CN115144951A - Differential phase contrast imaging device based on optical fiber array illumination - Google Patents
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- 238000003384 imaging method Methods 0.000 title claims abstract description 105
- 238000005286 illumination Methods 0.000 title claims abstract description 61
- 239000013307 optical fiber Substances 0.000 title claims abstract description 49
- 238000003491 array Methods 0.000 claims abstract description 15
- 239000000835 fiber Substances 0.000 claims description 11
- 230000003287 optical effect Effects 0.000 abstract description 9
- 238000005516 engineering process Methods 0.000 abstract description 7
- 239000000523 sample Substances 0.000 description 11
- 238000000034 method Methods 0.000 description 6
- 238000010586 diagram Methods 0.000 description 4
- 238000002135 phase contrast microscopy Methods 0.000 description 4
- 239000012472 biological sample Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
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- G—PHYSICS
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- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B6/00—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings
- G02B6/0001—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings specially adapted for lighting devices or systems
- G02B6/0005—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings specially adapted for lighting devices or systems the light guides being of the fibre type
- G02B6/0008—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings specially adapted for lighting devices or systems the light guides being of the fibre type the light being emitted at the end of the fibre
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/361—Optical details, e.g. image relay to the camera or image sensor
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Abstract
The invention discloses a differential phase contrast imaging device based on optical fiber array illumination, which comprises: the optical fiber array and the imaging detector are arranged on two sides of the objective table; the optical fiber array is connected with the coupler to form a plurality of sub light source arrays; the sub light source array is used for generating imaging light waves forming at least one fan-shaped or one circular illumination pattern; the circle and the sector are concentric and have equal radii. The optical fiber array is used as the illumination light source, the number of the light sources is more than that of the light sources of the LED under the same illumination area, and the energy of the optical fiber is stronger, so that the number and the capability of the light sources entering the optical system are increased under the condition of the same imaging distance and the same imaging lens, and the imaging distance, the object space view field and the numerical aperture of the imaging objective lens of the system can be reduced under the condition of the same light source, so that the volume of the imaging device is reduced, the cost is reduced, the resolution of a reconstructed target image is improved, and the application scene of the differential phase contrast imaging technology is expanded.
Description
Technical Field
The invention belongs to the technical field of observation and imaging of biological samples, and particularly relates to a differential phase contrast imaging device based on optical fiber array illumination.
Background
In the current biological sample observation and imaging technology, most of the vital biological structures such as cells are almost transparent, so that light can directly penetrate through the biological structure sample (almost without absorption loss), namely, the transmitted light almost has no change on amplitude, and at the moment, the traditional optical microscope cannot image the appearance of the sample. However, when light passes through the sample, due to changes in the morphology, thickness, etc. of the sample, the transmitted light at different positions of the sample will have different transmitted optical paths, and the optical path changes result in changes in the phase distribution of the transmitted light. The phase contrast microscopic imaging technology utilizes the point, and realizes the appearance observation of the transparent sample by extracting the phase information of the sample.
At present, there are three methods for microscopic imaging based on phase information extraction:
(1) T.e.gureyev and k.a.nument perform fourier transform on an intensity propagation equation (TIE), thereby extracting phase information of a target sample.
(2) Nomarski G constructs a Differential interference imaging (DIC) microscope by using a prism that can introduce a spatial displacement and an angular shift to orthogonally polarized light, implementing a three-dimensional pseudo-stereoscopic image of a target sample.
(3) Based on DIC, a new Differential Contrast imaging technique is developed, namely Differential Phase Contrast imaging (DPC), which can achieve imaging results similar to DIC.
However, since DIC utilizes the birefringence phenomenon of polarized light in a crystal, an anisotropic crystal must be used in the imaging process, which is not suitable for some samples. Although the DPC can make up for the deficiency of DIC, in the DPC imaging technique, the number of light sources in the illumination pattern and the intensity of the light have a great influence on the final high resolution reconstruction. The DPC itself needs to obtain a high resolution image, and it requires that as many LEDs (Light Emitting diodes) as possible be included in the semicircular illumination pattern, and the higher the number of Light sources, the stronger the Light intensity, and the better the quality of the reconstructed high resolution image. The existing DPC imaging technology uses an LED array as a light source, and the spacing between LED arrays in the market is 4 mm, so in order to improve the resolution, an expensive high-numerical-aperture imaging lens must be used or the imaging distance must be increased to make more LED light sources enter the aperture of an optical system, thereby increasing the imaging distance, the object field and the numerical aperture of the imaging objective lens of the system, and increasing the volume and the cost of the system.
