CN111366537A - Dark field polarization extinction imaging device and method for biological tissue detection - Google Patents
Dark field polarization extinction imaging device and method for biological tissue detection Download PDFInfo
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- CN111366537A CN111366537A CN202010172664.1A CN202010172664A CN111366537A CN 111366537 A CN111366537 A CN 111366537A CN 202010172664 A CN202010172664 A CN 202010172664A CN 111366537 A CN111366537 A CN 111366537A
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- 238000003384 imaging method Methods 0.000 title claims abstract description 73
- 230000010287 polarization Effects 0.000 title claims abstract description 36
- 230000008033 biological extinction Effects 0.000 title claims abstract description 33
- 238000001514 detection method Methods 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title description 6
- 210000001519 tissue Anatomy 0.000 claims abstract description 56
- 210000002615 epidermis Anatomy 0.000 claims abstract description 8
- 238000005516 engineering process Methods 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 238000005286 illumination Methods 0.000 description 8
- 101100166427 Arabidopsis thaliana CCD4 gene Proteins 0.000 description 5
- 230000003287 optical effect Effects 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000012634 optical imaging Methods 0.000 description 2
- 230000000149 penetrating effect Effects 0.000 description 2
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/21—Polarisation-affecting properties
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N2021/0106—General arrangement of respective parts
- G01N2021/0112—Apparatus in one mechanical, optical or electronic block
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/21—Polarisation-affecting properties
- G01N2021/217—Measuring depolarisation or comparing polarised and depolarised parts of light
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Abstract
The invention discloses a dark field polarization extinction imaging device for biological tissue detection, which comprises an annular light source, a polarizing film A, a conical reflector, a CCD/CMOS sensor, an imaging lens and a polarizing film B, wherein annular parallel light emitted by the annular light source penetrates through the polarizing film A and then is incident to the inner surface of the conical reflector, and the conical reflector reflects light to the surface of a sample; the positions of the polaroid B, the imaging lens and the CCD/CMOS sensor are sequentially distributed from front to back, the central lines of the polaroid B, the imaging lens and the CCD/CMOS sensor are overlapped, and the reflected light and the scattered light on the surface of the sample sequentially pass through the polaroid B and the imaging lens to be imaged on the CCD/CMOS sensor. The invention filters the reflected light on the surface of the sample to obtain the high-contrast image of the superficial layer of the epidermis of the biological tissue sample. The annular parallel light is reflected to the surface of the sample through the conical reflector to form a dark field environment, so that the resolution of the polarization extinction imaging system is improved.
Description
Technical Field
The invention relates to the technical field of optics, in particular to a dark field polarization extinction imaging device for biological tissue detection.
Background
At present, most of the existing devices for imaging living biological tissues use common natural light illumination to acquire surface morphology information of the biological tissues. Chinese patent publication No. CN106901698A discloses a dual-band spectral imager based on adjustable polarization, which includes: a light source assembly for forming linearly polarized light; the beam splitting prism I reflects and focuses linearly polarized light on the biological tissue to be detected and transmits light returning from the biological tissue to be detected; further comprising an imaging system for imaging, the imaging system comprising: the imaging objective lens is used for enabling light returning from the biological tissue to be detected and passing through the beam splitter prism I to pass through and be imaged; the beam splitting prism II is arranged behind the imaging objective lens along the imaging light path and is used for splitting the imaging light path into a transmission light path and a reflection light path; the optical filter I is arranged corresponding to the transmission light path of the beam splitter prism II and is used for the light of the first specific wave band to pass through; the optical filter II is arranged corresponding to the reflection light path of the beam splitter prism II and used for the light of the second specific wave band to pass through; the imaging system also comprises a first waveband detector and a second waveband detector which are arranged in a one-to-one correspondence manner between the optical filter I and the optical filter II; the spectral imager also comprises an analyzer plate used for removing non-target light in the imaging light path, and the polarization direction of the analyzer plate is adjustable with the polarization direction of linearly polarized light formed by the light source assembly. The contrast of the surface morphology information of the biological tissue acquired by the existing device for imaging the living body of the biological tissue is not high enough, and the resolution of an imaging system is low.
Disclosure of Invention
The invention provides a dark field polarization extinction imaging device for biological tissue detection, which solves the problems that the contrast of the device for imaging a biological tissue living body in the prior art for acquiring surface morphology information of the biological tissue is not high enough, and the resolution of an imaging system is low.
