CN112859314A - Single-pixel scanning super-resolution phase imaging device and method - Google Patents
Single-pixel scanning super-resolution phase imaging device and method Download PDFInfo
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0036—Scanning details, e.g. scanning stages
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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
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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
- G01N21/41—Refractivity; Phase-affecting properties, e.g. optical path length
- G01N21/45—Refractivity; Phase-affecting properties, e.g. optical path length using interferometric methods; using Schlieren methods
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/0068—Optical details of the image generation arrangements using polarisation
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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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- H04—ELECTRIC COMMUNICATION TECHNIQUE
- H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
- H04N25/00—Circuitry of solid-state image sensors [SSIS]; Control thereof
- H04N25/70—SSIS architectures; Circuits associated therewith
- H04N25/71—Charge-coupled device [CCD] sensors; Charge-transfer registers specially adapted for CCD sensors
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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
- G01N21/41—Refractivity; Phase-affecting properties, e.g. optical path length
- G01N2021/4173—Phase distribution
- G01N2021/4193—Phase distribution using a PSD
Abstract
The utility model discloses a single pixel scanning super-resolution phase imaging device and method, including: the laser, the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array, the sample and the CCD image sensor are coaxially arranged in sequence along the advancing direction of the light beam, and the CCD image sensor is connected with the computer; the light beam forms a diffraction signal after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array and the sample, the diffraction signal is recorded as a diffraction image by the CCD image sensor, and the amplitude image and the phase image of the sample are obtained by the reconstruction of the diffraction image by the computer. The display of the amplitude image and the phase image of the sample is realized.
Description
Technical Field
The invention relates to the technical field of super-resolution imaging, in particular to a single-pixel scanning super-resolution phase imaging device and method.
Background
The statements in this section merely provide background information related to the present disclosure and may not necessarily constitute prior art.
As is well known, the resolution of a visible light microscope is limited to half the wavelength, which is generally considered to be 400nm to 760nm, i.e., the resolution can reach about 200 nm. The rayleigh criterion gives a way to judge the resolution, i.e. two object points can be resolved exactly when the center of the airy disk of one object point falls on the edge of the airy disk of the other object point. The rayleigh criterion is used under the condition that a light source irradiating an object is incoherent, if the light source is coherent, airy spots of two object points are coherent, and if the two object points can be distinguished, the phase relation of the two object points needs to be considered.
Two object points are considered in the Rayleigh criterion at the same time, namely the two object points are lighted at the same time, if one of the two object points is closed, only one object point exists in an object space, and only one Airy spots exists in an image space, the intensity and the phase of the Airy spots can be measured, wherein the intensity and the phase correspond to the intensity and the phase of the object point, so that the information of one pixel is obtained in the image space, and the mutual crosstalk of the Airy spots of different object points is avoided. The object space is scanned, and then a super-resolution amplitude image and a phase image of the object can be obtained.
To obtain super-resolved images, a light source with a size smaller than the limit resolution of an optical microscope is required, and the size of the light source is smaller than 400 nm. When the transparent microspheres are irradiated with parallel light, a spot having a size smaller than the diffraction limit may be generated on the other side of the microspheres, which is called nanophotonic jet. The nanometer photon jet flow can generate light spots with the diameter less than 400nm, and the light spots can be used as a light source for super-resolution imaging.
The nano photon jet flow property of the transparent medium microsphere can be applied to the fields of ultra-microscopy, optical tweezers and the like, so far, super-resolution imaging based on nano photon jet flow is limited to intensity microscopy, and phase microscopy is not realized.
Disclosure of Invention
In order to solve the above problems, the present disclosure provides a single-pixel scanning super-resolution phase imaging apparatus and method, which can obtain both an amplitude image and a phase image of a sample.
In order to achieve the purpose, the following technical scheme is adopted in the disclosure:
in a first aspect, a single-pixel scanning super-resolution phase imaging apparatus is provided, including: the laser, the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array, the sample and the CCD image sensor are coaxially arranged in sequence along the advancing direction of the light beam, and the CCD image sensor is connected with the computer;
the light beam forms a diffraction signal after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array and the sample, the diffraction signal is recorded as a diffraction image by the CCD image sensor, and the amplitude image and the phase image of the sample are obtained by the reconstruction of the diffraction image by the computer.
