CN111122510A - Transmission type orthogonal polarization phase microscopic imaging device based on F-P interferometer - Google Patents
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Abstract
The invention provides an intracavity enhancement type transmission orthogonal polarization phase microscopic imaging device based on an F-P interferometer. The method is characterized in that: the device consists of a laser light source 1, a light attenuation system 2, a laser beam expanding system 3, an F-P interferometer 4, a PBS (polarization beam splitter) prism 5, microscope objectives 6 and 8, detection cameras 7 and 9 and a computer 10. The invention can be used for digital holographic measurement of tiny objects and can be widely applied to the field of refractive index three-dimensional microscopic imaging in various objects.
Description
(I) technical field
The invention relates to an intracavity enhanced transmission type orthogonal polarization phase microscopic imaging device based on an F-P interferometer, which can be used for refractive index three-dimensional microscopic imaging in various tiny objects such as optical fibers, cells and the like, and belongs to the technical field of optical imaging.
(II) background of the invention
Microscopic Optical imaging, also commonly referred to as "Optical Microscopy" or "Light Microscopy", refers to a technique whereby a magnified image of a microscopic sample can be obtained after visible Light transmitted through or reflected from the sample has passed through one or more lenses. The resulting image can be viewed directly by eye through an eyepiece, recorded with a plate or digital image detector such as a Charge Coupled Device (CCD), Complementary Metal Oxide Semiconductor (CMOS), displayed on a computer, and analyzed.
There are three general limitations to conventional optical microscopy using bright field illumination: firstly, only imaging can be carried out on a dark color sample (transmission type) or a strong reflection type sample (reflection type); secondly, the maximum resolution of the technology is limited to about 200nm by the limit of optical diffraction; third, out-of-focus information can reduce image contrast. Fluorescence Microscopy (Fluorescence Microscopy) based on excitation and Fluorescence emission of fluorescent molecules in a sample (exogenous or endogenous) can break through the limitation that transparent samples cannot be imaged, but the problems of limited resolution and defocusing interference still need to be solved by adopting other measures.
In the 30 s of the 20 th century, the dutch physicist Zernike (Zernike) first proposed a phase contrast technique, whose principle is that the complex amplitude distribution on the image plane is approximately proportional to the phase distribution of the object by changing the phase of the direct light (i.e. zero frequency light) by ± 90 ° and attenuating it appropriately, so that the direct light and the diffracted light interfere, and the "invisible" phase change is converted into a "visible" intensity distribution. In a specific optical path, the zernike phase contrast microscope requires a ring-shaped condenser capable of producing cone-shaped illumination light, and a phase ring corresponding to the cone-shaped illumination light passing region at the back focal plane of the objective lens. With this technique, direct observation and imaging of a stain-free live cell sample can be conveniently achieved, but it has a disadvantage that it is not suitable for thick samples and very small samples.
Digital holography in recent years has been applied to microscopic imaging because it can record and display all information of recorded objects, wherein a three-wavelength reflection type digital holographic microscope consisting of three linearly polarized light sources differing in wavelength and a beam splitter prism, a lens, etc. is disclosed in patent CN 109062018A. Compared with the prior microscopic imaging device, the system only uses the reflected light as the interference light, cannot eliminate errors through contrast, and has a complex structure and is difficult to operate.
Patent CN109615651A discloses a three-dimensional microscopic imaging method and system based on a light field microscopic system, which performs three-dimensional reconstruction on a light field intensity image and the first forward projection matrix, the high resolution intensity image and the second forward projection matrix therein by using a preset algorithm to generate a three-dimensional reconstruction result of a three-dimensional sample. By adding one path of acquisition light path, the reconstruction signal-to-noise ratio of the focal plane reconstruction is enhanced under the same iteration times, the reconstruction effect of the light field microscopic imaging is greatly improved, but the method and the system are obtained by using more optimized algorithm reconstruction, and the structure of the relied light path is still unchanged.
Patent CN109520988A discloses a microscopic imaging system, vibration isolation platform, movable slide glass, imaging assembly. The system can be used for detecting different types of cell samples with high precision, but the system is used for imaging according to the principle of cell fluorescence, cannot be used for imaging other non-cell particles and objects, and has a small application range.
Patent CN201710904860.1 discloses an optical coherence tomography imaging system, which adopts mach-zehnder interference optical path, and is characterized in that optical fiber is adopted to simplify the system and reduce the cost, but the optical path structure is still more complex compared with the optical path structure of F-P cavity.
