CN114280020A - Super-resolution microscopic imaging device and method based on spatial light modulator - Google Patents
Super-resolution microscopic imaging device and method based on spatial light modulator Download PDFInfo
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Abstract
A super-resolution microscopic imaging device and method based on a spatial light modulator belong to the field of optical microscopic imaging. The device comprises a light source, a polarization beam splitter prism, an excitation light path, a loss light path and a coaxial system; a polarization beam splitter prism is arranged in front of the light source, an excitation light path is arranged at an outlet on one side of the polarization beam splitter prism, and a loss light path is arranged at an outlet on the other side of the polarization beam splitter prism; the outlet side of the excitation light path is coupled into a coaxial system; the loss optical path outlet is coupled into a coaxial system; one side of the coaxial system is provided with an imaging system, and the other side of the coaxial system is provided with a sample to be tested. Has the advantages that: the vortex light beam adjusted in real time can adjust the resolution of the whole system under the condition of not changing the existing light path structure, and the system can be quickly adjusted according to different application scenes; simple structure, ultrahigh resolution, dynamically adjustable resolution and the like.
Description
Technical Field
The invention relates to a super-resolution microscopic imaging device and method based on a spatial light modulator, and belongs to the field of optical microscopic imaging.
Background
The resolution of conventional optical microscopes is limited to about half a wavelength due to diffraction limitations. In the last two decades, many super-resolution imaging technologies that bypass the optical diffraction limit by different methods have appeared, in which stimulated radiation loss microscopy (STED) suppresses the luminescence of fluorescent molecules at the periphery of a fluorescent spot by introducing a beam of annular loss light, so as to achieve the purpose of reducing the point spread function and achieve super-resolution imaging. The invention of the super-resolution microscopy has very important significance in the fields of biomedicine, materials science and the like, however, the traditional STED system consists of an excitation light path and a loss light path, two beams of light are coupled into a coaxial microscopic scanning system together, the whole STED structure is huge, the resolution ratio of the system is inversely proportional to the integral microscopic scanning speed, an optical element in the loss light needs to be replaced in a scene needing different resolution ratios, and the integral light path needs to be adjusted to be coaxial after the element is replaced, so that the STED system is very inconvenient.
Disclosure of Invention
The invention aims to provide a super-resolution microscopic imaging device and method based on a spatial light modulator, which are simple in structure, convenient to operate, ultrahigh in resolution and dynamically adjustable.
In order to achieve the above object, the present application provides a super-resolution microscopic imaging device based on a spatial light modulator, which includes a light source, a polarization beam splitter prism, an excitation light path, a loss light path, and a coaxial system;
a polarization beam splitter prism is arranged in front of the light source, an excitation light path is arranged at an outlet on one side of the polarization beam splitter prism, and a loss light path is arranged at an outlet on the other side of the polarization beam splitter prism; the outlet side of the excitation light path is coupled into a coaxial system; the loss optical path outlet is coupled into a coaxial system; one side of the coaxial system is provided with an imaging system, and the other side of the coaxial system is provided with a sample to be tested.
Furthermore, the excitation light path is sequentially provided with a module, a polarizer, a beam splitter, a beam expanding and collimating system, a half-wave plate, 1/4 wave plate a, a beam splitter, a spatial light modulator, a third lens, a fourth lens and a dichroic mirror a.
Furthermore, the loss light path be equipped with filter a, first speculum, expand beam collimation module, first lens, second speculum, dichroic mirror b in proper order.
Furthermore, the coaxial system is sequentially provided with a focusing lens, a filter b, a dichroic mirror a, an 1/4 wave plate b and a microscope objective; the focusing lens is arranged on one side close to the imaging system.
Furthermore, the focusing lens, the filter b and the dichroic mirror b form an imaging light path; the dichroic mirror, the 1/4 wave plate and the microscope objective form a coaxial microscope scanning light path.
Further, the light source is a continuous spectrum laser.