In a second aspect, a single-pixel scanning super-resolution phase imaging device is provided, which includes: the laser, the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first spectroscope, the first adjustable attenuation sheet, the microsphere array, the sample, the third lens, the second spectroscope and the CCD image sensor are coaxially arranged in sequence along the advancing direction of a light beam, and the first reflector, the second reflector and the second adjustable attenuation sheet are sequentially arranged along the advancing direction of the light beam split by the first spectroscope;
light beams generated by the laser form diffraction signals after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first spectroscope, the first adjustable attenuation sheet, the microsphere array, the sample, the third lens and the second spectroscope, and the diffraction signals are recorded by the CCD image sensor to obtain an amplitude image of the sample; the light beam split by the first spectroscope enters the second spectroscope after passing through the first reflector, the second reflector and the second adjustable attenuation sheet, an interference signal is generated by the second spectroscope, and the diffraction signal and the interference signal generate interference on the CCD image sensor to obtain a phase image of the sample.
In a third aspect, a single-pixel scanning super-resolution phase imaging method is provided, including:
the laser emits a light beam;
the light beam forms a diffraction signal after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array and the sample, and the diffraction signal is recorded as a diffraction image by the CCD image sensor;
the computer obtains an amplitude image and a phase image of the sample from the diffraction image reconstruction.
In a fourth aspect, a single-pixel scanning super-resolution phase imaging method is provided, including:
the laser emits a light beam;
the light beam passes through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first spectroscope, the first adjustable attenuation sheet, the microsphere array, the sample, the third lens and the second spectroscope to form a diffraction signal, and the diffraction signal is recorded by the CCD image sensor to obtain an amplitude image of the sample;
the light beam split by the first spectroscope enters the second spectroscope after passing through the first reflector, the second reflector and the second adjustable attenuation sheet, an interference signal is generated by the second spectroscope, and the diffraction signal and the interference signal generate interference on the CCD image sensor to obtain a phase image of the sample.
Compared with the prior art, the beneficial effect of this disclosure is:
1. the method can obtain the amplitude image and the phase image of the sample, and the sample is scanned by moving the sample stage to obtain the complete image of the sample, so that the technical problem that the existing super-resolution imaging is only limited to intensity microscopy and cannot realize phase microscopy is solved.
2. The disclosed light beam irradiates the microsphere array to generate a nanophotonic jet array; irradiating a sample by using a nano photon jet flow array to generate a diffraction signal, transmitting the signal to a CCD image sensor by using a lens, and recording an amplitude image of the sample; and the spectroscope divides out another beam of plane laser, the plane laser and the diffraction signal generate interference on the CCD image sensor to obtain a phase image of the sample, and the sample is moved by adopting a displacement platform to scan the sample to obtain a complete image of the sample.
3. The nanometer photon jet flow light spot of the transparent medium microsphere is used as a light source, the size of the light source is smaller than the limit resolution of a microscope, and the size and the light intensity of the light spot can be obtained according to a meter scattering theory.
4. The monolayer transparent medium microspheres are densely arranged, and the transparent film is placed between the microsphere array and the sample, so that the irradiation of nanometer light spots on the sample is ensured, and the stable placement of the transparent medium microspheres is also ensured.
5. The transparent medium microsphere is used for super-resolution microscopy to obtain an absorption contrast image, a super-resolution phase image is not reported, a CCD image sensor records the absorption contrast image of a sample under the condition of no interference light, the CCD image sensor records the phase contrast image of the sample under the condition of interference light, the step length of a nanometer displacement platform is smaller than the size of a nanometer photon jet flow light spot, the super-resolution amplitude and the phase image of the sample are obtained through two-dimensional scanning, the blank resolution between an electron microscope and an optical microscope can be made up, and the transparent medium microsphere is applied to scientific research, medical treatment and production life.
6. According to the method, the distance between adjacent light spots is larger than the minimum resolution distance of the lens, the resolution is larger than that of the lens, crosstalk between two diffraction light spots is avoided, the size of a nanometer photon jet flow light spot can be determined by selecting the size and the refractive index of the transparent medium microsphere, and therefore the resolution of an image is determined.
