U.S. patent application number 15/679478 was filed with the patent office on 2018-12-06 for optical sectioning apparatus using advanced optical interference microscopy.
The applicant listed for this patent is AcuSolutions Inc.. Invention is credited to Kuang-Yu HSU, Chien-Chung TSAI.
Application Number | 20180347960 15/679478 |
Document ID | / |
Family ID | 61163581 |
Filed Date | 2018-12-06 |
United States Patent
Application |
20180347960 |
Kind Code |
A1 |
TSAI; Chien-Chung ; et
al. |
December 6, 2018 |
OPTICAL SECTIONING APPARATUS USING ADVANCED OPTICAL INTERFERENCE
MICROSCOPY
Abstract
An optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy, including: a beam splitter
capable of splitting an incident light beam into a reflected light
beam and a transmitted light beam; a wide band light source for
providing the incident light beam; a reference arm unit for making
the transmitted light beam travel a round trip along an adjustable
optical path; a short wavelength light source; a first dichroic
splitter, with a first side facing the short wavelength light
source and a third side facing the beam splitter, being capable of
providing a light-blocking effect on wavelengths shorter than a
preset wavelength; an objective lens, with a collimated side facing
a second side of the first dichroic splitter; a sample carrier unit
facing a focal side of the objective lens; and a projection lens
and a sensor units for receiving an output light beam.
Inventors: |
TSAI; Chien-Chung; (Taipei
City, TW) ; HSU; Kuang-Yu; (Taipei City, TW) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AcuSolutions Inc. |
Apia |
|
WS |
|
|
Family ID: |
61163581 |
Appl. No.: |
15/679478 |
Filed: |
August 17, 2017 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01B 9/02065 20130101;
G01B 9/02041 20130101; G01B 9/02057 20130101; G02B 21/16 20130101;
G01B 9/02091 20130101; G02B 21/367 20130101; G01B 9/0203 20130101;
G02B 21/0056 20130101; H04N 7/18 20130101; G01B 2290/70 20130101;
G01B 2290/60 20130101; G02B 21/14 20130101; A61B 5/0066 20130101;
G02B 21/0076 20130101 |
International
Class: |
G01B 9/02 20060101
G01B009/02; H04N 7/18 20060101 H04N007/18; A61B 5/00 20060101
A61B005/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 5, 2017 |
TW |
106118559 |
Claims
1. An optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy, including: a beam splitter,
having a first side, a second side, a third side, and a fourth
side, and being capable of splitting an incident light beam
received from the first side into a reflected light beam traveling
out the second side and a transmitted light beam traveling out the
third side; a wide band light source apparatus for generating a
wide band light beam to be incident on the first side of the beam
splitter; a reference arm unit, used to make the transmitted light
beam travel a round trip along an adjustable optical path and then
return to the beam splitter; a short wavelength light source
apparatus for generating a short wavelength light beam; a first
dichroic splitter, having a first side, a second side, and a third
side, with the first side facing the short wavelength light source
apparatus, the third side facing the second side of the beam
splitter, and being capable of providing a light-blocking effect on
a band of wavelengths shorter than a preset wavelength, and the
short wavelength light beam having a wavelength shorter than the
preset wavelength; a first objective lens, having a collimated side
and a focal side, with the collimated side facing the second side
of the first dichroic splitter; a sample carrier unit facing the
focal side of the first objective lens and being used for carrying
a sample stained with a fluorescent dye; a projection lens, having
a light entrance side and a light exit side, with the light
entrance side facing the fourth side of the beam splitter; and a
sensor unit facing the light exit side of the projection lens.
2. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 1,
wherein the reference arm unit includes: an optical path delay
unit, having a first side and a second side, with the first side
facing the third side of the beam splitter; a second objective
lens, having a collimated side and a focal side, with the
collimated side facing the second side of the optical path delay
unit; and a reflective mirror, facing the focal side of the second
objective lens and used for reflecting the transmitted light beam,
wherein the optical path delay unit is used for adjusting the
adjustable optical path so that the adjustable optical path and a
sample arm optical path have an optical path difference less than a
coherent length, with the sample arm optical path being formed by
the sample carrier unit, the first objective lens, and the first
dichroic splitter.
3. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 1,
wherein both the wide band light source apparatus and the short
wavelength light source apparatus are implemented with a light
source, or a combination of a light source and a grating, or a
combination of a light source, a grating, and a tilt-adjustable
reflective mirror, or a plurality of parallel LED stripes.
4. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 1,
wherein the sample carrier unit further includes a white light
source to provide a proper intensity of white light to pass through
the first objective lens, the white light source including a
white-light LED, a white-light halogen lamp, or a tungsten
lamp.
5. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 1,
wherein the sensor unit includes a second dichroic splitter, a
two-dimensional color sensing device, a long wavelength pass
filter, and a two-dimensional monochrome sensing device, in which
the second dichroic splitter has a first side, a second side, and a
third side, with the first side facing the projection lens, and the
second dichroic splitter is used to reflect a fluorescent light
beam and a white light beam to and through the third side to form
an image on the two-dimensional color sensing device, and to
transmit a wide band light beam through the second side to form an
image on the two-dimensional monochrome sensing device; or the
sensor unit includes a flip-type reflective mirror, a
two-dimensional color sensing device, a long wavelength pass
filter, and a two-dimensional monochrome sensing device, in which
the flip-type reflective mirror has a flip-on state to allow a
white light beam to form an image on the two-dimensional color
sensing device, and has a flip-off state to allow a wide band light
beam and a fluorescent light beam to form an image on the
two-dimensional monochrome sensing device, and the long wavelength
pass filter can be disposed between the projection lens and the
flip-type reflective mirror, or be disposed between the projection
lens and the second dichroic splitter, or be disposed between the
flip-type reflective mirror and the two-dimensional monochrome
sensing device, or be disposed between the second dichroic splitter
and the two-dimensional monochrome sensing device.
6. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 5,
wherein the wide band light beam has a wavelength range of 470
nm-800 nm, the short wavelength light beam has a wavelength range
of 365 nm-460 nm, the beam splitter has an operating wavelength
range of 400 nm-800 nm, and both the first dichroic splitter and
the long wavelength pass filter have a cut-off wavelength range of
400 nm-470 nm.
7. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 5,
wherein the wide band light beam has a wavelength range of 650
nm-1000 nm, the short wavelength light beam has a wavelength range
of 365 nm-630 nm, the beam splitter has an operating wavelength
range of 400 nm-1000 nm, and all the first dichroic splitter, the
second dichroic splitter, and the long wavelength pass filter have
a cut-off wavelength range of 400 nm-650 nm.
8. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 2,
wherein a first polarizer is disposed between the wide band light
source apparatus and the beam splitter, a second polarizer is
disposed in front of the two-dimensional monochrome sensing device,
a first quarter-wave plate is disposed between the first objective
lens and the first dichroic splitter, and a second quarter-wave
plate is disposed between the optical path delay unit and the
second objective lens, wherein the first polarizer has a first
polarization direction, the second polarizer has a second
polarization direction orthogonal to the first polarization
direction, the first quarter-wave plate has a first optical axis
direction, the second quarter-wave plate has a second optical axis
direction, and both the first optical axis direction and the second
optical axis direction are disposed within a range formed by the
first polarization direction and the second polarization direction
to enhance an interference effect and promote an imaging
quality.
9. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 5,
wherein a first polarizer is disposed between the wide band light
source apparatus and the beam splitter, a second polarizer is
disposed in front of the two-dimensional monochrome sensing device,
a first quarter-wave plate is disposed between the first objective
lens and the first dichroic splitter, and a second quarter-wave
plate is disposed between the optical path delay unit and the
second objective lens, wherein the first polarizer has a first
polarization direction, the second polarizer has a second
polarization direction orthogonal to the first polarization
direction, the first quarter-wave plate has a first optical axis
direction, the second quarter-wave plate has a second optical axis
direction, and both the first optical axis direction and the second
optical axis direction are disposed within a range formed by the
first polarization direction and the second polarization direction
to enhance an interference effect and promote an imaging
quality.
10. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 2,
further including an information processing apparatus to perform an
image processing process.
11. The optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy as disclosed in claim 10,
wherein the reference arm unit further includes an axial motion
platform, and the sample carrier unit further includes a 3-D motion
platform, so that by using the axial motion platform to move the
second objective lens and the reflective mirror, by adjusting the
optical path delay unit, and by using the 3-D motion platform to
move the sample stained with the fluorescent dye, the information
processing apparatus can derive a 3-D image of the sample
accordingly.
Description
BACKGROUND OF THE INVENTION
Field of the Invention
[0001] The present invention relates to an optical sectioning
apparatus, especially to an optical sectioning apparatus using
optical interference microscopy and fluorescence microscopy.
Description of the Related Art
[0002] In conventional tumor surgery, it takes a lot of time for a
pathologist to examine a frozen section to determine if a tumor is
cleanly removed. As a result, when the time allowed for the surgery
is shorter than the time needed for the pathologist to complete the
assessment of intraoperative pathological margin, the tumor cannot
be guaranteed to be cleanly removed.