Disclosure of Invention
In order to solve the above problems in the prior art, the present invention provides a differential phase contrast imaging device based on fiber array illumination. The technical problem to be solved by the invention is realized by the following technical scheme:
a differential phase contrast imaging device based on fiber array illumination, comprising: the optical fiber array and the imaging detector are arranged on two sides of the objective table;
the optical fiber array is connected with the coupler to form a plurality of sub light source arrays;
a plurality of said sub-light source arrays for generating imaging light waves incident toward said stage for forming at least one fan-shaped or one circular illumination pattern;
the circular illumination pattern and the fan-shaped illumination pattern are concentric and have equal radii.
In an embodiment of the present invention, a plurality of optical fibers corresponding to the sub-light source array are coupled as a switch.
In one embodiment of the invention, the light source radius of the fan-shaped illumination pattern is determined according to the distance from the fiber array to the target, the distance from the target to the lens of the imaging detector, and the radius of the clear aperture.
In one embodiment of the invention, at least two of the fan-shaped illumination patterns are spaced apart or adjacent when the sub-light source array generates imaging light waves for forming the plurality of fan-shaped illumination patterns.
In one embodiment of the present invention, the plurality of fan shaped illumination patterns are equal in area and have central angles equal to 15 degrees.
In one embodiment of the invention, the areas of at least two of the fan-shaped illumination patterns are equal.
In one embodiment of the present invention, the array of sub-light sources generates first and second imaging light waves, respectively;
when the optical fiber array generates the first imaging light wave, the imaging detector acquires a frame of first image I of a target object 1 ；
When the optical fiber array generates the second imaging light wave, the imaging detector acquires a frame of second image I of the target object 2 ；
Generating a target image of differential phase contrast imaging according to a formula;
wherein, I DPC Representing the target image.
In one embodiment of the invention, the spacing between the optical fibers in the array of optical fibers is less than or equal to 125 microns.
The invention has the beneficial effects that:
the optical fiber array is used as the illumination light source, the number of the light sources is more than that of the LED light sources under the same illumination area, and the energy of the optical fiber is stronger, so that the number and the capacity of the light sources entering the optical system are increased under the condition of the same imaging distance and the same imaging lens, and the imaging distance, the object space view field and the numerical aperture of the imaging objective lens of the system can be reduced under the condition of the same light source, so that the volume of the imaging device is reduced, the cost is reduced, the resolution of a reconstructed target image is improved, and the application scene of the differential phase contrast imaging technology is expanded.
The present invention will be described in further detail with reference to the accompanying drawings and examples.
Drawings
FIG. 1 is a schematic diagram of an embodiment of the present invention for providing an optical fiber-based optical fiber the structure schematic diagram of the differential phase contrast imaging device of the array illumination;
FIG. 2 is a schematic diagram of a distribution of an optical fiber array provided by an embodiment of the present invention;
fig. 3 is a schematic view of an illumination pattern of an imaging light wave when an optical fiber array is lit once according to an embodiment of the present invention.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention are not limited thereto.
As shown in fig. 1 and 2, a differential phase contrast imaging device based on fiber array illumination includes: a fiber array 1 and an imaging detector 4 arranged at both sides of the stage 2. The stage 2 is used for placing an object 3, which object 3 is typically a transparent phase object.
The optical fiber array 1 is connected with a coupler to form a plurality of sub light source arrays 5.
A plurality of sub-light source arrays 5 for generating imaging light waves incident towards the object table 2, which imaging light waves can be used to form at least one fan-shaped or one circular illumination pattern; the circular and fan-shaped illumination patterns are concentric and of equal radius, the fan being a portion of a circle. The object 3 is imaged in the imaging detector 4 after being irradiated by the sub light source array 5.
In this embodiment, the optical fiber array 1 is used as an illumination light source, the number of light sources is greater than that of LEDs in the same illumination area, and illumination with a greater density is provided, and the number and capability of light sources entering the optical system are increased in the same imaging distance and imaging objective lens, so that a reconstructed image with a higher resolution can be obtained. Meanwhile, the optical fiber array 1 is used as a light source, so that the differential phase contrast imaging device can still obtain a high-resolution reconstructed image under the conditions of a small imaging distance and a low-cost low-numerical-aperture imaging objective lens. And the optical fiber array 1 for generating the fan-shaped distributed imaging light source can provide various illumination patterns based on different fan shapes, so that the imaging device of the embodiment still has universality under different imaging conditions, and the application scene of the differential phase contrast imaging technology is expanded.
The imaging device of the embodiment improves the stability, reduces the cost, breaks through the limitation of the existing DPC technology on the imaging distance and the numerical aperture of the imaging objective lens, and can still obtain a high-resolution reconstructed image under the condition of the low numerical aperture imaging objective lens.
The spacing between the optical fibers in the optical fiber array 1 is smaller than or equal to 125 micrometers, and is far smaller than the spacing of the LED array by 4 millimeters.