Advantages of additional aspects of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this application, illustrate embodiments of the application and, together with the description, serve to explain the application and are not intended to limit the application.
FIG. 1 is a schematic structural diagram of an apparatus disclosed in embodiment 1 of the present disclosure;
fig. 2 is a schematic structural diagram of an apparatus disclosed in embodiment 2 of the present disclosure.
Wherein: 1. the device comprises a laser, 2, an attenuation sheet, 3, a polarizer, 4, a first lens, 5, a diaphragm, 6, a second lens, 7, a first beam splitter, 8, a first adjustable attenuation sheet, 9, microspheres, 10, a microsphere array platform, 11, a sample, 12, a third lens, 13, a second beam splitter, 14, a first reflector, 15, a second reflector, 16, a second adjustable attenuation sheet, 17, a CCD image sensor, 18, a computer, 19 and a microsphere platform.
The specific implementation mode is as follows:
the present disclosure is further described with reference to the following drawings and examples.
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
In the present disclosure, terms such as "upper", "lower", "left", "right", "front", "rear", "vertical", "horizontal", "side", "bottom", and the like indicate orientations or positional relationships based on those shown in the drawings, and are only relational terms determined for convenience in describing structural relationships of the parts or elements of the present disclosure, and do not refer to any parts or elements of the present disclosure, and are not to be construed as limiting the present disclosure.
In the present disclosure, terms such as "fixedly connected", "connected", and the like are to be understood in a broad sense, and mean either a fixed connection or an integrally connected or detachable connection; may be directly connected or indirectly connected through an intermediate. The specific meanings of the above terms in the present disclosure can be determined on a case-by-case basis by persons skilled in the relevant art or technicians, and are not to be construed as limitations of the present disclosure.
Example 1
In the embodiment, a single-pixel scanning super-resolution phase imaging device is disclosed, and the structure of the single-pixel scanning super-resolution phase imaging device is shown in fig. 1, and the single-pixel scanning super-resolution phase imaging device comprises a laser 1, an attenuation sheet 2, a polarizer 3, a first lens 4, a diaphragm 5, a second lens 6, a first adjustable attenuation sheet 8, microspheres 9, a microsphere platform 19, a sample 11 and a CCD image sensor 17 which are coaxially arranged in sequence along the advancing direction of a light beam, wherein the CCD image sensor 17 is connected with a computer 18.
The laser 1 emits a light beam, the light beam forms a diffraction signal after passing through the attenuation sheet 2, the polarizer 3, the first lens 4, the diaphragm 5, the second lens 6, the first adjustable attenuation sheet 8, the microspheres 9 and the sample 11, the diffraction signal is recorded as a diffraction image by the CCD image sensor 17, the computer 18 reconstructs the diffraction image by adopting an iterative algorithm to obtain an amplitude image and a phase image of the sample, and the computer 18 displays the amplitude image and the phase image of the sample.
In this embodiment, the preferred laser is a 532nm green laser with a power of 20mW and an attenuation factor of 0.01 for attenuator 2.
The focal length of the second lens 6 is larger than that of the first lens 4, the focal points of the two lenses are overlapped, and the focal length of the first lens 4 is 10-200 mm, preferably 50-150 mm; the focal length of the second lens 6 is 200 to 1000mm, preferably 300 to 800mm, and in this embodiment, the focal length of the first lens 4 is selected to be 100mm, and the focal length of the second lens is selected to be 500 mm.
The diaphragm 5 is arranged at the focus of the coincidence of the first lens 4 and the second lens 6, the diaphragm 5 is arranged on the three-dimensional displacement platform and can move up and down in the direction of the light beam generated by the laser, the size of the diaphragm is adjusted, the light beam does not pass through the diffraction, and stray light in a light path is blocked.
The first adjustable attenuation sheet 8 can adjust the attenuation multiple through rotation.