[0003] During the process of preparing a frozen section of a
sample, if the sample contains much water, a formed crystal ice
structure will cause a damage to the tissue structure of the
sample; and if the sample contains much fat, the fat will still be
not frozen at temperatures around -20.degree. C. where the other
tissues are already frozen, and is therefore easy to slip off from
the frozen section to cause an artifact to the frozen section.
[0004] OCT (optical coherence tomography) is a newly developed
optical imaging technology, which utilizes an operational principle
similar to that of the supersonic imaging technology but has a
higher resolution than the supersonic imaging technology. OCT
mainly utilizes the phenomena that different tissues of a sample
have different responses with regard to light reflection, light
absorption, and light scattering and makes use of an optical
interference methodology to form an image of the sample for
diagnosis. Since OCT can be performed by directly scanning a sample
at room temperature without the need of including a freezing
procedure and a slicing procedure, morphological artifacts induced
by slicing a frozen sample containing much water or fat can be
prevented, the completeness of the sample can be maintained to
ensure the accuracy of a pathological assessment, the surgery time
can be shortened, and surgery effect can be improved. However,
since traditional optical microscopy usually has a wide depth of
focus, a sample needs to undergo a real sectioning procedure to
provide sections of a depth of 4-5 .mu.m, so as to avoid
overlapping of tissue images of different depths, and thereby to
derive a clear image.
[0005] To solve the foregoing problems, the U.S. Pat. No. 9,185,357
B2 discloses an optical tissue sectioning apparatus using
full-field optical coherence tomography, which includes a
full-field imaging interferometer and an optical sectioning imaging
system. This optical tissue sectioning apparatus uses the
interferometer to solve the problem resulting from the wide depth
of focus by directly deriving the image from the sample, which has
a resolution as precise as less than 1 .mu.m both laterally and
vertically, getting rid of the need of a fixation process, which
includes a freezing procedure, a paraffin-embadded procedure, and a
real sectioning procedure.
[0006] Besides, as a conventional H&E stain section uses
hematoxylin to color the nuclei bluish violet and uses eosin to
color cytoplasm pink, the nuclei will also be represented in bluish
violet in a derived image, and the details (such as the nucleolus
or heterochromatin) of each of the nuclei will be undistinguishable
in the derived image. Therefore, U.S. Pat. No. 9,185,357 B2 adopts
a fluorescent staining approach to exhibit an image of the details
of nuclei to fulfill the need of pathology inspection.
[0007] However, there is still room for improvement in the
performance of U.S. Pat. No. 9,185,357 B2, for example: (1) a
short-wavelength light beam (such as UV light beam) will be
attenuated by a beam-splitting film of the beam splitter by 60-90%;
(2) the antireflection film of the beam splitter generally has a
low transmission rate for a UV band of wavelengths shorter than 400
nm. Due to these two disadvantages, a fluorescent signal out of a
sample will very weak. To ensure the strength of a fluorescent
signal, shorten exposure time, and speed up an image taking
process, a novel optical sectioning apparatus is needed.
SUMMARY OF THE INVENTION
[0008] One objective of the present invention is to disclose an
optical sectioning apparatus, in which a first dichroic splitter is
disposed between a beam splitter and a first objective lens to
block a short-wavelength light beam and thereby increase a relative
strength of a fluorescent signal, shorten an exposure time, and
speed up taking images.
[0009] Another objective of the present invention is to disclose an
optical sectioning apparatus, in which a first dichroic splitter
will filter out a short-wavelength light beam so that when a
fluorescent light beam emitted from a sample passes through the
first dichroic splitter, the fluorescent light beam will have a
good relative strength to shorten an exposure time and speed up
taking images.
[0010] Still another objective of the present invention is to
disclose an optical sectioning apparatus, in which a sensor unit
has a long-wavelength-pass filter to further filter out a
short-wavelength light beam to increase the relative strength of a
fluorescent signal, and thereby shorten an exposure time and speed
up taking images.
[0011] To attain the foregoing objectives, an optical sectioning
apparatus using optical interference microscopy and fluorescence
microscopy is proposed, including:
[0012] a beam splitter, having a first side, a second side, a third
side, and a fourth side, and being capable of splitting an incident
light beam entering received from the first side into a reflected
light beam traveling out the second side and a transmitted light
beam traveling out the third side;
[0013] a wide band light source apparatus for generating a wide
band light beam to be incident on the first side of the beam
splitter;
[0014] a reference arm unit, used to make the transmitted light
beam travel a round trip along an adjustable optical path and then
return to the beam splitter;
[0015] a short wavelength light source apparatus for generating a
short wavelength light beam;
[0016] a first dichroic splitter, having a first side, a second
side, and a third side, with the first side facing the short
wavelength light source apparatus, the third side facing the second
side of the beam splitter, and being capable of providing a
light-blocking effect on a band of wavelengths shorter than a
preset wavelength, and the short wavelength light beam having a
wavelength shorter than the preset wavelength;
[0017] a first objective lens, having a collimated side and a focal
side, with the collimated side facing the second side of the first
dichroic splitter;
[0018] a sample carrier unit facing the focal side of the first
objective lens and being used for carrying a sample stained with a
fluorescent dye;
[0019] a projection lens, having a light entrance side and a light
exit side, with the light entrance side facing the fourth side of
the beam splitter; and a sensor unit facing the light exit side of
the projection lens.
