals to the initial value.
[0049] By the operations thus far described, the scanning points in
the sample 20 and the individual video frames are given such
relations as are shown in FIG. 5G. For the individual video frame
periods (as indicated by T1 to T8), the images in the XY-plane
having a specific Z-coordinate spaced for every video frames are
picked up for every video frame periods.
[0050] According to the scanning sequence thus far described, the
objective lens 7 is moved in the Z-direction only for the image
pickup period of the video frames. Therefore, the video frames
having picked up the XY-plane of the sample 20 with the fixed
Z-coordinate can be created individually for the different
Z-coordinates.
[0051] As described hereinbefore, the confocal microscope apparatus
is enabled to improve the precision of the scanning position
control of the sample better in the optical axis direction.
Moreover, the confocal microscope apparatus is enabled to enhance
the probability of grasping a micro structure in the image picked
up. Still moreover, the confocal microscope apparatus is enabled to
create the video frames, which are picked up by picking up a plane
normal to the optical axis of the sample with the coordinate in the
optical axis direction being fixed, individually for the
coordinates in the different optical axis directions.
[0052] Another embodiment of the invention will be described in the
following. In the case a stereoscopic image of the sample is to be
attained with the confocal microscope apparatus using the confocal
scanner, a number of slice images are obtained at different
positions in the optical axis direction, as described above, and
are made stereoscopic by the CG (Computer Graphics) technique. FIG.
6 is a display example of the three-dimensional image of a
Californian purple sea urchin measured by that method. By this
display, the whole image of the sample can be grasped.
[0053] However, this case has the following problems.
[0054] (1) The CG processing takes time at least several minutes to
several hours. This image processing after the CG has to be
performed after the measurement. It is difficult to grasp the whole
image during the measurement, to decide the propriety of the sample
and to select the best portion of measurement.
[0055] (2) The shapes in the individual sections cannot be
precisely grasped with perspective views. The shapes of the
individual sections can be precisely grasped neither too much nor
too less by using the two-dimensional images (or the slice images),
as shown in FIG. 7. It is, however, difficult to grasp the
stereoscopic relations as a whole with those slice images. Here,
the slice images of FIGS. 7A, 7B, 7C and 7D correspond to the
individual slice images from up to down in the case a cell is
placed at the position of the sample. These views are binarized for
the convenience of display.
[0056] FIG. 8 is a diagram of another embodiment of the invention,
which has solved those problems. The confocal microscope apparatus
of this embodiment is enabled to display a three-dimensional image
at a high speed thereby to grasp the whole image while the sample
is being measured and to grasp the slice images in the individual
sections and their stereoscopic relations precisely in real
time.
[0057] In FIG. 8: reference numeral 100 designates an optical
microscope (as will be called merely the "microscope"); numeral 200
designates a confocal optical scanner disposed at the light
receiving portion of the microscope 100; numeral 300 designates an
image pickup camera (as will be called merely the "camera") for
picking up that image of the sample face, which is obtained through
the confocal optical scanner 200; and numeral 400 designates a
processing portion.
[0058] The processing portion 400 is provided with a display screen
410 and is enabled to read the image data outputted from the camera
300 and subject them to a predetermined processing and to display
the image on the display screen 410. A personal computer is usually
used as that processing portion 400.
[0059] Numeral 500 designates a drive portion for moving an
objective lens 110 of the microscope 100 in the optical axis
direction. For example, a piezo-element (PZT) is used as the drive
portion 500.
[0060] Numeral 600 designates a stage controller for controlling
the drive portion 500 on the basis of an instruction coming from
the processing portion 400.
[0061] Here, the components of FIG. 8 and the components of FIG. 2
correspond in the following manners. The optical microscope 100
corresponds to the body portion 1 of FIG. 2; the objective lens 110
corresponds to the objective lens 7 of FIG. 2; the confocal optical
scanner 200 corresponds to the confocal scanner unit 4 of FIG. 2;
the image pickup camera 300 corresponds to the camera 5 of FIG. 2;
the sample 20 corresponds to the sample 20 of FIG. 2; the
processing portion 400 corresponds to the image processing device
10 of FIG. 2; the screen 410 corresponds to the display device 11
of FIG. 2; the drive portion 500 corresponds to the actuator of
FIG. 2; the stage controller 600 corresponds to the Z-axis scan
control device 9 of FIG. 2.
[0062] In this configuration, the operations to obtain the slice
images of the sample 20 placed on the microscope 100 are identical
to those of the confocal microscope apparatus of the related art,
and their description is omitted.
[0063] While the objective lens 110 is moved in the optical axis
direction by activating the drive portion 500, the confocal slice
images are picked up at the individual optical axis heights by the
camera 300. The processing portion 400 transforms the images (in
the top plan view) obtained from the camera 300 into the
perspective images (or the corresponding images) picked up
obliquely downward, and display them on the screen 410.