The microspheres 9 and the microsphere platform 19 are placed on a two-dimensional displacement platform which can move two-dimensionally in the plane perpendicular to the light beam, and can move two-dimensionally in the direction of the light beam generated by the laser, with micron precision, back and forth, left and right. The microsphere platform 19 is a transparent toughened film, and small holes with the diameter equal to that of the microspheres 9 are arranged on the microsphere platform. The microsphere 9 is barium titanate microsphere with diameter of 50 μm, the microsphere platform 19 is a 20 μm transparent toughened film, and the diameter of the small hole arranged thereon is 50 μm.
The sample 11 is placed on a sample stage, the sample stage is a three-dimensional moving platform and can move left and right, front and back and up and down in the direction of a light beam generated by a laser, the displacement precision is 50 nm-400 nm, preferably 50 nm-300 nm, and the displacement precision of the sample stage selected in the embodiment is 50 nm.
The embodiment discloses a single pixel scanning super-resolution phase imaging device, when using:
the first step is as follows: opening the laser, and moving the microspheres and the microsphere platform to the center of the laser beam;
the second step is that: adjusting the size of the small hole of the diaphragm, and moving the diaphragm to eliminate the stray light of the light path;
the third step: adjusting the first adjustable attenuation sheet to reduce high-order diffraction spots except the nano-photon jet flow;
the fourth step: moving the sample close to the microspheres;
the fifth step: recording diffraction images on a CCD image sensor, reconstructing amplitude and phase images of the sample, and displaying the amplitude and phase images of the sample by a computer;
specifically, an iterative algorithm is used to reconstruct an amplitude and phase image of the sample from the diffraction image.
And a sixth step: the sample is scanned in two dimensions with a displacement stage, and the amplitude and phase of the sample are reconstructed at each position and combined into an amplitude and phase image of the entire sample.
The single-pixel scanning super-resolution phase imaging device disclosed by the embodiment realizes the display of the amplitude image and the phase image of the sample.
Example 2
In the embodiment, a single-pixel scanning super-resolution phase imaging device is disclosed, a planar light beam is adopted to irradiate a microsphere array to generate a nano photon jet array; irradiating a sample by using a nano photon jet flow array to generate a diffraction signal, transmitting the signal to a CCD image sensor by using a lens, and recording an amplitude image of the sample; the spectroscope divides out another beam of plane laser, and the plane laser and the diffraction signal generate interference on the CCD image sensor to obtain a phase image of the sample; and (3) moving the sample by adopting a nanometer precision displacement platform, and scanning the sample to obtain a complete image of the sample.
The single-pixel scanning super-resolution phase imaging device disclosed in the embodiment has a structure shown in fig. 2, and includes a laser 1, an attenuation sheet 2, a polarizer 3, a first lens 4, a diaphragm 5, a second lens 6, a first spectroscope 7, a first adjustable attenuation sheet 8, a microsphere array 9, a microsphere array platform 10, a sample 11, a third lens 12, a second spectroscope 13, a CCD image sensor 17 and a computer 18, which are coaxially arranged in sequence along the advancing direction of a light beam; a first reflector 14, a second reflector 15 and a second adjustable attenuation sheet 16 are sequentially arranged along the advancing direction of the light beam split by the first spectroscope.
Light beams emitted by the laser 1 pass through the attenuation sheet 2, the polarizer 3, the first lens 4, the diaphragm 5, the second lens 6, the first spectroscope 7, the first adjustable attenuation sheet 8, the microsphere array 9, the microsphere array platform 10, the sample 11, the third lens 12 and the second spectroscope 13 to form diffraction signals, and the diffraction signals are recorded by the CCD image sensor 17 to obtain an amplitude image of the sample; the light beam split by the first spectroscope 7 enters the second spectroscope 13 after passing through the first reflective mirror 14, the second reflective mirror 15 and the second adjustable attenuation sheet 16, an interference signal is generated by the second spectroscope 13, the diffraction signal and the interference signal generate interference on the CCD image sensor 17, a phase image of the sample is obtained, and the amplitude image and the phase image of the sample are displayed by the computer 18.
In this embodiment, the preferred laser is a 532nm green laser with a power of 20mW and an attenuation factor of 0.01 for attenuator 2.