[0020] In one embodiment, the reference arm unit includes: an
optical path delay unit, having a first side and a second side,
with the first side facing the third side of the beam splitter; a
second objective lens, having a collimated side and a focal side,
with the collimated side facing the second side of the optical path
delay unit; and a reflective mirror, facing the focal side of the
second objective lens and used for reflecting the transmitted light
beam, wherein the optical path delay unit is used for adjusting the
adjustable optical path so that the adjustable optical path and a
sample arm optical path have an optical path difference less than a
coherent length, with the sample arm optical path being formed by
the sample carrier unit, the first objective lens, and the first
dichroic splitter.
[0021] In one embodiment, both the wide band light source apparatus
and the short wavelength light source apparatus are implemented
with a light source, or a combination of a light source and a
grating, or a combination of a light source, a grating, and an
tilt-adjustable reflective mirror, or a plurality of parallel LED
stripes.
[0022] In one embodiment, the sample carrier unit further includes
a white light source to provide a proper intensity of white light
to pass through the first objective lens, the white light source
including a white-light LED, a laser-diode-pumped crystal rod, a
laser-diode-pumped crystal fiber, a white-light halogen lamp, or a
tungsten lamp.
[0023] In one embodiment, the sensor unit includes a second
dichroic splitter, a two-dimensional color sensing device, a long
wavelength pass filter, and a two-dimensional monochrome sensing
device, in which the second dichroic splitter has a first side, a
second side, and a third side, with the first side facing the
projection lens, and the second dichroic splitter is used to
reflect a fluorescent light beam and a white light beam to and
through the third side to form an image on the two-dimensional
color sensing device, and to transmit a wide band light beam
through the second side to form an image on the two-dimensional
monochrome sensing device.
[0024] In one embodiment, the sensor unit includes a flip-type
reflective mirror, a two-dimensional color sensing device, a long
wavelength pass filter, and a two-dimensional monochrome sensing
device, in which the flip-type reflective mirror has a flip-on
state to allow a white light beam to form an image on the
two-dimensional color sensing device, and has a flip-off state to
allow a wide band light beam and a fluorescent light beam to form
an image on the two-dimensional monochrome sensing device, and the
long wavelength pass filter can be disposed between the projection
lens and the flip-type reflective mirror, or be disposed between
the projection lens and the second dichroic splitter, or be
disposed between the flip-type reflective mirror and the
two-dimensional monochrome sensing device, or be disposed between
the second dichroic splitter and the two-dimensional monochrome
sensing device.
[0025] In one embodiment, the wide band light beam has a wavelength
range of 470 nm-800 nm, the short wavelength light beam has a
wavelength range of 365 nm-460 nm, the beam splitter has an
operating wavelength range of 400 nm-800 nm, and both the first
dichroic splitter and the long wavelength pass filter have a
cut-off wavelength range of 400 nm-470 nm.
[0026] In one embodiment, the wide band light beam has a wavelength
range of 650 nm-1000 nm, the short wavelength light beam has a
wavelength range of 365 nm-630 nm, the beam splitter has an
operating wavelength range of 400 nm-1000 nm, and all the first
dichroic splitter, the second dichroic splitter, and the long
wavelength pass filter have a cut-off wavelength range of 400
nm-650 nm.
[0027] In one embodiment, a first polarizer is disposed between the
wide band light source apparatus and the beam splitter, a second
polarizer is disposed in front of the two-dimensional monochrome
sensing device, a first quarter-wave plate is disposed between the
first objective lens and the first dichroic splitter, and a second
quarter-wave plate is disposed between the optical path delay unit
and the second objective lens, wherein the first polarizer has a
first polarization direction, the second polarizer has a second
polarization direction orthogonal to the first polarization
direction, the first quarter-wave plate has a first optical axis
direction, the second quarter-wave plate has a second optical axis
direction, and both the first optical axis direction and the second
optical axis direction are disposed within a range formed by the
first polarization direction and the second polarization direction
to enhance an interference effect and promote an imaging
quality.
