CN112903640A - Photon recoil imaging confocal detection system and method - Google Patents
Photon recoil imaging confocal detection system and method Download PDFInfo
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Abstract
The invention provides a photon recoil imaging confocal detection system and a photon recoil imaging confocal detection method, wherein the detection system emits an initial light source signal through an emission source and forms an adjusting light signal after passing through a wave selection amplification unit and a polarization adjusting unit, a beam splitter divides the adjusting light signal into a first light signal and a second light signal, the first light signal is focused on a sample to be detected by a first focusing unit, the sample to be detected generates an exciting light signal and a fluorescence signal, the exciting light signal and the fluorescence signal return to the beam splitter and coincide with the transmission direction of the second light signal, the second focusing unit focuses and filters the exciting light signal, the fluorescence signal and the second light signal, photons carrying sample information are separated from noise scattered photons by an atomic ultrasonic beam generator, and the atomic reaction mechanism of the sample to be detected is obtained by a preprocessing unit. The invention can distinguish photons carrying sample information from noise scattered photons with low momentum, obtain the reaction mechanism of the sample to be detected, effectively reduce the noise interference of a non-focal plane, and improve the spatial resolution and the imaging quality.
Description
Technical Field
The invention relates to the technical field of photon recoil imaging confocal, in particular to a photon recoil imaging confocal detection system and a photon recoil imaging confocal detection method.
Background
In the field of microscopic detection, increasing the detection thickness of a sample, improving the resolution and the imaging quality have been targets pursued by scientists. The confocal microscope forms a point light source by scanning light beams through a grating pinhole, scans point by point on a focal plane of a fluorescence-labeled specimen, collects optical signals of points to a receiver through a detection pinhole, and forms an image on a computer monitoring screen after signal processing. The light emitted from the focal point of the objective focal plane can be well converged at the pinhole, and can be completely received by the detector through the pinhole. Light emitted from the upper and lower positions of the focal plane can generate light spots with large diameters at the positions of the pinholes, and only a small part of light can penetrate through the pinholes and be received by the detector by comparing the diameters of the pinholes. And as the distance from the focal plane of the objective lens is larger, the stray light generated by the sample has larger scattered spot at the pinhole, and the energy capable of penetrating the pinhole is smaller (from 10% to 1%, slowly approaching 0%), so that the signal generated on the detector is smaller, and the influence is smaller. Just because confocal microscopy only images the focal plane of the sample, interference of diffracted light and scattered light is effectively avoided, so that the confocal microscope has higher resolution than a common microscope.
The conventional confocal microscope is optical signal imaging, and for the detection of a low-density diluted sample, a receiver is difficult to distinguish photons carrying important information from scattered noise photons, and the two photons are identical in momentum space. In the process of transmitting photon recoil imaging monitoring momentum from photons to atoms, stimulated Raman scattering is generated in an atom ultrasonic beam generator, and photons carrying detection information are directly detected, but a specific device for photon recoil imaging confocal detection does not exist in the prior art, and the photon recoil imaging confocal detection and the research of a target system mechanism and quantum nonlinear characteristics cannot be realized based on the specific device.
Disclosure of Invention
The invention aims to provide a photon back-flushing imaging confocal detection system and a photon back-flushing imaging confocal detection method, which can directly detect photons carrying sample information and the focusing depth of sample testing, obtain the reaction mechanism of a sample to be tested, distinguish photons carrying important information and noise scattered photons with low momentum, effectively reduce the interference of non-focal plane noise, and improve the spatial resolution and the imaging quality.