The focal length of the second lens 6 is larger than that of the first lens 4, the focal points of the two lenses are overlapped, and the focal length of the first lens 4 is 10-200 mm, preferably 50-150 mm; the focal length of the second lens 6 is 200 to 1000mm, preferably 300 to 800mm, and in this embodiment, the focal length of the first lens 4 is selected to be 100mm, and the focal length of the second lens is selected to be 500 mm.
The diaphragm 5 is arranged at the focus of the coincidence of the first lens 4 and the second lens 6, the diaphragm 5 is arranged on the three-dimensional displacement platform and can move up and down in the direction of the light beam generated by the laser, the size of the diaphragm is adjusted, the light beam does not pass through the diffraction, and stray light in a light path is blocked.
The first lens 4 and the second lens 6 are arranged in the advancing direction of a light beam generated by the laser, and when a sample is scanned to obtain an image, the light beam emitted by the laser is expanded, and then light spots cover all the transparent cut-off microspheres.
The microsphere array platform 10 is fixed on a two-dimensional displacement platform and can move two-dimensionally with micrometer precision in the direction of a light beam generated by a laser, and microspheres 9 form a microsphere array on the microsphere array platform 10.
The diameter of the microsphere 9 in the microsphere array platform 10 is 10 μm to 200 μm, preferably 20 μm to 50 μm, in this embodiment, the diameter of the microsphere is selected to be 50 μm, the microsphere may be selected from silica gel microsphere, glass microsphere and barium titanate microsphere, and the microsphere selected in this embodiment is barium titanate microsphere.
The microsphere array platform 10 is a transparent film, the single-layer microspheres are tightly arranged in the fence, and the thickness of the transparent film is smaller than the distance from the microsphere nano photon jet flow position to the microsphere surface.
The thickness of the microsphere array platform is smaller than the distance from the position of the nanometer photon jet flow to the surface of the microsphere.
The thickness of the microsphere array platform 10 is in the range of 100nm to 20 μm, preferably 10 μm to 20 μm, and in this embodiment the thickness of the microsphere array platform is selected to be 20 μm.
The first adjustable attenuation piece 8 and the second adjustable attenuation piece 16 are both rotary adjustable attenuation pieces, the light intensity is adjusted through the rotary attenuation pieces, in the process of scanning a sample, the sample moves close to the microsphere array platform, and the first adjustable attenuation piece and the second adjustable attenuation piece adopt the same attenuation times.
The first reflecting mirror 14 and the second reflecting mirror 15 are flat mirrors, and the first reflecting mirror 14 and the second reflecting mirror 15 are placed at an angle of 45 degrees with respect to the incident light beam.
In this embodiment, the number of pixels of the CCD image sensor is selected to be 656 × 492, and the pixel size is 5.6 μm.
The difference between this example and example 1 is: in this embodiment, on the basis of embodiment 1, a first beam splitter 7 is arranged between a second lens 6 and a first adjustable attenuation plate 8, and a light beam emitted by a laser is split into two beams by the first beam splitter 7, wherein one beam is used for obtaining an amplitude image and the other beam is used for obtaining a phase image; a first reflector 14, a second reflector 15 and a second adjustable attenuation sheet 16 for obtaining interference signals are added; a second beam splitter 13 is added to obtain parallel diffraction and interference signals.
This embodiment can obtain the amplitude image and the phase image of the sample directly by the CCD image sensor without performing calculation by the computer by modifying embodiment 1.
When the single-pixel scanning super-resolution phase imaging device disclosed by the embodiment is used:
selecting the thickness of the transparent film of the microsphere array platform according to the size and the refractive index of the microspheres;
selecting a first lens 4 and a second lens 6, wherein the focal length of the second lens 6 is greater than that of the first lens 4, the first lens 4 and the second lens 6 are placed on an optical bench, the focal points of the two lenses are overlapped, a diaphragm 5 is placed at the focal point of the two lenses, and the size of the diaphragm is adjusted to ensure that light beams do not pass through diffraction and stray light in a light path is blocked;
opening a laser, moving the microsphere array platform, enabling the light beam to uniformly irradiate on the microsphere array, adjusting the attenuation multiple of the attenuation sheet, and reducing the light intensity;
adjusting the attenuation multiple of the first adjustable attenuation sheet 8 to enable the light intensity of the position where the nano-photon jet flow is removed after the laser penetrates through the microsphere array to be approximately zero;
adjusting the attenuation times of the second adjustable attenuation sheet 16 to be the same as the attenuation times of the first adjustable attenuation sheet 8;
adjusting the spectroscope 7, the spectroscope 13, the reflective mirror 14 and the reflective mirror 15 to enable the two light beams to generate interference on the CCD image sensor;
using a nanometer precision displacement platform to move samples forwards and backwards and leftwards and rightwards, and recording amplitude images and phase images of the samples at different positions;
the amplitude and phase images at different positions are reconstructed to synthesize a total image of the sample.