[0028] In one embodiment, an information processing apparatus is
used to perform an image processing process.
[0029] In one embodiment, the reference arm unit further includes
an axial motion platform, and the sample carrier unit further
includes a 3-D motion platform, so that by using the axial motion
platform to move the second objective lens and the reflective
mirror of the reference arm unit, using the 3-D motion platform to
move the sample stained with a fluorescent dye, and adjusting the
optical path delay unit, the information processing apparatus can
derive a 3-D image of the sample accordingly.
[0030] To make it easier for our examiner to understand the
objective of the invention, its structure, innovative features, and
performance, we use preferred embodiments together with the
accompanying drawings for the detailed description of the
invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] FIG. 1 illustrates a block diagram of an optical sectioning
apparatus using optical interference microscopy and fluorescence
microscopy according to one embodiment of the present
invention.
[0032] FIG. 2 illustrates a beam splitting operation and a focusing
operation of the optical sectioning apparatus of FIG. 1.
[0033] FIG. 3 illustrates a block diagram of a reference arm unit
of FIG. 1 according to one embodiment of the present invention.
[0034] FIG. 4 illustrates a beam combining operation and an image
projecting operation of the optical sectioning apparatus of FIG.
1.
[0035] FIG. 5a illustrates a block diagram of a sensor unit of FIG.
1 according to one embodiment of the present invention.
[0036] FIG. 5b illustrates a block diagram of a sensor unit of FIG.
1 according to another embodiment of the present invention.
[0037] FIG. 5c illustrates a block diagram of a sensor unit of FIG.
1 according to another embodiment of the present invention.
[0038] FIG. 5d illustrates a block diagram of a sensor unit of FIG.
1 according to still another embodiment of the present
invention.
[0039] FIG. 6 illustrates a block diagram of an optical sectioning
apparatus using optical interference microscopy and fluorescence
microscopy according to another embodiment of the present
invention.
[0040] FIG. 7 illustrates a block diagram of an optical sectioning
apparatus using optical interference microscopy and fluorescence
microscopy according to still another embodiment of the present
invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0041] Please refer to FIG. 1-2, in which, FIG. 1 illustrates a
block diagram of an optical sectioning apparatus using optical
interference microscopy and fluorescence microscopy according to
one embodiment of the present invention, and FIG. 2 illustrates a
beam splitting operation and a focusing operation of the optical
sectioning apparatus in FIG. 1.
[0042] As illustrated in the figures, the optical sectioning
apparatus using optical interference microscopy and fluorescence
microscopy includes: a wide band light source apparatus 100, a
short wavelength light source apparatus 200, a beam splitter 300, a
first dichroic splitter 400, a first objective lens 500, a sample
carrier unit 600, a projection lens 700, a reference arm unit 800,
and a sensor unit 900.
[0043] The wide band light source apparatus 100 is used to generate
a wide band light beam 10 (indicated by a hollow arrow); the short
wavelength light source apparatus 200 is used to generate a short
wavelength light beam 20 (indicated by a solid arrow); the sample
carrier unit 600 is used to carry a sample 610 stained with a
fluorescent dye, which will emit a fluorescent light beam 30
(indicated by a dashed line arrow) when the short wavelength light
beam 20 is incident on it; and the sample carrier unit 600 further
includes a white light source 620 to provide a white light beam 40
(indicated by a hollow arrowhead) having a proper intensity to pass
through the first objective lens 500.
[0044] The beam splitter 300 has a first side S301, a second side
S302, a third side S303, and a fourth side S304. The wide band
light source apparatus 100 is used for generating the wide band
light beam 10 to be incident on the first side S301 of the beam
splitter 300 and divided by the beam splitter 300 into a reflected
wide band light beam traveling out the second side S302, and a
transmitted wide band light beam traveling out the third side
S303.
[0045] The short wavelength light source apparatus 200 is used to
generate the short wavelength light beam 20. The first dichroic
splitter 400 has a first side S401, a second side S402, and a third
side S403, with the first side S401 facing the short wavelength
light source apparatus 200 and reflecting the short wavelength
light beam 20, and with the third side S403 facing the second side
S302 of the beam splitter 300 to transmit a branch of the wide band
light beam 10 from the beam splitter 300 through the second side
S402.
[0046] The first objective lens 500 has a collimated side S501 and
a focal side S502, wherein, the collimated side S501 facing the
second side S402 of the first dichroic splitter 400. The sample
carrier unit 600 faces the focusing side S502 of the first
objective lens 500 and is used to carry the sample 610 stained with
a fluorescent dye. Besides, the sample carrier unit 600, the first
objective lens 500, and the first dichroic splitter 400 together
form a sample arm optical path.