In order to achieve the purpose, the invention provides the following scheme:
a confocal detection system of photon back-flushing imaging is used for detecting a sample to be detected, and comprises:
an emission source for emitting an initial light source signal;
the wave selection amplifying unit is arranged on a transmission light path of the initial light source signal and is used for obtaining a wave selection amplifying light signal according to the initial light source signal;
the polarization adjusting unit is arranged on a transmission light path of the wave selection amplifying optical signal and is used for adjusting the wave selection amplifying optical signal and focusing the wave selection amplifying optical signal into a beam of light to obtain an adjusting optical signal;
the beam splitter is arranged on the transmission optical path of the adjusting optical signal and is used for splitting the adjusting optical signal into a first optical signal transmitted along the transmission optical path of the adjusting optical signal and a second optical signal vertical to the transmission direction of the first optical signal;
the first focusing unit is arranged on a transmission light path of the first optical signal and used for focusing the first optical signal on the sample to be detected; the first optical signal irradiates the sample to be detected, and the sample to be detected is excited by the first optical signal to generate an excitation optical signal and a fluorescence signal which carry sample information; the excitation light signal and the fluorescence signal return to the beam splitter in a direction opposite to a transmission optical path of the first light signal; the beam splitter is further used for deflecting the excitation light signals and the fluorescence signals returned to the beam splitter by 90 degrees so that the transmission directions of the excitation light signals and the fluorescence signals coincide with the transmission direction of the second light signals;
the second focusing unit is arranged on a transmission light path of the second optical signal and is used for obtaining a first filtered optical signal according to the exciting light signal at the beam splitter, obtaining a filtered fluorescent signal according to the fluorescent signal at the beam splitter and obtaining a second filtered optical signal according to the second optical signal;
the atomic ultrasonic beam generator is arranged on a transmission light path of the first filtered light signal, the filtered fluorescence signal and the second filtered light signal and is used for deflecting photons carrying the information of the sample to be detected in the first filtered light signal to obtain a first detection light signal, deflecting photons carrying the information of the sample to be detected in the filtered fluorescence signal to obtain a detection fluorescence signal and deflecting photons in the second filtered light signal to obtain a second detection light signal;
and the preprocessing unit is arranged on the transmission light path of the first detection light signal, the detection fluorescence signal and the second detection light signal and is used for obtaining the atomic reaction mechanism of the sample to be detected according to the first detection light signal, the detection fluorescence signal and the second detection light signal.
Optionally, the wave selection amplifying unit includes:
the first filter is positioned on a transmission light path of the initial light source signal and is used for filtering the initial light source signal to obtain a filtered light signal;
the half-wave plate is positioned on a transmission light path of the filtering light signal and used for polarizing the filtering light signal to obtain a polarized light signal;
the electro-optical modulator is positioned on a transmission light path of the polarized light signal and is used for carrying out pulse modulation on the polarized light signal to obtain a pulse light signal;
the first beam expander and the second beam expander are located on a transmission light path of the pulse light signals and used for collimating and expanding the pulse light signals to obtain wave-selecting amplified light signals.
Optionally, the polarization adjustment unit includes:
the spatial polaroid is positioned on a transmission light path of the wave selection amplified optical signal and is used for screening the wave selection amplified optical signal with set wavelength to obtain a screened optical signal;
the polarization analyzer is positioned on a transmission light path of the screening light signal and is used for monitoring and adjusting the screening light signal according to a set angle, intensity, phase and pulse width to obtain an adjusted light signal;
and the cylindrical mirror is positioned on the transmission light path of the adjusting light signal and is used for carrying out line focusing on the adjusting light signal to obtain an adjusting light signal.
Optionally, the first focusing unit includes:
the first lens is positioned on a transmission light path of the first optical signal and used for carrying out beam collimation and focusing on the first optical signal;
the first excitation galvanometer is positioned on a transmission light path of the first optical signal and used for deflecting the first optical signal by 90 degrees to obtain a deflected first optical signal;
the scanning mirror is positioned on a transmission light path of the deflected first optical signal and is used for performing rapid line scanning on the deflected first optical signal to obtain a scanned first optical signal;
the tube mirror is positioned on a transmission light path of the scanned first optical signal;
the objective lens is positioned on a transmission light path of the scanned first optical signal;
and the sample container is positioned on the transmission light path of the scanned first optical signal, positioned at the focal plane of the objective lens and used for containing the sample to be detected.
Optionally, the second focusing unit comprises:
the second lens is positioned on the transmission optical path of the second optical signal;
a third lens, located on the transmission optical path of the second optical signal, for enlarging the beam diameter in combination with the second lens;
the slit is positioned on a transmission light path of the second optical signal and is used for carrying out stray light filtration on the exciting light signal to obtain a first filtered optical signal, carrying out stray light filtration on the fluorescent signal to obtain the filtered fluorescent signal and carrying out stray light filtration on the second optical signal to obtain a second filtered optical signal;
a fourth lens, located on a transmission light path of the first filtered light signal, the filtered fluorescent signal, and the second filtered light signal, for collimating and focusing the first filtered light signal, the filtered fluorescent signal, and the second filtered light signal;
the second excitation galvanometer is positioned on a transmission light path of the first filtered light signal, the filtered fluorescent signal and the second filtered light signal and is used for deflecting the first filtered light signal by 90 degrees to obtain a deflected first filtered light signal, deflecting the filtered fluorescent signal by 90 degrees to obtain a deflected filtered fluorescent signal and deflecting the second filtered light signal by 90 degrees to obtain a deflected second filtered light signal;
and the fifth lens is positioned on a transmission light path of the deflected first filtered light signal, the deflected filtered fluorescent signal and the deflected second filtered light signal and is used for collimating and focusing the deflected first filtered light signal, the deflected filtered fluorescent signal and the deflected second filtered light signal.