Wherein, the focal position of the transparent medium microsphere can be calculated according to a spherical refraction formula; the thickness of the microsphere array platform film is smaller than the distance from the position of the nanometer photon jet flow to the surface of the microsphere, the operation is carried out under a microscope, the transparent medium microspheres are placed in the fence of the microsphere array platform, the single layer of the microspheres is tightly distributed, the microsphere array platform is horizontally fixed on the optical bench, and the transparent film is arranged below the microspheres.
The first lens 4 and the second lens 6 are arranged in the vertical direction, and after the laser is expanded, the light spot should cover all the transparent medium microspheres.
The specific process of adjusting the light beam to enable the light beam to be uniformly irradiated on the microsphere array comprises the following steps: and (3) closing the interference light, moving the lens 12 to enable the microsphere array platform to form a clear image on the CCD image sensor, moving the microsphere array platform by using the two-dimensional displacement platform, and observing the image recorded by the CCD image sensor until the microsphere array is uniformly illuminated.
The attenuation times of the first adjustable attenuation sheet 8 and the attenuation times of the attenuation sheets 2 should be adjusted at the same time, so that only the nanometer photon jet flow light spots irradiate on the sample.
The diffraction signal and the interference signal are incident on the CCD image sensor in parallel.
The moving step length of the displacement platform is less than or equal to the diameter of the nanometer photon jet flow light spot.
One amplitude image and one phase image are recorded at each scan position, and the amplitude and phase images at different positions are reconstructed to synthesize a total image of the sample.
In each sample position, firstly, under the condition of no interference light, an image of the CCD image sensor is recorded, an amplitude image of the sample at the position is obtained, then, under the condition of interference light, the image of the CCD image sensor is recorded, a phase image of the position can be calculated, the scanning frequency can be set according to requirements, and finally, the amplitude image and the phase image of each position of the sample are combined into a total image.
The single-pixel scanning super-resolution phase imaging device disclosed by the embodiment can obtain an amplitude image and a phase image of a sample, and can scan the sample to obtain a complete image of the sample by moving the sample stage, so that the technical problem that the conventional super-resolution imaging is only limited to intensity microscopy but cannot realize phase microscopy is solved.
Irradiating the microsphere array with light beams to generate a nano photon jet array; irradiating a sample by using a nano photon jet flow array to generate a diffraction signal, transmitting the signal to a CCD image sensor by using a lens, and recording an amplitude image of the sample; and the spectroscope divides out another beam of plane laser, the plane laser and the diffraction signal generate interference on the CCD image sensor to obtain a phase image of the sample, and the sample is moved by adopting a displacement platform to scan the sample to obtain a complete image of the sample.
The nano photon jet flow light spot of the transparent medium microsphere is used as a light source, the size of the light source is smaller than the limit resolution of a microscope, and the size and the light intensity of the light spot can be obtained according to a meter scattering theory.
The monolayer transparent medium microspheres are densely arranged, and the transparent film is arranged between the microsphere array and the sample, so that the irradiation of nanometer light spots on the sample is ensured, and the stable placement of the transparent medium microspheres is also ensured.