Patent CN201810145657.5 discloses a high resolution digital holographic diffraction tomography, which is characterized in that a mach-zehnder interference optical path structure is adopted, and a synthetic aperture method is used to obtain N synthetic high resolution holograms, thereby obtaining a high resolution three-dimensional refractive index representation of a measured sample. The structure is relatively more complex, and the invention is essentially different from the invention.
Patent cn201910136421.x discloses a super-resolution digital holographic imaging system and an imaging method, and the imaging system is characterized in that a transmission-type spatial light modulator is added in front of a traditional mach-zehnder interference optical path to modulate a light source. Compared with the invention adopting the optical path structure of the F-P cavity, the invention has essential difference.
In contrast, in the F-P interferometer intracavity enhancement type transmission orthogonal polarization phase microscopic imaging device designed by the invention, the fineness of the F-P interferometer is more than 20, so that multiple reflection of light beams in the F-P interferometer cavity can be utilized, and after the multiple reflection, coherent fringes generated by transmitted light are multiplied, compared with algorithm reconstruction, the invention is substantially improved from the structure of a detection light path, and the F-P interferometer intracavity enhancement type transmission orthogonal polarization phase microscopic imaging device is mainly characterized in that: (1) an accurate F-P common optical path structure; (2) an enhancement method of multiple round-trip phase accumulation in an F-P cavity; the substantial improvement of the two aspects can lead the invention to lay a solid foundation for directly improving the quality of microscopic imaging and the measurement resolution on the information source head to be measured.
In addition, the intracavity enhancement type transmission orthogonal polarization phase microscopic imaging device based on the F-P interferometer can improve interference resolution, increase interference fringe visibility and reduce errors by utilizing the recorded information compensation principle of the orthogonal polarization state and comparing two images obtained by two interference lights which are mutually vertical in the polarization state. The method can overcome information loss caused by single polarization state recording and improve the fidelity of digital holography.
Disclosure of the invention
The invention aims to provide an intracavity enhancement type transmission orthogonal polarization phase microscopic imaging device based on an F-P interferometer, which has a simple and compact structure and is easy to operate and adjust.
The object of the invention is achieved in that the method comprises the following steps:
the transmission-type orthogonal polarization enhancement type phase microscopic imaging device is composed of a laser light source 1, a light attenuation system 2, a laser beam expanding system 3, an F-P interferometer 4, a PBS (polarizing beam splitter) prism 5, microscopic objective lenses 6 and 8, detection cameras 7 and 9 and a computer 10.
In the device, an object to be detected is placed in an F-P interferometer 4, light emitted by a laser light source 1 is attenuated by a light attenuation system 2, then expanded by a laser beam expansion system 3, and then enters the F-P interferometer 4, corresponding transmitted light is transmitted to a PBS polarization beam splitter 5 when the light beam is reflected once in a cavity of the F-P interferometer, after multiple reflections, the PBS beam splitter generates two light beams with vertical propagation directions and mutually vertical polarization states, and then the two light beams are transmitted to micro objective lenses 6 and 8 respectively, and after signals are received by detection cameras 7 and 9 respectively, the obtained signals are transmitted to a computer 10 for processing. The computer 10 compares the two sets of signals, and eliminates the error of a single image to obtain an ideal image.
The invention also has the following technical characteristics:
the F-P interferometer in the used optical path system is used as a device for generating the multiplied optical path difference, and the optical path difference can be multiplied every time when the light passes through the particles to be detected through multiple reflections in the F-P cavity, so that the width of the interference fringes is obviously increased, and the aim of improving the resolution ratio is fulfilled.
In the F-P cavity, light beams are reflected and transmitted for multiple times in the F-P cavity, the phase is enhanced, and the complex amplitude of the light beams finally penetrating through the F-P cavity is as follows:
the complex amplitude of the transmitted light is the complex amplitude of the incident light in the F-P cavity, the surface reflectivity of the inner sides of the two parallel plane glass plates of the F-P cavity is shown, and delta is the phase distribution of the cells to be detected.
Then when F-P cavity multiple beams interfere, the phase distribution obtained by digital holography is:
where n is the index of refraction of the medium within the cavity, d is the thickness of the F-P cavity, and λ is the composite wavelength of the light source.
The accumulation of the refractive index of the light beam passing through each point inside the object to be measured along the propagation direction is the phase distribution obtained by the digital hologram, when the refractive index difference between the inside of the object to be measured and the environment medium around the object to be measured is small, the optical path difference is the accumulation of the refractive index along the path direction of the light beam, and then the relationship between the phase distribution and the refractive index distribution of the object to be measured is as follows:
wherein n (x, y, z) is the refractive index distribution inside the cell 2 to be measured, the z-axis is the direction of the light beam propagation, and n0Is the refractive index of the surrounding medium surrounding the test cell 2.