Further, the third lens and the fourth lens form a 4f system, and the first lens and the second lens form a 4f system.
Furthermore, the imaging system is formed by splicing an imaging module and a subsequent data program.
The invention also discloses a super-resolution microscopic imaging method based on the spatial light modulator, which comprises the following steps:
s1, light beams generated by a light source are changed into excitation light and loss light through a polarization beam splitting prism.
S2, the exciting light is filtered, expanded and collimated, then passes through a 4f system consisting of a first lens and a second lens, and then is coupled into a coaxial system by a reflecting mirror and a dichroic mirror;
s3, the loss light enters a spatial light modulator after wavelength screening, filtering, beam expanding and collimation, the loss light is modulated by the spatial light modulator and then changed into vortex light beams, and the vortex loss light enters a coaxial system through a dichroic mirror coupling from a 4f system after passing through a 4f system consisting of a third lens and a fourth lens;
s4, focusing the excitation light and the loss light on a sample to be detected through a coaxial microscopic light path;
s5, the imaging light path is coaxial with the coaxial light path, then reaches the filter plate through the dichroic mirror, obtains tiny light spots only at the center of the vortex, and then is focused to the imaging system through the focusing lens.
Has the advantages that: compared with the traditional stimulated radiation loss microscopic system, the single light source is adopted to simplify the light path structure of the system, and meanwhile, the loss light is subjected to vortex modulation by the spatial light modulator, so that the size of the light vortex can be adjusted in real time; the vortex light beam adjusted in real time can adjust the resolution of the whole system under the condition of not changing the existing light path structure, and the system can be quickly adjusted according to different application scenes;
the super-resolution microscopic imaging device and method based on the spatial light modulator, which are provided by the invention, have great advantages in the fields of biomedicine and materials, and meanwhile, the method lays a foundation for simplification of coaxial optical system microscopic imaging and indicates the direction for real-time and rapid adjustment of super-resolution microscopic resolution;
the super-resolution microscopic imaging device and method based on the spatial light modulator have the advantages of being simple in structure, ultrahigh in resolution, dynamically adjustable in resolution and the like.
Drawings
Fig. 1 is a super-resolution microscopic imaging device based on a spatial light modulator.
The sequence numbers in the figures illustrate: 1. a light source; 2. a polarization beam splitter prism; 3. a module; 4. a polarizer; 5. a smooth wave; 6. a beam expanding collimation system; 7. a half-wave plate; 8. 1/4 wave plate a; 9. a beam splitter; 10. a spatial light modulator; 11. a third lens; 12. a fourth lens; 13. a dichroic mirror a; 14. 1/4 wave plate b; 15. a microscope objective; 16. a sample to be tested; 17. a filter segment a; 18. a first reflector; 19. a beam expanding and collimating module; 20. a first lens; 21. a second lens; 22. a second reflector; 23. a dichroic mirror b (23); 24. a filter b; 25. a focusing lens; 26. an imaging module.
Detailed Description
The invention will now be described in further detail with reference to the accompanying figure 1 and specific examples: the present application is further described by taking this as an example.
Example 1
As shown in fig. 1, the present embodiment provides a super-resolution microscopic imaging apparatus based on a spatial light modulator, which includes a light source 1, a polarization beam splitter prism 2, an excitation light path, a loss light path, and a coaxial system;
a polarization beam splitter prism 2 is arranged in front of the light source 1, an excitation light path is arranged at an outlet on one side of the polarization beam splitter prism 2, and a loss light path is arranged at an outlet on the other side of the polarization beam splitter prism 2; the outlet side of the excitation light path is coupled into a coaxial system; the loss optical path outlet is coupled into a coaxial system; one side of the coaxial system is provided with an imaging system, and the other side of the coaxial system is provided with a sample 16 to be measured.
The excitation light path is sequentially provided with a module 3, a polarizer 4, a diaphragm 5, a beam expanding and collimating system 6, a half-wave plate 7, a 1/4 wave plate a 8, a beam splitter 9, a spatial light modulator 10, a third lens 11, a fourth lens 12 and a dichroic mirror a 13.