[0047] The projection lens 700 has a light entrance side S701 and a
light exit side S702, with the light entrance side S701 facing the
fourth side S304 of the beam splitter 300. The sensor unit 900
faces the light exit side S702 of the projection lens 700.
[0048] Besides, the white light source 620 includes a white-light
LED, a white-light halogen lamp, or a tungsten lamp, etc.; the
first objective lens 500 has an operating wavelength range of 350
nm-1000 nm; and both the wide band light source apparatus 100 and
the short wavelength light source apparatus 200 can be implemented
with a light source, or a combination of a light source and a
grating, or a combination of a light source, a grating, and a
tilt-adjustable reflective mirror, or a plurality of parallel LED
stripes (all not shown in the figure). As the light source, the
grating, the tilt-adjustable reflective mirror, and the parallel
LED stripes are already known in prior art, they will not be
further addressed.
[0049] Please refer to FIG. 3, which illustrates a block diagram of
a reference arm unit of FIG. 1 according to one embodiment of the
present invention.
[0050] As illustrated in FIG. 3, the reference arm unit 800
includes: an optical path delay unit 810, a second objective lens
820, and a reflective mirror 830.
[0051] The optical path delay unit 810, the second objective lens
820, and the reflective mirror 830 together form an adjustable
optical path, and the reference arm unit 800 is used to make a
branch of the wide band light beam 10 travel a round trip along the
adjustable optical path and then return to the beam splitter 300
(not shown in this figure).
[0052] The optical path delay unit 810 has a first side S811 and a
second side S812, with the first side S811 facing the third side
S303 of the beam splitter 300 (not shown in this figure); the
second objective lens 820 has a collimated side S821 and a focal
side S822, with the collimated side S821 facing the second side
S812 of the optical path delay unit 810; and the reflective mirror
830 faces the focusing side S822 of the second objective lens 820
for reflecting the branch of the wide band light beam 10. In
addition, the second objective lens 820 can have an operating
wavelength range of 350-1000 nm.
[0053] The optical path delay unit 810 can use a reflective mirror
to help make the adjustable optical path and the sample arm optical
path have an optical path difference less than a coherent length by
changing a displacement of the reflective mirror. One or more
continuous interference carrier waves can be continuously recorded
by a two-dimensional monochrome sensing device 940, and a plurality
of recorded intensity values then undergo a root-mean-square
calculation and an averaging calculation to produce an en-face
interference intensity image, wherein the continuous interference
carrier wave can also be generated by a 3-D motion platform 630, of
which the principle is known in prior art, so it will not further
addressed.
[0054] Please refer to FIG. 4, which illustrates a beam combining
operation and an image projecting operation of the optical
sectioning apparatus of FIG. 1.
[0055] As illustrated in FIG. 4, the sample carrier unit 600 is
capable of reflecting the short wavelength light beam 20 and a
branch of the wide band light beam 10, and the sample 610 stained
with a fluorescent dye (not shown in this figure) will emit the
fluorescent light beam 30 when hit by the short wavelength light
beam 20. In addition, the white light source 620 (not shown in this
figure) of the sample carrier unit 600 will emit the white light
beam 40.
[0056] The first dichroic splitter 400 is used to block a band of
wavelengths shorter than a preset wavelength, and the short
wavelength light beam 20 has a wavelength shorter than the preset
wavelength, so the short wavelength light beam 20 will be blocked
by the first dichroic splitter 400, while the branch of the wide
band light beam 10, the fluorescent light beam 30, and the white
light beam 40 will pass through the first dichroic splitter
400.
[0057] The third side S303 of the beam splitter 300 faces the
reference arm unit 800 and is used to reflect the branch of the
wide band light beam 10 from the reference arm unit 800 to the
fourth side S304, and the second side S302 faces the first dichroic
splitter 400 and is used to transmit another branch of the wide
band light beam 10, the fluorescent light beam 30, and the white
light beam 40 to the fourth side S304.
[0058] Since the adjustable optical path and the sample optical
path have an optical path difference less than a coherent length,
the wideband light beam traveling a round trip along the adjustable
optical path will have an optical interference with the wideband
light beam traveling a round trip along the sample optical path
when they meet each other. Since the interference principle is
known in prior art, it will not be addressed further.
[0059] The projection lens 700 has a light entrance side S701 and a
light exit side S702, with the light entrance side S701 facing the
fourth side S304 of the beam splitter 300, and the light exit side
S702 being used to transmit a coherent-interference resultant light
beam of the wide band light beam 10, the fluorescent light beam 30,
and the white light beam 40 to the sensor unit 900.