The phase distribution diagram obtained by the method and the phase distribution diagram generally obtained by M-Z interference are shown in FIG. 3, compared with the traditional M-Z method, an image with a more obvious change rate can be obtained, the sharpness of interference fringes is obviously enhanced, and a particle holographic image reconstructed by a computer has higher resolution than that of the traditional method and is more suitable for fine measurement.
Preferably, the free spectral range FSR of the F-P interferometer is larger than 100GHz, the fineness F is larger than 20, and the loss is less than or equal to 3 dB.
Preferably, the micro objective used in the device is a high resolution objective. It has excellent correction and extremely high numerical aperture, and therefore has excellent resolution, contrast, and image flatness in observation and photomicrography.
The invention is based on the principle of compensation of orthogonal polarization recording information, simultaneously records two holograms with mutually vertical polarization states, and compares the two holograms by a computer. The method can overcome the defect of information loss possibly caused by distortion and uneven light intensity distribution in the material, and can obviously improve the resolution of the reproduced image.
The device of the invention is convenient for adding various devices for effectively controlling the parallelism and improving the reflectivity of the F-P interferometer on the F-P interferometer at the later stage due to less optical elements, and can be modified in large space.
The invention has the advantages of simple structure, high resolution, high accuracy, good imaging performance and the like. The resolution of the reproduced image can be significantly improved.
(IV) description of the drawings
FIG. 1 is a reflection type orthogonal polarization enhanced phase microscopic imaging device based on an F-P interferometer. The method is characterized in that: the device consists of a laser light source 1, a light attenuation system 2, a laser beam expanding system 3, an F-P interferometer 4, a PBS (polarization beam splitter) prism 5, microscope objectives 6 and 8, detection cameras 7 and 9 and a computer 10.
FIG. 2 is an embodiment of a reflective orthogonal polarization enhanced phase microimaging device based on an F-P interferometer.
FIG. 3 is a three-dimensional structure diagram of an F-P cavity in a transmission-type enhanced phase microimaging measurement system based on an F-P interferometer, wherein 1-1 and 1-2 are F-P cavities formed by wedge-shaped mirrors, and 2 is a parallel mirror for ensuring parallelism and cavity distance of the F-P cavities.
FIG. 4 is an external device case in which an F-P cavity is placed, in which 1 is a fixing hole in which the cavity length and parallelism of the F-P cavity can be adjusted with screws, 2 is an area for placing a matching fluid, 3 is a micro-hole for placing a minute object such as an optical fiber, and 4 is a device case.
FIG. 5 is a phase intensity contrast plot for M-Z interferometric imaging and reflective cross-polarization enhanced phase microscopy imaging based on an F-P interferometer of the present invention
(V) detailed description of the preferred embodiments
The invention is further illustrated below with reference to specific examples. But should not be taken as limiting the scope of the invention.
Referring to fig. 1, fig. 1 is a schematic diagram of a reflective orthogonal polarization enhanced phase microscope imaging device based on an F-P interferometer according to the present invention. It can be seen from the figure that the reflective orthogonal polarization enhancement type phase microscopic imaging device based on the F-P interferometer comprises a laser light source 1, a light attenuation system 2, a laser beam expanding system 3, an F-P interferometer 4, a PBS (polarization beam splitter) prism 5, microscope objectives 6 and 8, detection cameras 7 and 9 and a computer 10. The positional relationship of the above-described original is as follows:
the system comprises an attenuation system 2, a beam expanding system 3, an F-P interferometer 4, a PBS (polarization beam splitter) prism 5, a microscope objective 6 and a detection camera 7 which are arranged in sequence along the optical axis direction of output light of the laser light source 1, wherein another light path for light splitting is arranged above the PBS prism, and the other light path for light splitting is a microscope objective 8 and a detection camera 9 which are arranged in sequence. The beam splitting surface and the light path strictly form an angle of 45 degrees, light passing through an F-P interferometer is proportionally divided into two beams of light with mutually vertical polarization directions, cavities at two ends of the F-P interferometer are strictly parallel, and the micro objective lenses 6 and 8 increase the numerical aperture value by using oil immersion objective lenses. The associated computer 10 is connected to the associated detection cameras 7 and 9.