The technical scheme of the invention is realized as follows:
a dark field polarization extinction imaging device for biological tissue detection comprises an annular light source, a polarizing film A, a conical surface reflector, a CCD/CMOS sensor, an imaging lens and a polarizing film B, wherein annular parallel light emitted by the annular light source penetrates through the polarizing film A and then is incident to the inner surface of the conical surface reflector, and the conical surface reflector reflects light to the surface of a sample; the positions of the polaroid B, the imaging lens and the CCD/CMOS sensor are sequentially distributed from front to back, the central lines of the polaroid B, the imaging lens and the CCD/CMOS sensor are overlapped, and the reflected light and the scattered light on the surface of the sample sequentially pass through the polaroid B and the imaging lens to be imaged on the CCD/CMOS sensor.
As a preferred embodiment of the present invention, the polarization directions of the polarizing plate a and the polarizing plate B are orthogonally arranged.
In a preferred embodiment of the present invention, the reflected light and the scattered light on the surface of the sample pass through the polarizer B, the polarizer B filters the reflected light, and the scattered light passes through the polarizer B and the imaging lens to be imaged on the CCD/CMOS sensor.
As a preferred embodiment of the present invention, the conical surface reflector is an inner conical surface reflector.
As a preferred embodiment of the present invention, the imaging lens is a variable focus imaging lens.
A dark field polarization extinction imaging method for biological tissue detection specifically comprises the following steps:
emitting oblique rays to the surface of a biological tissue sample in a dark field environment, and emitting emitted light and scattered light from the surface of the sample;
based on the polarized extinction technology, reflected light emitted from the surface of the sample is filtered, and scattered light emitted from the surface of the sample enters an imaging system to generate a high-contrast image of a superficial layer of the epidermis of the biological tissue sample.
As a preferred embodiment of the present invention, filtering reflected light emitted from a sample surface based on a polarization extinction technique specifically means that a polarizer a and a polarizer B are arranged, the polarization directions of which are orthogonal, and the polarizer a and the polarizer B are respectively arranged in the paths of incident light and emergent light on the sample surface, the reflected light and the scattered light on the sample surface pass through the polarizer B, and the polarizer B filters the reflected light.
As a preferred embodiment of the present invention, emitting oblique light to the surface of the biological tissue sample in a dark-field environment specifically means that the ring light source emits ring-shaped parallel light, which passes through the polarizer a and then is incident on the inner surface of the conical reflector, and the conical reflector reflects the light to the surface of the biological tissue sample.
The invention has the beneficial effects that: and filtering reflected light on the surface of the sample to obtain a high-contrast image of the superficial layer of the epidermis of the biological tissue sample. The annular parallel light is reflected to the surface of the sample through the conical reflector to form a dark field environment, so that the resolution of the polarization extinction imaging system is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a schematic structural diagram of one embodiment of a dark field polarization extinction imaging device for biological tissue detection according to the present invention;
FIG. 2 is a flow chart of a dark field polarization extinction imaging device for biological tissue detection according to the present invention;
FIG. 3 is a comparison of prior art and present invention imaging.
In the figure, 1-ring light source, 2-polaroid A, 3-conical reflector, 4-CCD/CMOS sensor, 5-imaging lens, 6-polaroid B, 7-sample surface.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it should be noted that the terms "vertical", "upper", "lower", "horizontal", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of describing the present invention and simplifying the description, but do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention.
In the description of the present invention, it should also be noted that, unless otherwise explicitly specified or limited, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
As shown in fig. 1, the present invention provides a dark field polarization extinction imaging device for biological tissue detection, which includes an annular light source 1, a polarizer a2, a conical reflector 3, a CCD4/CMOS sensor, an imaging lens 5 and a polarizer B6, wherein annular parallel light emitted by the annular light source 1 passes through the polarizer a2 and then is incident on the inner surface of the conical reflector 3, and the conical reflector 3 reflects light to a sample surface 7; the polaroid B6, the imaging lens 5 and the CCD4/CMOS sensor are sequentially distributed from front to back, the central lines of the polaroid B6, the imaging lens 5 and the CCD4/CMOS sensor coincide with each other, and reflected light and scattered light on the surface 7 of the sample sequentially pass through the polaroid B6 and the imaging lens 5 to be imaged on the CCD4/CMOS sensor.