[0060] Please refer to FIG. 5a-5b, in which, FIG. 5a illustrates a
block diagram of a sensor unit of FIG. 1 according to one
embodiment of the present invention and FIG. 5b illustrates a block
diagram of a sensor unit of FIG. 1 according to another embodiment
of the present invention.
[0061] The sensor unit 900 includes a flip-type reflective mirror
915, a two-dimensional color sensing device 920, a long wavelength
pass filter 930 and a two-dimensional monochrome sensing device
940.
[0062] The long wavelength pass filter 930 is used to further
filter out the short wavelength light beam 20 (not shown in the
figure), and the flip-type reflective mirror 915 has a flip mount
(not shown in the figure) for presenting a flip-on state or a
flip-off state. Since the flip-type reflective mirror is known in
prior art, it will not be addressed further.
[0063] As illustrated in FIG. 5a, the long wavelength pass filter
930 is disposed between the projection lens 700 and the flip-type
reflective mirror 915. When the flip-type reflective mirror 915 is
in the flip-on state, the white light beam 40 will form an image on
the two-dimensional color sensing device 920; when the flip-type
reflective mirror 915 is in the flip-off state, the fluorescent
light beam 30 and a coherent-interference resultant light beam of
the wide band light beam 10 will form an image on the
two-dimensional monochrome sensing device 940, respectively.
[0064] As illustrated in FIG. 5b, the long wavelength pass filter
930 is disposed between the flip-type reflective mirror 915 and the
two-dimensional monochrome sensing device 940. When the flip-type
reflective mirror 915 is in the flip-on state, the white light beam
40 will form an image on the two-dimensional color sensing device
920; when the flip-type reflective mirror 915 is in the flip-off
state, the fluorescent light beam 30 and a coherent-interference
resultant light beam of the wide band light beam 10 will form an
image on the two-dimensional monochrome sensing device 940
respectively.
[0065] Please refer to FIG. 5c-5d, in which FIG. 5c illustrates a
block diagram of a sensor unit of FIG. 1 according to another
embodiment of the present invention, and FIG. 5d illustrates a
block diagram of a sensor unit of FIG. 1 according to still another
embodiment of the present invention.
[0066] The sensor unit 900 includes a second dichroic splitter 910,
a two-dimensional color sensing device 920, a long wavelength pass
filter 930 and a two-dimensional monochrome sensing device 940, in
which the long wavelength pass filter 930 is used to further filter
out the short wavelength light beam 20 (not shown in the
figure).
[0067] As illustrated in FIG. 5c, the long wavelength pass filter
930 is disposed between the projection lens 700 and the second
dichroic splitter 910. The second dichroic splitter 910 reflects
the fluorescent light beam 30 and the white light beam 40 to the
third side S903 to form an image on the two-dimensional color
sensing device 920, and transmits a coherent-interference resultant
light beam of the wide band light beam 10 through the second side
S902 to form an image on the two-dimensional monochrome sensing
device 940.
[0068] As illustrated in FIG. 5d, the long wavelength pass filter
930 is disposed between the second dichroic splitter 910 and the
two-dimensional monochrome sensing device 940. The second dichroic
splitter 910 reflects the fluorescent light beam 30 and the white
light beam 40 to the third side S903 to form an image on the
two-dimensional color sensing device 920, and transmits a
coherent-interference resultant light beam of the wide band light
beam 10 through the second side S902 to form an image on the
two-dimensional monochrome sensing device 940.
[0069] When the wide band light beam 10 has a wavelength range of
470 nm-800 nm, the flip-type reflective mirror 915 is required, the
short wavelength light beam 20 (not shown in this figure) has a
wavelength range of 365 nm-460 nm, the beam splitter 300 (not shown
in this figure) has an operating wavelength range of 400 nm-800 nm,
and the first dichroic splitter 400 has a cut-off wavelength range
of 400 nm-470 nm.
[0070] When the wide band light beam 10 has a wavelength range of
650 nm-1000 nm, the flip-type reflective mirror 915 or the second
dichroic splitter 910 can be adopted, the short wavelength light
beam 20 has a wavelength range of 365 nm-630 nm, the beam splitter
300 has an operating wavelength range of 400 nm-1000 nm, and all
the first dichroic splitter 400, the second dichroic splitter 910,
and the long wavelength pass filter 930 have a cut-off wavelength
range of 400 nm-650 nm.
[0071] Please refer to FIG. 6, which illustrates a block diagram of
an optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy according to another
embodiment of the present invention.