The loss light path is sequentially provided with a filter a 17, a first reflector 18, a beam expanding and collimating module 19, a first lens 20, a second lens 21, a second reflector 22 and a dichroic mirror b 23.
The coaxial system is sequentially provided with a focusing lens 25, a filter plate b 24, a dichroic mirror b23, a dichroic mirror a 13, a 1/4 wave plate b 14 and a microscope objective lens 15; the focusing lens 25 is on the side near the imaging system.
The focusing lens 25, the filter b 24 and the dichroic mirror b23 form an imaging light path; the dichroic mirror 13, the 1/4 wave plate 14 and the microscope objective 15 form a coaxial microscope scanning light path.
The light source 1 is a continuous spectrum laser.
The third lens 11 and the fourth lens 12 form a 4f system, and the first lens 20 and the second lens 21 form a 4f system.
The imaging system is composed of an imaging module 26 and a subsequent data program.
A super-resolution microscopic imaging method based on a spatial light modulator comprises the following steps:
s1, light beams generated by a light source 1 are changed into excitation light and loss light through a polarization beam splitting prism.
S2, after filtering, beam expanding and collimating the exciting light, coupling the exciting light into a coaxial system through a reflecting mirror 22 and a dichroic mirror 23 after passing through a 4f system consisting of a first lens 20 and a second lens 21;
s3, the loss light enters the spatial light modulator 10 after wavelength screening, filtering, beam expanding and collimation, the loss light is modulated by the spatial light modulator 10 to become vortex light beams, and the vortex loss light is coupled into a coaxial system from a 4f system through a dichroic mirror 13 after passing through a 4f system consisting of a third lens 11 and a fourth lens 12;
s4, focusing the excitation light and the loss light on a sample 16 to be detected through a coaxial microscopic light path;
s5, the imaging light path is coaxial with the coaxial light path, then reaches the filter plate 24 through the dichroic mirror 23 to obtain a tiny light spot only at the center of the vortex, and then is focused to the imaging system through the focusing lens 25.
Example 2
In practical application, the light source 1 is a continuous spectrum laser; in the excitation light path, light beams emitted by the laser are split by the polarization beam splitter prism 2, pass through the filter 17 and the reflector 18, then enter the beam expanding and collimating module 19, pass through a 4f system consisting of the first lens 20 and the second lens 21, and then are coupled into a coaxial system by the reflector 22 and the dichroic mirror 23;
in the loss light path, the loss light is the other light split by the polarization beam splitter prism 2, the wavelength required by the loss light is screened out after passing through the wavelength selection module 3 and the polarizer 4, then the other light enters the polarization and phase modulation module consisting of the half-wave plate 7 and the 1/4 wave plate a 8 after passing through the optical wave 5 and the beam expanding collimation system 6, the modulated loss light enters the spatial light modulator 10, the vortex loss light modulated by the spatial light modulator enters the 4f system consisting of the third lens 11 and the fourth lens 12 through the beam splitter 9, and the vortex loss light is coupled into the coaxial system through the dichroic mirror 13 from the 4f system;
the coaxial scanning microscopic light path consists of an 1/4 wave plate b 14 and a microscope objective 15, and exciting light and loss light are focused on a sample 16 to be detected by the coaxial microscopic light path; the imaging light path consists of a dichroic mirror 23, a filter 24 and a focusing lens 25, the imaging light path is coaxial with the coaxial light path, then reaches the filter through the dichroic mirror to obtain tiny light spots only with vortex centers, and then is focused to an imaging system through the lens;
the imaging system is composed of an imaging module 26 and a subsequent data splicing program.
The foregoing is a more detailed description of the present invention in connection with specific preferred embodiments and is not intended to limit the practice of the invention to these embodiments. For those skilled in the art to which the invention pertains, several simple deductions or substitutions may be made without departing from the spirit of the invention, which should be construed as belonging to the protection of the present invention.