The transparent medium microspheres are used for super-resolution microscopy to obtain an absorption contrast image, a super-resolution phase image is not reported, under the condition of no interference light, the CCD image sensor records the absorption contrast image of a sample, namely an amplitude image of the sample, under the condition of interference light, the CCD image sensor records the phase contrast image of the sample, namely the phase image of the sample, the step length of a nano displacement platform is smaller than the size of a nano photon jet flow light spot, the super-resolution amplitude and the phase image of the sample are obtained through two-dimensional scanning, the resolution blank between an electron microscope and an optical microscope can be made up, and the transparent medium microspheres are important in scientific research, medical treatment and production life.
In the two light spots generated by the embodiment, the distance between the adjacent light spots is larger than the minimum resolution distance of the lens, the resolution is larger than that of the lens, the crosstalk between the two diffraction light spots is avoided, the size of the nano photon jet flow light spot can be determined by selecting the size and the refractive index of the transparent medium microsphere, and the resolution of the image is determined.
In the embodiment, transparent medium microspheres form a monolayer array which is tightly arranged, a planar light wave is used for irradiating the microsphere array, an imaging sample is tightly attached to the microsphere array, so that a nano photon jet flow array directly irradiates the sample, a CCD (charge coupled device) image sensor is used for recording an image, an amplitude image of the sample is obtained when no reference light exists, a phase image of the sample is obtained when the reference light exists, the sample is moved for scanning, and a complete amplitude image and a complete phase image of the sample are obtained. In the nano photon jet flow array, the distance of the nano photon jet flows is far greater than the minimum resolution distance given by Rayleigh criterion, so that diffraction spots of a sample at the nano photon jet flow position cannot be overlapped, the limitation of the resolution ratio of an optical device is broken, and super-resolution intensity and phase microscopy is realized. The research can make up the blank between the resolution of an electron microscope and the resolution of an optical microscope, and can be widely applied to scientific research, medical treatment and production life.
Example 3
In this embodiment, a single-pixel scanning super-resolution phase imaging method is disclosed, and a single-pixel scanning super-resolution phase imaging apparatus disclosed in embodiment 1 is used, including:
the laser emits a light beam;
the light beam forms a diffraction signal after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array and the sample, and the diffraction signal is recorded as a diffraction image by the CCD image sensor;
the computer obtains an amplitude image and a phase image of the sample from the diffraction image reconstruction by an iterative algorithm.
Example 4
In this embodiment, a single-pixel scanning super-resolution phase imaging method is disclosed, and a single-pixel scanning super-resolution phase imaging apparatus disclosed in embodiment 2 is used, including:
the laser emits a light beam;
the light beam passes through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first spectroscope, the first adjustable attenuation sheet, the microsphere array, the sample, the third lens and the second spectroscope to form a diffraction signal, and the diffraction signal is recorded by the CCD image sensor to obtain an amplitude image of the sample;
the light beam split by the first spectroscope enters the second spectroscope after passing through the first reflector, the second reflector and the second adjustable attenuation sheet, an interference signal is generated by the second spectroscope, and the diffraction signal and the interference signal generate interference on the CCD image sensor to obtain a phase image of the sample.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
As will be appreciated by one skilled in the art, embodiments of the present application may be provided as a method, system, or computer program product. Accordingly, the present application may take the form of an entirely hardware embodiment, an entirely software embodiment or an embodiment combining software and hardware aspects. Furthermore, the present application may take the form of a computer program product embodied on one or more computer-usable storage media (including, but not limited to, disk storage, CD-ROM, optical storage, and the like) having computer-usable program code embodied therein.
The present application is described with reference to flowchart illustrations and/or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of the application. It will be understood that each flow and/or block of the flow diagrams and/or block diagrams, and combinations of flows and/or blocks in the flow diagrams and/or block diagrams, can be implemented by computer program instructions. These computer program instructions may be provided to a processor of a general purpose computer, special purpose computer, embedded processor, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, create means for implementing the functions specified in the flowchart flow or flows and/or block diagram block or blocks.
These computer program instructions may also be stored in a computer-readable memory that can direct a computer or other programmable data processing apparatus to function in a particular manner, such that the instructions stored in the computer-readable memory produce an article of manufacture including instruction means which implement the function specified in the flowchart flow or flows and/or block diagram block or blocks.