[0072] As illustrated in FIG. 6, a first polarizer 950 is disposed
between the wide band light source apparatus 100 and the beam
splitter 300, a second polarizer 960 is disposed in front of the
two-dimensional monochrome sensing device 940, a first quarter-wave
plate 970 is disposed between the first objective lens 500 and the
first dichroic splitter 400, and a second quarter-wave plate 980 is
disposed between the optical path delay unit 810 and the second
objective lens 820.
[0073] The first polarizer 950 has a first polarization direction,
the second polarizer 960 has a second polarization direction
orthogonal to the first polarization direction, the first
quarter-wave plate 970 has a first optical axis direction, the
second quarter-wave plate 980 has a second optical axis direction,
and both the first optical axis direction and the second optical
axis direction are disposed within a range formed by the first
polarization direction and the second polarization direction.
[0074] In one embodiment, the first polarization direction is along
a vertical direction, the second polarization direction is along a
horizontal direction, and both the first optical axis direction and
the second optical axis direction are along a 45 degrees direction
in-between the first polarization direction and the second
polarization direction. When in operation, the wide band light beam
10 of the wide band light source apparatus 100 will be polarized in
a vertical direction after passing through the first polarizer 950,
and then split by the beam splitter 300 into a first branch and a
second branch. The first branch will pass through the second
quarter-wave plate 980 of the reference arm unit 800 to become a
first clockwise circularly polarized light beam, which then becomes
a first counter clockwise circularly polarized light beam after
reflected by the reflective mirror 830, and the first counter
clockwise circularly polarized light beam will become polarized in
the horizontal direction after it passes through the second
quarter-wave plate 980. The second branch will pass through the
first quarter-wave plate 970 disposed between the first objective
lens 500 and the first dichroic splitter 400 to become a second
clockwise circularly polarized light beam, which then becomes a
second counter clockwise circularly polarized light beam after
reflected by the sample carrier unit 600, and the second counter
clockwise circularly polarized light beam will become polarized in
the horizontal direction after it passes through the first
quarter-wave plate 970. As the second polarizer 960 in front of the
two-dimensional monochrome sensing device 940 will only allow
horizontally polarized light of the wide band light beam 10 to pass
through, therefore, reflect light due to an antireflection film of
the beam splitter 300 can be effectively blocked to further enhance
an optical interference effect and promote an imaging
performance.
[0075] Please refer to FIG. 7, which illustrates a block diagram of
an optical sectioning apparatus using optical interference
microscopy and fluorescence microscopy according to still another
embodiment of the present invention.
[0076] As illustrated in FIG. 7, the reference arm unit 800 further
includes an axial motion platform 840 and the sample carrier unit
600 further includes a 3-D motion platform 630. In addition, an
information processing apparatus (not shown in this figure) can be
used to perform an image processing process.
[0077] By using the axial motion platform 840 to move the second
objective lens 820 and the reflective mirror 830, adjusting the
optical path delay unit 500, and using the 3-D motion platform 630
to move the sample 610 stained with a fluorescent dye, the
information processing apparatus (not shown in this figure) can
derive a 3-D image (not shown in this figure) of the sample
accordingly. Since the formation process of a 3-D image is known in
prior art, it will not be addressed further.
[0078] Due to the novel schemes disclosed above, the present
invention possesses the advantages as follows:
[0079] 1. The optical sectioning apparatus of the present invention
has a first dichroic splitter disposed between a beam splitter and
a first objective lens to block a short-wavelength light beam and
thereby increase a relative strength of a fluorescent signal,
shorten an exposure time, and speed up taking images.
[0080] 2. The optical sectioning apparatus of the present invention
ensures that when a fluorescent light beam emitted from a sample
passes through a first dichroic splitter, since the first dichroic
splitter will filter out a short-wavelength light beam, the
fluorescent light beam will have a good relative strength when it
passes through the first dichroic splitter, thereby shortening an
exposure time and speeding up taking images.
[0081] 3. The optical sectioning apparatus of the present invention
has a long wavelength pass filter installed in a sensor unit to
further filter out a short-wavelength light beam to increase the
relative strength of a fluorescent signal, and thereby shortening
an exposure time and speeding up taking images.
[0082] While the invention has been described by way of example and
in terms of preferred embodiments, it is to be understood that the
invention is not limited thereto. On the contrary, it is intended
to cover various modifications and similar arrangements and
procedures, and the scope of the appended claims therefore should
be accorded the broadest interpretation so as to encompass all such
modifications and similar arrangements and procedures.
[0083] In summation of the above description, the present invention
herein enhances the performance over the conventional structure and
further complies with the patent application requirements and is
submitted to the Patent and Trademark Office for review and
granting of the commensurate patent rights.
* * * * *