Referring to fig. 2, the method for implementing three-dimensional phase imaging enhancement on a particle original to be detected according to the present invention includes the following steps:
in the device, an object to be detected 11 is placed in an F-P interferometer 4, the positions of elements in a light path are adjusted, so that coherent light output by a laser light source 1 passes through an attenuation system 2 and a laser beam expanding system 3 and is split at a PBS (polarization beam splitter) prism 5, the light intensity ratio of two beams of light at the time is strict at 1:1, the position of the F-P interferometer is adjusted at the time, so that the light beams are reflected for multiple times in an F-P cavity where the object to be detected is located, a microscope 6 is adjusted, and a detection camera 7 is moved slowly and is located at a light back focal plane passing through the microscope objective 6. After adjusting the microscope objective 8 so that the light of the upper beam path can be received by the microscope objective 8, the detection camera 9 is slowly moved to be located at the back focal plane of the microscope objective 8. After the two detection cameras receive the signals, the obtained signals are transmitted to the computer 10. The computer 10 obtains two different interferograms to make up the corresponding details and reduce errors.
Referring to fig. 5 again, fig. 5 is a phase intensity contrast diagram of M-Z interferometric imaging and reflective orthogonal polarization enhanced phase microscopic imaging based on the F-P interferometer of the present invention, it is obvious that the phase intensity diagram obtained by the present invention can obtain an image with a more obvious rate of change than the conventional M-Z method, which indicates that the sharpness of interference fringes is significantly enhanced, and a particle holographic image reconstructed by using a computer has a higher resolution than the conventional method, and is more suitable for fine measurement.
The invention relates to a reflection type orthogonal polarization enhancement type phase microscopic imaging device based on an F-P interferometer, which uses the F-P interferometer with a fixed cavity length to reflect and enhance coherent light carrying phase information of a sample to be detected for multiple times, and simultaneously uses light generated by a PBS polarization splitting prism to generate two interference patterns which are obtained by the same light source and have completely vertical polarization directions. In the current various phase microscopic imaging devices, compared with other devices, the structure of the invention is simpler, and a plurality of limiting conditions are not needed.
The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or the change made by the technical personnel in the technical field on the basis of the invention are all within the protection scope of the invention. The protection scope of the invention is subject to the claims.
Claims (5)
1. An intracavity enhancement type transmission type orthogonal polarization phase microscopic imaging device based on an F-P interferometer. The method is characterized in that: the system consists of a laser light source 1, a light attenuation system 2, a laser beam expanding system 3, an F-P interferometer 4, a polarization beam splitter Prism (PBS)5, microscope objectives 6 and 8, detection cameras 7 and 9 and a computer 10. In the device, a particle to be detected is placed in an F-P interferometer (Fabry-Perot interferometer), light emitted by a laser light source 1 is attenuated by a light attenuation system 2, then expanded by a laser beam expansion system 3 and enters an F-P interferometer 4, corresponding transmitted light is transmitted to a PBS polarization beam splitter 5 after the light beam is reflected once in a cavity of the F-P interferometer, after multiple reflections, the PBS beam splitter generates two beams of light with vertical propagation directions and mutually vertical polarization states, the two beams of light are transmitted to microscope objectives 6 and 8 respectively, and after signals are received by detection cameras 7 and 9 respectively, the obtained signals are transmitted to a computer 10 for processing. The computer 10 compares the two sets of signals, and eliminates the error of a single image to obtain an ideal image.
2. A microscopic imaging apparatus according to claim 1, characterized in that: the F-P interferometer in the used optical path system is used as a device for generating the multiplied optical path difference, and the optical path difference can be multiplied every time when the light passes through the particles to be detected through multiple reflections in the F-P cavity, so that the width of the interference fringes is obviously increased, and the aim of improving the resolution ratio is fulfilled.
3. The F-P interferometer of claim 2, wherein: the length of the cavity of the F-P interferometer is unchanged, an object to be measured is placed in the cavity, and interference fringes amplified in multiples are obtained through multiple reflections.
4. A microscopic imaging apparatus according to claim 1, characterized in that: the PBS polarization beam splitter prism divides the light beam into two beams of light with vertical polarization directions, two images can be respectively obtained, information loss caused by single polarization state recording is overcome, and image resolution is improved.
5. The microscopic imaging measurement apparatus of claim 1. The method is characterized in that: the microscope objective used is a high-resolution objective.
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|---|---|---|---|---|
| CN113507775A (en) * | 2021-06-07 | 2021-10-15 | 中国工程物理研究院激光聚变研究中心 | Multipurpose optical Thomson scattering spectrum measuring system suitable for large laser device |
| CN113507775B (en) * | 2021-06-07 | 2023-11-21 | 中国工程物理研究院激光聚变研究中心 | Multipurpose optical Thomson scattering spectrum measuring system suitable for large-scale laser device |
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| CN111122510B (en) | 2023-07-21 |
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