Further, a plurality of optical fibers corresponding to the sub-light source array 5 are coupled as a switch. The optical fiber array 1 can be set to any shape according to requirements, and the light emitting area of the optical fiber array 1 is larger than the light emitting area of the imaging light wave of the circular illumination pattern formed by the plurality of sub light source arrays 5. The plurality of optical fibers corresponding to each sub-light source array 5 are coupled to form an independent switch, all the optical fibers except the circular light emitting area, that is, all the optical fibers except all the sub-light source arrays 5 are coupled to form an independent switch, and the imaging light waves for forming the fan-shaped or circular illumination pattern can be formed by the switches corresponding to the switches. When the imaging light wave forming the circular illumination pattern is required, all the sub light source arrays 5 are turned on all by lighting.
Specifically, the radius of the light source of the fan-shaped illumination pattern is determined according to the distance from the optical fiber array 1 to the target 3, the distance from the target 3 to the lens of the imaging detector 4, and the radius of the clear aperture. Specifically, the radius of the light source = (distance from the optical fiber array 1 to the object 3/distance from the object 3 to the lens of the imaging detector 4) × the radius of the clear aperture.
Further, as shown in fig. 3, a hatched area in the drawing indicates a light-emitting area, and an unshaded area indicates no light emission. When the light source array 5 generates imaging light waves for forming a plurality of fan-shaped illumination patterns, at least two fan-shaped illumination patterns are spaced apart or adjacent to each other. Every time the optical fiber array 1 is lighted to generate imaging light waves, any sub light source array 5, namely, a light source generating any fan-shaped illumination patterns, can be lighted. For example, a plurality of adjacent sub-light source arrays 5 are simultaneously lighted at a time, the illumination pattern of the imaging light wave is a large fan shape with a large area formed by a plurality of fan shapes, or a plurality of non-adjacent (spaced) sub-light source arrays 5 are simultaneously lighted at a time, the illumination pattern of the imaging light wave is a plurality of spaced fan shapes, or a plurality of adjacent sub-light source arrays 5 and a plurality of non-adjacent (spaced) sub-light source arrays 5 are simultaneously lighted at a time, or one sub-light source array 5 is lighted at a time, and the like, and according to the imaging requirements, any sub-light source array 5 can be simultaneously lighted at a time.
Preferably, the areas of the fan-shaped illumination patterns corresponding to each sub-light source array 5 are all equal, and the central angles of the fan-shaped illumination patterns are all equal to 15 degrees, and the fan-shaped illumination patterns with the central angles of 15 degrees are convenient for forming a common angle condition.
Of course, the areas of the fan-shaped illumination patterns corresponding to each sub-light source array 5 may also be different, or at least two of the fan-shaped illumination patterns have the same area, and the areas of the remaining fan-shaped illumination patterns are different.
Further, it should be noted that the phase contrast microscope is mainly used to improve the resolution of transparent phase objects, such as unstained cells, under the optical microscope, and mainly includes zernike phase contrast microscope and nomas differential interference phase contrast microscope. Zernike phase contrast microscopy usually needs to use a phase difference objective lens, a phase ring is introduced into an aperture plane of a condenser lens, so that a conjugate area is adapted to an annular phase plane in the phase difference objective lens, and the frequency spectrum phase of an object light is changed through the spatial filtering principle, so that the distinguishability of a transparent phase object under an optical microscope is greatly improved. The Nomesis differential interference phase contrast microscope is based on the polarization light splitting interference principle, can convert the phase gradient of a sample into intensity difference to be reflected, forms a pseudo three-dimensional relief image, and has strong stereoscopic impression. As two typical phase-contrast microscopy methods, phase-contrast microscopy and differential interference phase-contrast microscopy can visualize phase objects such as cells without staining the sample, and have become common methods for all biological microscopes.
The invention adopts a differential phase contrast method for imaging, the imaging principle schematic diagram of the method is shown in figure 1, parameters such as imaging distance and the like are calculated, a light path is calibrated, the control end of an optical fiber is controlled to light a light source array 5 of the optical fiber array 1 twice, a first imaging light wave and a second imaging light wave are respectively generated, namely, the light sources are respectively used for generating two illumination patterns, an imaging detector 4 respectively collects a frame of image each time, and the differential phase contrast imaging can be realized after the two frames of images are processed by a computer to generate a target image.
Specifically, when the optical fiber array 1 generates the first imaging light wave, the imaging detector 4 collects a frame of the first image I of the target object 3 1 ；
When the optical fiber array 1 generates the second imaging light wave, the imaging detector 4 collects a frame of second image I of the target 3 2 ；
Generating a target image of differential phase contrast imaging according to a formula;
wherein, I DPC Representing the target image.
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", and the like, indicate orientations and positional relationships based on those shown in the drawings, and are used only for convenience of description and simplicity of description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be considered as limiting the present invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or to implicitly indicate the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or may be connected through the use of two elements or the interaction of two elements. The specific meanings of the above terms in the present invention can be understood according to specific situations by those of ordinary skill in the art.