Optionally, the second focusing unit further comprises:
and the second filter is positioned between the fourth lens and the second excitation galvanometer and is used for filtering the first filtered light signal, the filtered fluorescent signal and the second filtered light signal.
Optionally, the pre-processing unit comprises:
and the camera is used for imaging the first detection optical signal, the detection fluorescent signal and the second detection optical signal to obtain a photon image of the sample to be detected, and the photon image is used for representing an atomic reaction mechanism of the sample to be detected.
Optionally, the pre-processing unit comprises:
and the spectrometer is used for obtaining a composition spectrogram of the sample to be detected according to the first detection optical signal, the detection fluorescent signal and the second detection optical signal, and the composition spectrogram is used for representing an atomic reaction mechanism of the sample to be detected.
Optionally, the pre-processing unit comprises:
and the detector is used for obtaining a particle trajectory diagram of the sample to be detected according to the first detection optical signal, the detection fluorescent signal and the second detection optical signal, and the particle trajectory diagram is used for representing an atomic reaction mechanism of the sample to be detected.
In order to achieve the above purpose, the invention also provides the following scheme:
a photon recoil imaging confocal detection method is applied to the photon recoil imaging confocal detection system and comprises the following steps:
transmitting an initial light source signal through a transmission source;
receiving the initial light source signal through a wave selection amplifying unit, and obtaining a wave selection amplifying signal according to the initial light source signal;
adjusting the wave selection amplification signal through a polarization adjusting unit, and focusing the wave selection amplification signal into a beam of light to obtain an adjusting light signal;
dividing the adjusting optical signal into a first optical signal transmitted along a transmission optical path of the adjusting optical signal and a second optical signal vertical to the transmission direction of the first optical signal by a beam splitter;
focusing the first optical signal on the magnetic sample to be detected through a first focusing unit; the first optical signal irradiates the magnetic sample to be detected, and the magnetic sample to be detected is excited by the first optical signal to generate an excitation optical signal and a fluorescence signal which carry sample information; the excitation light signal and the fluorescence signal return to the beam splitter along the direction opposite to the transmission light path of the first light signal; the beam splitter is further used for deflecting the excitation light signals and the fluorescence signals returned to the beam splitter by 90 degrees so that the transmission directions of the excitation light signals and the fluorescence signals coincide with the transmission direction of the second light signals;
obtaining a first filtered light signal according to the excitation light signal deflected by 90 degrees by the beam splitter, obtaining a filtered fluorescence signal according to the fluorescence signal, and obtaining a second filtered light signal according to the second light signal by a second focusing unit;
deflecting photons carrying the information of the sample to be detected in the first filtered optical signal by an atomic ultrasonic beam generator to obtain a first detection optical signal, deflecting photons carrying the information of the sample to be detected in the filtered fluorescence signal to obtain a detection fluorescence signal, and deflecting photons in the second filtered optical signal to obtain a second detection optical signal;
and obtaining the atomic reaction mechanism of the sample to be detected according to the first detection optical signal, the detection fluorescent signal and the second detection optical signal through a preprocessing unit.
According to the specific embodiment provided by the invention, the invention discloses the following technical effects:
the invention provides a photon recoil imaging confocal detection system and a method, wherein an emission source emits an initial light source signal and forms an adjusting light signal after passing through a wave selection amplifying unit and a polarization adjusting unit, a beam splitter divides the adjusting light signal into a first light signal and a second light signal vertical to the transmission direction of the first light signal, a first focusing unit focuses the first light signal on a sample to be detected, the sample to be detected is excited by the first light signal to generate an exciting light signal and a fluorescence signal carrying sample information, the exciting light signal and the fluorescence signal return to the beam splitter along the direction opposite to the transmission light path of the first light signal, the beam splitter enables the transmission directions of the exciting light signal and the fluorescence signal to coincide with the transmission direction of the second light signal, a second focusing unit obtains a first filtering light signal according to the exciting light signal at the beam splitter, obtains a filtering fluorescence signal according to the fluorescence signal and obtains a second filtering light signal according to the second light signal, the atomic ultrasonic beam generator obtains a first detection light signal according to the first filtering light signal, obtains a detection fluorescence signal according to the filtering fluorescence signal and obtains a second detection light signal according to the second filtering light signal, and the preprocessing unit obtains an atomic reaction mechanism of the sample to be detected according to the first detection light signal, the detection fluorescence signal and the second detection light signal. The invention can directly detect the photons carrying the sample information and the focusing depth tested in the sample to obtain the reaction mechanism of the sample to be tested, can distinguish the photons carrying the sample information from noise scattered photons with low momentum, effectively reduces the interference of non-focal plane noise, and improves the spatial resolution and the imaging quality.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without inventive exercise.
