CN115144951B - Differential phase contrast imaging device based on fiber array illumination - Google Patents
Differential phase contrast imaging device based on fiber array illumination Download PDFInfo
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- 238000003384 imaging method Methods 0.000 title claims abstract description 107
- 238000005286 illumination Methods 0.000 title claims abstract description 61
- 239000000835 fiber Substances 0.000 title claims abstract description 23
- 239000013307 optical fiber Substances 0.000 claims abstract description 37
- 238000003491 array Methods 0.000 claims abstract description 17
- 230000003287 optical effect Effects 0.000 abstract description 9
- 238000005516 engineering process Methods 0.000 abstract description 8
- 239000000523 sample Substances 0.000 description 9
- 238000002135 phase contrast microscopy Methods 0.000 description 7
- 238000010586 diagram Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000012472 biological sample Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000010287 polarization Effects 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
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- G—PHYSICS
- G02—OPTICS
- 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 fiber array illumination, which comprises: the optical fiber arrays and the imaging detectors 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 array of sub-light sources is used to generate imaging light waves forming at least one fan-shaped or one circular illumination pattern; the circles and sectors are concentric and of equal radius. According to the invention, the optical fiber array is used as an illumination light source, the number of the light sources is more than that of the light sources of the LEDs under the same illumination area, and the energy of the optical fibers is stronger, so that the number and the capacity of the light sources entering the optical system are increased under the same imaging distance and the same imaging lens, and the imaging distance, the object field and the numerical aperture of the imaging objective lens of the system can be reduced under the same light source condition, 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 a differential phase contrast imaging technology is expanded.
Description
Technical Field
The invention belongs to the technical field of biological sample observation imaging, and particularly relates to a differential phase contrast imaging device based on fiber array illumination.
Background
In the current biological sample observation imaging technology, most of vital biological structures such as cells are almost transparent, so light can directly penetrate through the biological structure samples (almost without absorption loss), namely, the transmitted light has almost no change in amplitude, and at the moment, the traditional optical microscope can not image the appearance of the samples. However, when light passes through the sample, the transmitted light at different locations of the sample will have different transmitted optical paths due to changes in the morphology, thickness, etc. of the sample, the optical path changes resulting in changes in the phase distribution of the transmitted light. The phase contrast microscopic imaging technology utilizes the point, and the appearance observation of the transparent sample is realized by extracting the phase information of the sample.
Currently, three methods for microscopic imaging based on phase information extraction are as follows:
(1) The T.E. Gureyev and K.A. Nugent implement phase information extraction of the target sample by performing a Fourier transform on the intensity propagation equation (Transport of intensity equation, TIE).
(2) Nomarski G constructs a differential interference imaging (DIC) microscope by using a prism that introduces a spatial and angular shift to the orthogonal polarized light, achieving three-dimensional pseudo-stereoscopic images of the target sample.
(3) On the basis of DIC, a new differential Contrast imaging technique, differential phase Contrast imaging technique (DIFFERENTIAL PHASE Contrast, DPC), has been developed, which can obtain imaging results similar to DIC.
However, since DIC exploits the phenomenon of birefringence of polarized light in crystals, anisotropic crystals must be used during imaging, which is not applicable to some samples. While DPC can make up for the shortage of DIC, in DPC imaging technology, the number of light sources in the illumination pattern and the intensity of light all have a great influence on the final high-resolution reconstruction. DPC itself requires that a sufficiently high resolution image be obtained, which requires that as many LEDs (LIGHT EMITTING diodes) as possible be included in the semicircular illumination pattern, the greater the number of light sources, the greater the intensity of the light, and the better the quality of the reconstructed high resolution image. The existing DPC imaging technology uses LED arrays as light sources, and the distance between the mature 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 is increased so that more LED light sources enter the aperture of the optical system, thereby increasing the imaging distance of the system, the object field of view and the numerical aperture of the imaging objective lens, and increasing the volume and the cost of the system.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a differential phase contrast imaging device based on fiber array illumination. The technical problems to be solved by the invention are realized by the following technical scheme:
a differential phase contrast imaging device based on fiber array illumination, comprising: the optical fiber arrays and the imaging detectors 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-arrays of light sources 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 one embodiment of the present invention, the 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 light passing aperture.
In one embodiment of the invention, at least two of the fan-shaped illumination patterns are spaced apart or adjacent when the array of sub-light sources produces imaging light waves for forming a plurality of fan-shaped illumination patterns.
In one embodiment of the invention, the plurality of fan-shaped illumination patterns are equal in area and equal in central angle to 15 degrees.
In one embodiment of the invention, the areas of the at least two fan-shaped illumination patterns are equal.