These computer program instructions may also be loaded onto a computer or other programmable data processing apparatus to cause a series of operational steps to be performed on the computer or other programmable apparatus to produce a computer implemented process such that the instructions which execute on the computer or other programmable apparatus provide steps for implementing the functions specified in the flowchart flow or flows and/or block diagram block or blocks.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting the same, and although the present invention is described in detail with reference to the above embodiments, those of ordinary skill in the art should understand that: modifications and equivalents may be made to the embodiments of the invention without departing from the spirit and scope of the invention, which is to be covered by the claims.
Claims (10)
1. A single-pixel scanning super-resolution phase imaging device, comprising: the laser, the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array, the sample and the CCD image sensor are coaxially arranged in sequence along the advancing direction of the light beam, and the CCD image sensor is connected with the computer;
the light beam forms a diffraction signal after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array and the sample, the diffraction signal is recorded as a diffraction image by the CCD image sensor, and the amplitude image and the phase image of the sample are obtained by the reconstruction of the diffraction image by the computer.
2. The single-pixel scanning super-resolution phase imaging device according to claim 1, wherein the focal length of the second lens is larger than the focal length of the first lens, and the focal points of the first lens and the second lens coincide.
3. The single-pixel scanning super-resolution phase imaging device according to claim 2, wherein the diaphragm is placed at a focal point where the first lens and the second lens coincide.
4. The single-pixel scanning super-resolution phase imaging device according to claim 1, wherein the second adjustable attenuator and the first adjustable attenuator use the same attenuation factor.
5. A single-pixel scanning super-resolution phase imaging device, comprising: the laser, the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first spectroscope, the first adjustable attenuation sheet, the microsphere array, the sample, the third lens, the second spectroscope and the CCD image sensor are coaxially arranged in sequence along the advancing direction of a light beam, and the first reflector, the second reflector and the second adjustable attenuation sheet are sequentially arranged along the advancing direction of the light beam split by the first spectroscope;
light beams generated by the laser form diffraction signals after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first spectroscope, the first adjustable attenuation sheet, the microsphere array, the sample, the third lens and the second spectroscope, and the diffraction signals are recorded by the CCD image sensor to obtain an amplitude image of the sample; the light beam split by the first spectroscope enters the second spectroscope after passing through the first reflector, the second reflector and the second adjustable attenuation sheet, an interference signal is generated by the second spectroscope, and the diffraction signal and the interference signal generate interference on the CCD image sensor to obtain a phase image of the sample.
6. The single-pixel scanning super-resolution phase imaging device according to claim 5, wherein the interference signal and the diffraction signal are incident on the CCD image sensor in parallel.
7. The single-pixel scanning super-resolution phase imaging device according to claim 5, wherein the microspheres form a microsphere array on a microsphere array platform, the microsphere array platform is a transparent film, and the thickness of the microsphere array platform film is smaller than the distance from the position of the nano-photon jet stream to the surface of the microspheres.
8. The single-pixel scanning super-resolution phase imaging device according to claim 5, wherein the sample is placed on a sample stage, the sample stage is a three-dimensional displacement stage, and the sample is moved by the three-dimensional displacement stage.
9. A single-pixel scanning super-resolution phase imaging method is characterized by comprising the following steps:
the laser emits a light beam;
the light beam forms a diffraction signal after passing through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first adjustable attenuation sheet, the microsphere array and the sample, and the diffraction signal is recorded as a diffraction image by the CCD image sensor;
the computer obtains an amplitude image and a phase image of the sample from the diffraction image reconstruction.
10. A single-pixel scanning super-resolution phase imaging method is characterized by comprising the following steps:
the laser emits a light beam;
the light beam passes through the attenuation sheet, the polarizer, the first lens, the diaphragm, the second lens, the first spectroscope, the first adjustable attenuation sheet, the microsphere array, the sample, the third lens and the second spectroscope to form a diffraction signal, and the diffraction signal is recorded by the CCD image sensor to obtain an amplitude image of the sample;
the light beam split by the first spectroscope enters the second spectroscope after passing through the first reflector, the second reflector and the second adjustable attenuation sheet, an interference signal is generated by the second spectroscope, and the diffraction signal and the interference signal generate interference on the CCD image sensor to obtain a phase image of the sample.
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