Claims (9)
1. A super-resolution microscopic imaging device based on a spatial light modulator is characterized by comprising a light source (1), a polarization beam splitter prism (2), an excitation light path, a loss light path and a coaxial system;
a polarization beam splitter prism (2) is arranged in front of the light source (1), an excitation light path is arranged at an outlet on one side of the polarization beam splitter prism (2), and a loss light path is arranged at an outlet on the other side of the polarization beam splitter prism (2); the outlet side of the excitation light path is coupled into a coaxial system; the loss optical path outlet is coupled into a coaxial system; one side of the coaxial system is provided with an imaging system, and the other side of the coaxial system is provided with a sample (16) to be measured.
2. The super-resolution microscopic imaging device based on the spatial light modulator according to claim 1, wherein the excitation light path is sequentially provided with a module (3), a polarizer (4), a diaphragm (5), a beam expanding and collimating system (6), a half-wave plate (7), a 1/4 wave plate a (8), a beam splitter (9), the spatial light modulator (10), a third lens (11), a fourth lens (12) and a dichroic mirror a (13).
3. The super-resolution microscopic imaging device based on the spatial light modulator according to claim 1, wherein the loss optical path is sequentially provided with a filter a (17), a first reflector (18), a beam expanding and collimating module (19), a first lens (20), a second lens (21), a second reflector (22), and a dichroic mirror b (23).
4. The super-resolution microscopic imaging device based on the spatial light modulator is characterized in that the coaxial system is sequentially provided with a focusing lens (25), a filter b (24), a dichroic mirror b (23), a dichroic mirror a (13), an 1/4 wave plate b (14) and a microscopic objective lens (15); the focusing lens (25) is arranged on one side close to the imaging system.
5. The spatial light modulator-based super-resolution microscopic imaging device according to claim 4, wherein the focusing lens (25), the filter b (24) and the dichroic mirror b (23) form an imaging optical path; the dichroic mirror (13), the 1/4 wave plate (14) and the microscope objective lens (15) form a coaxial microscope scanning light path.
6. The spatial light modulator-based super-resolution microscopy imaging device according to claim 1, wherein the light source (1) is a continuum laser.
7. The spatial light modulator-based super-resolution micro-imaging device according to claim 2, wherein the third lens (11) and the fourth lens (12) form a 4f system, and the first lens (20) and the second lens (21) form a 4f system.
8. The spatial light modulator-based super-resolution microscopy imaging device as claimed in claim 1, wherein the imaging system is composed of an imaging module (26) and a subsequent data program.
9. A super-resolution microscopic imaging method based on a spatial light modulator is characterized by comprising the following steps:
s1, light beams generated by the light source (1) are changed into excitation light and loss light through the polarization beam splitting prism.
S2, coupling exciting light into a coaxial system through a reflecting mirror (22) and a dichroic mirror (23) after the exciting light passes through a 4f system consisting of a first lens (20) and a second lens (21) after filtering, beam expanding and collimating;
s3, the loss light enters a spatial light modulator (10) after wavelength screening, filtering, beam expanding and collimation, the loss light is modulated by the spatial light modulator (10) to become vortex light beams, and the vortex loss light is coupled into a coaxial system from a 4f system through a dichroic mirror (13) after passing through a 4f system consisting of a third lens (11) and a fourth lens (12);
s4, focusing the excitation light and the loss light on a sample (16) to be detected through a coaxial microscopic light path;
s5, the imaging light path is coaxial with the coaxial light path, then reaches a filter plate (24) through a dichroic mirror (23), and obtains tiny light spots only with vortex centers, and then the tiny light spots are focused to an imaging system through a focusing lens (25).
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| CN116067935A (en) * | 2023-04-06 | 2023-05-05 | 北京攸维医疗科技有限公司 | Super-resolution imaging method and device for single-beam light path |
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