When the biological tissue sample is detected, the polarized extinction imaging can filter out surface reflected light to obtain information of a shallow layer below the surface. The invention adopts oblique illumination method to block the direct light penetrating through the sample detail, and observes the sample by reflected light and diffracted light. The object morphology and structure is seen relative to the direct illumination of the sample under ordinary optical imaging. In dark field illumination, the light beam that is incident on the object from the side is diffracted or reflected to form a silhouette of the object. The profile of the sample structure seen under the dark field microscope is therefore. The invention filters the reflected light of the sample surface 7 to obtain the high-contrast image of the superficial layer of the epidermis of the biological tissue sample; and the resolution of the polarization extinction imaging system is improved.
In a preferred embodiment of the present invention, the polarization directions of the polarizer a2 and the polarizer B6 are orthogonally arranged, the reflected light and the scattered light from the sample surface 7 pass through the polarizer B6, the polarizer B6 filters the reflected light, and the scattered light passes through the polarizer B6 and the imaging lens 5 to be imaged on the CCD4/CMOS sensor. Specifically, the light incident on the biological tissue sample from the ring light source 1 via the polarizer a2 is polarized, and after being reflected by the sample surface 7, the polarized light cannot pass through the polarizer B6 and is filtered out. The light of the annular light source 1, which is incident on the biological tissue sample through the polarizer a2, is polarized, and after entering the biological tissue sample, the polarized light is scattered and depolarized, and can be imaged through the polarizer B6. The reflected light from the sample surface 7 is filtered out, which improves the contrast of the image.
As a preferred embodiment of the present invention, the conical surface reflector 3 is an inner conical surface reflector 3. After passing through the conical reflector 3, the annular parallel light is obliquely incident to the sample to form dark field illumination, so that the resolution of the imaging system can be improved.
As a preferred embodiment of the present invention, the imaging lens 5 is a variable focus imaging lens 5. The variable focus imaging lens 5 may improve the applicability of the invention.
As shown in fig. 2, the present invention further provides a dark-field polarization extinction imaging method for biological tissue detection, which specifically includes the following steps:
emitting oblique rays to the surface 7 of the biological tissue sample in a dark field environment, and emitting emitted light and scattered light from the surface 7 of the sample;
based on the polarized extinction technology, the reflected light emitted from the sample surface 7 is filtered, and the scattered light emitted from the sample surface 7 enters an imaging system to generate a high-contrast image of the superficial layer of the epidermis of the biological tissue sample.
As a preferred embodiment of the present invention, filtering the reflected light emitted from the sample surface 7 based on the polarization extinction technology specifically means that a polarizer a2 and a polarizer B6 with orthogonal polarization directions are disposed, a polarizer a2 and a polarizer B6 are respectively disposed in the path of the incident ray and the emergent ray of the sample surface 7, the reflected light and the scattered light of the sample surface 7 pass through the polarizer B6, and the polarizer B6 filters the reflected light. Specifically, the emitting of oblique light to the biological tissue sample surface 7 in the dark field environment means that the annular light source 1 emits annular parallel light, which passes through the polarizer a2 and then is incident on the inner surface of the conical reflector 3, and the conical reflector 3 reflects the light to the biological tissue sample surface 7. Specifically, the light incident on the biological tissue sample from the ring light source 1 via the polarizer a2 is polarized, and after being reflected by the sample surface 7, the polarized light cannot pass through the polarizer B6 and is filtered out. The light of the annular light source 1, which is incident on the biological tissue sample through the polarizer a2, is polarized, and after entering the biological tissue sample, the polarized light is scattered and depolarized, and can be imaged through the polarizer B6. The reflected light from the sample surface 7 is filtered out, which improves the contrast of the image.
When the biological tissue sample is detected, the polarized extinction imaging can filter out surface reflected light to obtain information of a shallow layer below the surface. The invention adopts oblique illumination method to block the direct light penetrating through the sample detail, and observes the sample by reflected light and diffracted light. The object morphology and structure is seen relative to the direct illumination of the sample under ordinary optical imaging. In dark field illumination, the light beam that is incident on the object from the side is diffracted or reflected to form a silhouette of the object. The profile of the sample structure seen under the dark field microscope is therefore. The invention filters the reflected light of the sample surface 7 to obtain the high-contrast image of the superficial layer of the epidermis of the biological tissue sample; and the resolution of the polarization extinction imaging system is improved.