In the present invention, unless expressly stated or limited otherwise, the recitation of a first feature "on" or "under" a second feature may include the recitation of the first and second features being in direct contact, and may also include the recitation that the first and second features are not in direct contact, but are in contact via another feature between them. Also, the first feature "on," "above" and "over" the second feature may include the first feature being directly above and obliquely above the second feature, or simply indicating that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature includes the first feature being directly under and obliquely below the second feature, or simply meaning that the first feature is at a lesser elevation than the second feature.
In the description of the specification, reference to the description of "one embodiment," "some embodiments," "an example," "a specific example," or "some examples" or the like means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples described in this specification can be combined and combined by those skilled in the art.
The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the invention be limited to these specific details. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.
Claims (8)
1. A differential phase contrast imaging device based on fiber array illumination, comprising: the optical fiber array (1) and the imaging detector (4) are arranged on two sides of the objective table (2);
the optical fiber array (1) is connected with a coupler to form a plurality of sub light source arrays (5);
a plurality of said sub-light source arrays (5) for generating imaging light waves incident towards said object table (2) for forming at least one fan-shaped or one circular illumination pattern;
the circular illumination pattern and the fan-shaped illumination pattern are concentric and have equal radii.
2. The differential phase contrast imaging device based on fiber array illumination according to claim 1, wherein a plurality of optical fibers corresponding to the sub light source array (5) are coupled as a switch.
3. The apparatus according to claim 1, wherein the light source radius of the fan-shaped illumination pattern is determined according to the distance from the fiber array (1) to the target (3), the distance from the target (3) to the lens of the imaging detector (4), and the radius of the clear aperture.
4. A differential phase contrast imaging apparatus based on fiber array illumination according to claim 1, wherein at least two of the fan-shaped illumination patterns are spaced or adjacent when the sub light source array (5) generates imaging light waves for forming a plurality of fan-shaped illumination patterns.
5. The differential phase-contrast imaging device based on fiber array illumination of claim 1, wherein the fan-shaped illumination patterns are equal in area and equal in central angle to 15 degrees.
6. The differential phase-contrast imaging device based on fiber array illumination of claim 1, wherein at least two fan-shaped illumination patterns are equal in area.
7. The differential phase contrast imaging device based on optical fiber array illumination according to claim 1, characterized in that the sub light source array (5) respectively generates a first imaging light wave and a second imaging light wave;
when the optical fiber array (1) generates the first imaging light wave, the imaging detector (4) collects a frame of first image I of a target object (3) 1 ；
When the optical fiber array (1) generates the second imaging light wave, the imaging detector (4) collects a frame of second image I of the target object (3) 2 ；
Generating a target image of differential phase contrast imaging according to a formula;
wherein, I DPC Representing the target image.
8. The differential phase contrast imaging device based on optical fiber array illumination according to claim 1, characterized in that the spacing between the optical fibers in the optical fiber array (1) is less than or equal to 125 μm.
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Citations (5)
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| US20020015952A1 (en) * | 1999-07-30 | 2002-02-07 | Anderson Norman G. | Microarrays and their manufacture by slicing |
| JP2015029461A (en) * | 2013-08-02 | 2015-02-16 | 克昌 藤田 | Imaging device |
| WO2016187591A1 (en) * | 2015-05-21 | 2016-11-24 | California Institute Of Technology | Laser-based fourier ptychographic imaging systems and methods |
| CN106768396A (en) * | 2016-12-23 | 2017-05-31 | 电子科技大学 | A kind of method and system based on differential contrast imaging reduction quantitative phase images |
| CN106773586A (en) * | 2017-03-15 | 2017-05-31 | 山东师范大学 | Based on single-mode fiber without lens Jones matrixes holographic measurement and imaging system |
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Patent Citations (5)
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|---|---|---|---|---|
| US20020015952A1 (en) * | 1999-07-30 | 2002-02-07 | Anderson Norman G. | Microarrays and their manufacture by slicing |
| JP2015029461A (en) * | 2013-08-02 | 2015-02-16 | 克昌 藤田 | Imaging device |
| WO2016187591A1 (en) * | 2015-05-21 | 2016-11-24 | California Institute Of Technology | Laser-based fourier ptychographic imaging systems and methods |
| CN106768396A (en) * | 2016-12-23 | 2017-05-31 | 电子科技大学 | A kind of method and system based on differential contrast imaging reduction quantitative phase images |
| CN106773586A (en) * | 2017-03-15 | 2017-05-31 | 山东师范大学 | Based on single-mode fiber without lens Jones matrixes holographic measurement and imaging system |
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| Title |
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| WAHEB BISHARA ET.AL: "Holographic pixel super-resolution in portable lensless on-chip microscopy using a fiber-optic array", 《LAB ON A CHIP》 * |
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