In one embodiment of the invention, the array of sub-light sources produces a first imaging light wave and a second imaging light wave, respectively;
When the optical fiber array generates the first imaging light wave, the imaging detector acquires a frame of first image I 1 of the target object;
When the optical fiber array generates the second imaging light wave, the imaging detector acquires a frame of second image I 2 of the target object;
generating a target image of differential phase contrast imaging according to a formula;
Wherein I DPC denotes a 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:
According to the invention, the optical fiber array is used as an illumination light source, the number of the light sources is more than that of the light sources of the LEDs under the same illumination area, and the energy of the optical fibers is stronger, so that the number and the capacity of the light sources entering the optical system are increased under the same imaging distance and the same imaging lens, and the imaging distance, the object field and the numerical aperture of the imaging objective lens of the system can be reduced under the same light source condition, 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 a 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 structural diagram of a differential phase contrast imaging device based on fiber array illumination according to an embodiment of the present invention;
FIG. 2 is a schematic distribution diagram of an optical fiber array according to an embodiment of the present invention;
Fig. 3 is a schematic diagram of an illumination pattern of an imaging light wave with an optical fiber array illuminated 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 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: an optical fiber array 1 and an imaging detector 4 disposed on both sides of the stage 2. The stage 2 is used for placing a target object 3, typically a transparent phase object of the target object 3.
The fiber array 1 is connected to 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 stage 2, which 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 part of a circle. The object 3 is illuminated by the array of sub-light sources 5 and imaged in the imaging detector 4.
In this embodiment, the optical fiber array 1 is used as an illumination light source, the number of light sources is larger than that of the LEDs under the same illumination area, so as to provide illumination with larger density, and the number and capability of light sources entering the optical system are increased under the same imaging distance and imaging objective lens, so that a reconstructed image with higher resolution can be obtained. The use of the fiber array 1 as a light source can enable the differential phase contrast imaging device to still obtain a high-resolution reconstructed image under the condition of a low-cost low-numerical aperture imaging objective lens with a small imaging distance. The optical fiber array 1 for generating the fan-shaped distributed imaging light source can provide various illumination patterns based on different shapes of the fan, so that the imaging device of the embodiment still has universality under different imaging conditions, and the application scene of differential phase contrast imaging technology is expanded.
The imaging device of the embodiment improves stability, reduces cost, breaks through the limit of the prior DPC technology on imaging distance and numerical aperture of the imaging objective lens, and can still obtain high-resolution reconstructed images under the condition of low-numerical aperture imaging objective lens.
Wherein the spacing between the optical fibers in the optical fiber array 1 is less than or equal to 125 microns, which is much less than the spacing of 4 millimeters of the LED array.
Further, a plurality of optical fibers corresponding to the sub-light source array 5 are coupled as a switch. The optical fiber array 1 may be arranged in any shape as required, 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 optical fibers corresponding to each sub-light source array 5 are coupled into an independent switch, and all the optical fibers except the circular light emitting area, that is, all the optical fibers except the sub-light source arrays 5, are coupled into an independent switch, and the imaging light waves for forming the fan-shaped or circular illumination pattern can be formed through the switches corresponding to the switches. When an imaging light wave forming a circular illumination pattern is required, all sub-light source arrays 5 are all lit up on.
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 object 3, the distance from the target object 3 to the lens of the imaging detector 4, and the radius of the light passing aperture. Specifically, the radius of the light source= (distance of the optical fiber array 1 to the target object 3/distance of the target object 3 to the lens of the imaging detector 4) ×the radius of the light-transmitting 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. Each time the optical fiber array 1 is illuminated to generate an imaging light wave, any sub-light source array 5, that is, a light source generating any number of fan-shaped illumination patterns, may be illuminated. For example, the illumination pattern of the imaging light wave is a large sector with a large area formed by a plurality of sectors, or the illumination pattern of the imaging light wave is a plurality of sectors with intervals, and the illumination pattern of the imaging light wave is a plurality of adjacent sub-light source arrays 5 and a plurality of non-adjacent (intervals) sub-light source arrays 5, or one sub-light source array 5, etc. are illuminated at once, and any sub-light source array 5 can be illuminated at once according to imaging requirements.
Preferably, the areas of the fan-shaped illumination patterns corresponding to each sub-light source array 5 are equal, and the central angles are equal to 15 degrees, and the fan shape of the 15-degree central angle is convenient for forming the common angle condition.
Of course, the areas of the fan-shaped illumination patterns corresponding to each sub-light source array 5 may be all unequal, or at least two of the fan-shaped illumination patterns may be equal, and the areas of the remaining fan-shaped illumination patterns may be all unequal.
Further, it should be noted that, the phase contrast microscope is mainly used to improve the resolution of transparent phase objects, such as undyed cells, under the optical microscope, and mainly includes zernike phase contrast microscopy and nomads differential interference phase contrast microscopy. Zernike phase contrast microscopy generally requires the introduction of a phase ring within the aperture plane of the condenser lens by means of a phase difference objective lens, so that the conjugate area is adapted to the annular phase plane within the phase difference objective lens, and the resolution of a transparent phase object under an optical microscope is greatly improved by changing the spectral phase of the object light wave by the principle of such spatial filtering. The Normaski differential interference phase contrast microscopy is based on the polarization beam splitting interference principle, and can convert the phase gradient of a sample into intensity difference to be reflected, so that a pseudo three-dimensional relief image is formed, and the pseudo three-dimensional relief image has strong stereoscopic impression. As typical two phase contrast microscopy methods, phase contrast microscopy and differential interference phase contrast microscopy can realize visualization of phase objects such as cells without staining the sample, and have become almost the common method for all biological microscopes.