As shown in fig. 3, fig. 3(a) is an image obtained by using normal natural light to illuminate and obtain surface morphology information of a biological tissue, and fig. 3(b) is an image obtained by using the dark field polarization extinction imaging device or method for biological tissue detection to obtain surface morphology information of a biological tissue, and comparing the two images, it can be seen that the details of the images are more obvious, and the later research and processing of the biological tissue can be conveniently performed according to the collected surface morphology information.
The invention has the beneficial effects that: and filtering reflected light on the surface 7 of the sample to obtain a high-contrast image of the superficial layer of the epidermis of the biological tissue sample. The annular parallel light is reflected to the surface 7 of the sample through the conical reflector 3 to form a dark field environment, so that the resolution of the polarization extinction imaging system is improved.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. A dark field polarized extinction imaging device for biological tissue detection, characterized by: the device comprises an annular light source, a polaroid A, a conical reflector, a CCD/CMOS sensor, an imaging lens and a polaroid B, wherein annular parallel light emitted by the annular light source penetrates through the polaroid A and then is incident to the inner surface of the conical reflector, and the conical reflector reflects light to the surface of a sample; the positions of the polaroid B, the imaging lens and the CCD/CMOS sensor are sequentially distributed from front to back, the central lines of the polaroid B, the imaging lens and the CCD/CMOS sensor are overlapped, and the reflected light and the scattered light on the surface of the sample sequentially pass through the polaroid B and the imaging lens to be imaged on the CCD/CMOS sensor.
2. The dark field polarization extinction imaging device for biological tissue detection according to claim 1, wherein: the polarization directions of the polaroid A and the polaroid B are orthogonally arranged.
3. The dark field polarization extinction imaging device for biological tissue detection according to claim 2, wherein: and the reflected light and the scattered light on the surface of the sample pass through the polaroid B, the polaroid B filters the reflected light, and the scattered light passes through the polaroid B and the imaging lens to be imaged on the CCD/CMOS sensor.
4. The dark field polarization extinction imaging device for biological tissue detection according to claim 1, wherein: the conical surface reflector is an inner conical surface reflector.
5. The dark field polarization extinction imaging device for biological tissue detection according to claim 1, wherein: the imaging lens is a variable focus imaging lens.
6. A dark field polarization extinction imaging method for biological tissue detection is characterized by comprising the following steps:
emitting oblique rays to the surface of a biological tissue sample in a dark field environment, and emitting emitted light and scattered light from the surface of the sample;
based on the polarized extinction technology, reflected light emitted from the surface of the sample is filtered, and scattered light emitted from the surface of the sample enters an imaging system to generate a high-contrast image of a superficial layer of the epidermis of the biological tissue sample.
7. The dark field polarization extinction imaging method for biological tissue detection according to claim 6, wherein: the filtering of the reflected light emitted from the sample surface based on the polarization extinction technology specifically means that a polarizing film a and a polarizing film B with orthogonal polarization directions are arranged, the polarizing film a and the polarizing film B are respectively arranged in the paths of the incident light and the emergent light on the sample surface, the reflected light and the scattered light on the sample surface pass through the polarizing film B, and the polarizing film B filters the reflected light.
8. The dark field polarization extinction imaging method for biological tissue detection according to claim 7, wherein: emitting oblique rays to the surface of the biological tissue sample in a dark field environment specifically means that an annular light source emits annular parallel light, the annular parallel light passes through the polarizing film A and then enters the inner surface of the conical reflector, and the conical reflector reflects the light to the surface of the biological tissue sample.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112964726A (en) * | 2021-02-05 | 2021-06-15 | 上海御微半导体技术有限公司 | Defect detection device and method |
| CN113406842A (en) * | 2021-06-23 | 2021-09-17 | 湘潭大学 | Ultramicro stacked dark field photography system and method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112964726A (en) * | 2021-02-05 | 2021-06-15 | 上海御微半导体技术有限公司 | Defect detection device and method |
| CN113406842A (en) * | 2021-06-23 | 2021-09-17 | 湘潭大学 | Ultramicro stacked dark field photography system and method |
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