[0064] These transformations into the perspective views may be made
merely by drawing pixels of coordinates Xi and Yi at the plane
coordinates Xj and Yj of a predetermined perspective view, so that
the transformations can be processed at a high speed.
[0065] For images of an inclination of 30 degrees, the coordinates
Xj and Yj are determined, for example, on the basis of the
following Formulas: Xi=Xj cos .theta.-Yj sin .theta.; Yi=Xj sin
.theta.-Yj cos .theta.,
[0066] wherein .theta.=30.degree..
[0067] The coordinates Xj and Yj can be determined merely by the
product/sum operations, if the processing portion 400 has the cos
30.degree. as the table of constants. The product/sum operations
can be processed at high speeds.
[0068] In the case a plurality of slice images are to be displayed,
they are drawn as they are at a spacing in the optical axis
direction while being held at their relative positions in the
optical axis direction, as shown in FIG. 9. FIG. 9 is an example of
the image display of the case, in which the measurement and the
display are actually performed in real time.
[0069] FIG. 9 shows the motions of calcium ions in the muscle of
heart, in which a white bright spot moves from the left depth of
the screen to this side. With these four images, it can be
intuitively grasped at a glance that the calcium ions spread
earlier in the cell of the uppermost slice image than the lowermost
slice image. This makes it possible not only to analyze the data
after acquired but also either to decide the propriety of what
sample is to be actually measured, or to select the best portion of
measurement.
[0070] Thus, the confocal microscope apparatus of this embodiment
can grasp the precise slice images of the sample at the individual
optical axis heights and the stereoscopic relations of the samples
as a whole.
[0071] The invention may be exemplified by the
changes/modifications, as will be enumerated in the following.
[0072] (1) In the case a plurality of slice images are to be
obtained, the XY-plane of the sample may be captured by the
aforementioned scanning sequence with the Z-coordinate being
fixed.
[0073] (2) The number of display sheets should not be limited to
four but can be any from two to several tens.
[0074] (3) The display angle can be 0 to 360.degree. individually
in the longitudinal and latitudinal directions.
[0075] (4) For the image display, all the images need not be
displayed, but some may be thinned out. For example, the confocal
optical scanner can raise the speed up to 1,000 sheets/second, but
the display cannot be recognized by the human eyes even if it is
made at a speed exceeding a human-recognizable video rate (about 30
sheets/second) In this case, the display of one sheet per
1,000/30=33 (sheets) is sufficient.
[0076] (5) Alternatively, the image display need not display all
the slice images being measured but may display only a
representative image, as shown in FIG. 9. This display method is
more advantageous in the high speed and the recognition than the
aforementioned display method (3).
[0077] FIGS. 10A and 10B present contrast diagrams of the cases, in
which the slice images of Ca ions in the cells of the muscle of
heart are displayed in different formats. FIG. 10A presents the map
displays shown in FIGS. 7A to 7D, and FIG. 10B presents the display
example of the perspective view formats according to the invention.
Here, the displays (1) to (4) of FIG. 10A correspond to the
displays (1) to (4) of FIG. 10B.
[0078] As shown in FIG. 10B, the arrangement is devised to display
the slice images on the common screen so that the positions of the
slice images relative to the sample can be grasped. Then, it is
found that FIG. 10B presents a stereoscopically more recognizable
image display than FIG. 10A.
[0079] (6) The display image should not be limited to a
monochromatic display but may be a multicolor display.
[0080] (7) The measurement of sizes and the grasp of shapes are
facilitated if known markers such as graduations or circles or
known scales are displayed together with the slice images.
[0081] (8) Even the map display format shown in FIGS. 7A to 7D can
be utilized for deciding the propriety of the sample to some extent
although its stereoscopic grasp is difficult, if the display can be
made in real time.
[0082] (9) The drive of the objective lens 110 should not be
limited to that of the piezo-element but may be exemplified by a
stage drive or that of a magnetic actuator.
[0083] (10) The sample 20 should not be limited to a living
organism with a fluorescent light but may be a semiconductor
surface or a mechanical part with a reflecting mirror.
[0084] (11) A more proper display can be obtained if the angle or
number of displays can be changed during the
measurement/display.
[0085] (12) The image display may be updated for each slice image
at any time when the slice image is measured, or the slice images
displayed in the display screen may be updated all at once when
their measurement was ended.
[0086] According to the confocal microscope apparatus of the
embodiment shown in the configuration diagram of FIG. 8, as
described hereinbefore, the following effects can be obtained.
[0087] (1) The three-dimensional display at a high speed can be
easily realized to grasp the whole image easily while the sample is
being measured.
[0088] (2) It is possible to grasp the slice images of the
individual sections and their stereoscopic relations precisely.
* * * * *