The invention adopts a differential phase contrast method for imaging, the imaging principle schematic diagram is shown in figure 1, parameters such as imaging distance and the like are calculated, a light path is calibrated, an optical fiber control end is controlled to enable a light source array 5 of an optical fiber array 1 to be lightened twice, a first imaging light wave and a second imaging light wave are respectively generated, namely, light sources with two illumination patterns are respectively generated, an imaging detector 4 respectively acquires one frame of image each time, differential phase contrast imaging can be realized after the two frames of images are processed by a computer, and a target image is generated.
Specifically, when the optical fiber array 1 generates a first imaging light wave, the imaging detector 4 acquires a frame of first image I 1 of the target object 3;
When the optical fiber array 1 generates a second imaging light wave, the imaging detector 4 acquires a frame of second image I 2 of the target object 3;
generating a target image of differential phase contrast imaging according to a formula;
Wherein I DPC denotes a target image.
In the description of the present invention, it should 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", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present invention, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.
In the present invention, unless explicitly specified and limited otherwise, the terms "mounted," "connected," "secured," and the like are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communicated with the inside of two elements or the interaction relationship of the two elements. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art according to the specific circumstances.
In the present invention, unless expressly stated or limited otherwise, a first feature "above" or "below" a second feature may include both the first and second features being in direct contact, as well as the first and second features not being in direct contact but being in contact with each other through additional features therebetween. Moreover, a first feature being "above," "over" and "on" a second feature includes the first feature being directly above and obliquely above the second feature, or simply indicating that the first feature is higher in level than the second feature. The first feature being "under", "below" and "beneath" the second feature includes the first feature being directly under and obliquely below the second feature, or simply means that the first feature is less level than the second feature.
In the description of the present specification, a description referring to terms "one embodiment," "some embodiments," "examples," "specific examples," or "some examples," etc., 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 present invention. In this specification, schematic representations of the above terms are not necessarily directed 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. Further, one skilled in the art can engage and combine the different embodiments or examples described in this specification.
The foregoing is a further detailed description of the invention in connection with the preferred embodiments, and it is not intended that the invention be limited to the specific embodiments described. It will be apparent to those skilled in the art that several simple deductions or substitutions may be made without departing from the spirit of the invention, and these should be considered to be within the scope of the invention.
Claims (8)
1. A differential phase contrast imaging device based on fiber array illumination, comprising: the optical fiber arrays (1) and the imaging detectors (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-arrays of light sources (5) for generating imaging light waves incident towards said stage (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. A differential phase contrast imaging device based on fiber array illumination according to claim 1, characterized in that the plurality of fibers corresponding to the sub-light source array (5) are coupled as a switch.
3. A differential phase contrast imaging device based on fiber array illumination according to claim 1, characterized in that the light source radius of the fan-shaped illumination pattern is determined from the distance of the fiber array (1) to the target object (3), the distance of the target object (3) to the lens of the imaging detector (4) and the radius of the light passing aperture.
4. A differential phase contrast imaging device based on fiber array illumination according to claim 1, characterized in that at least two fan-shaped illumination patterns are spaced apart or adjacent when the array of sub-light sources (5) generates imaging light waves for forming a plurality of fan-shaped illumination patterns.
5. A differential phase contrast imaging apparatus based on fiber array illumination according to claim 1, wherein the plurality of fan-shaped illumination patterns are equal in area and equal in central angle to 15 degrees.
6. A differential phase contrast imaging apparatus based on fiber array illumination according to claim 1, wherein the areas of at least two fan-shaped illumination patterns are equal.
7. A differential phase contrast imaging device based on fiber array illumination according to claim 1, characterized in that the array of sub-light sources (5) generates a first imaging light wave and a second imaging light wave, respectively;
When the optical fiber array (1) generates the first imaging light wave, the imaging detector (4) acquires a frame of first image I 1 of the target object (3);
When the optical fiber array (1) generates the second imaging light wave, the imaging detector (4) acquires a frame of second image I 2 of the target object (3);
generating a target image of differential phase contrast imaging according to a formula;
Wherein I DPC denotes a target image.
8. A differential phase contrast imaging device based on fiber array illumination according to claim 1, characterized in that the spacing between the fibers in the fiber array (1) is less than or equal to 125 micrometers.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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| US20020015952A1 (en) * | 1999-07-30 | 2002-02-07 | Anderson Norman G. | Microarrays and their manufacture by slicing |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (1)
| Title |
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| Holographic pixel super-resolution in portable lensless on-chip microscopy using a fiber-optic array;Waheb Bishara et.al;《Lab on a Chip》;全文 * |
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