FIG. 1 is a schematic diagram of a system structure of a confocal detection system for photon back-flushing imaging according to the present invention;
FIG. 2 is a schematic structural diagram of an atomic ultrasonic beam generator of the confocal detection system for photon back-flushing imaging according to the present invention;
FIG. 3 is a flow chart of the confocal detection method of photon recoil imaging according to the present invention.
Description of the symbols:
1-an emission source, 2-a wave selection amplifying unit, 21-a first filter, 22-a half-wave plate, 23-an electro-optical modulator, 24-a first beam expander and 25-a second beam expander;
3-polarization adjusting unit, 31-space polaroid, 32-polarization analyzer, 33-cylindrical mirror;
4-beam splitter, 5-first focusing unit, 51-first lens, 52-first excitation galvanometer, 53-scanning mirror, 54-tube mirror, 55-objective lens, 56-sample container;
6-second focusing unit, 61-second lens, 62-third lens, 63-slit, 64-fourth lens, 65-second filter, 66-second excitation galvanometer, 67-fifth lens;
7-atomic ultrasonic beam generator, 8-pretreatment unit.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention aims to provide a photon back-flushing imaging confocal detection system, which directly detects photons carrying sample information and the focusing depth tested in a sample, obtains the reaction mechanism of the sample to be tested, distinguishes the photons carrying the sample information from noise scattered photons, effectively reduces the interference of non-focal plane noise, and improves the spatial resolution and the imaging quality.
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in further detail below.
As shown in FIG. 1, the confocal detection system for photon recoil imaging of the present invention comprises: the device comprises an emission source 1, a wave selection amplifying unit 2, a polarization adjusting unit 3, a beam splitter 4, a first focusing unit 5, a second focusing unit 6, an atomic ultrasonic beam generator 7 and a preprocessing unit 8.
In particular, the emission source 1 is used to emit an initial light source signal.
The wave selection amplifying unit 2 is arranged on a transmission light path of the initial light source signal and is used for obtaining a wave selection amplifying light signal according to the initial light source signal.
The polarization adjusting unit 3 is arranged on a transmission light path of the wave selection amplifying optical signal and is used for adjusting the wave selection amplifying optical signal and focusing the wave selection amplifying optical signal into a beam of light to obtain an adjusting optical signal.
The beam splitter 4 is disposed on the transmission optical path of the conditioning optical signal, and is configured to split the conditioning optical signal into a first optical signal transmitted along the transmission optical path of the conditioning optical signal and a second optical signal perpendicular to the transmission direction of the first optical signal.
The first focusing unit 5 is disposed on a transmission light path of the first optical signal, and is configured to focus the first optical signal on a sample to be measured. The first optical signal irradiates a sample to be detected, and the sample to be detected is excited by the first optical signal to generate an excitation optical signal carrying sample information. The excitation light signal is returned to the beam splitter 4 in the opposite direction to the transmission optical path of the first light signal. The beam splitter 4 is also used to deflect the excitation light signal returning to the beam splitter 4 by 90 degrees such that the transmission direction of the excitation light signal coincides with the transmission direction of the second light signal.
The second focusing unit 6 is disposed on a transmission light path of the second optical signal, and is configured to obtain a first filtered optical signal according to the excitation light signal at the beam splitter 4, and obtain a second filtered optical signal according to the second optical signal.
The atomic ultrasonic beam generator 7 is arranged on the transmission light path of the first filtered light signal and the second filtered light signal, and is used for deflecting photons carrying information of a sample to be detected in the first filtered light signal to obtain a first detection light signal and deflecting photons in the second filtered light signal to obtain a second detection light signal.
The preprocessing unit 8 is disposed on a transmission light path of the first detection light signal and the second detection light signal, and is configured to obtain an atomic reaction mechanism of the sample to be detected according to the first detection light signal and the second detection light signal.
Specifically, the emission source 1 may be a laser transmitter with good coherence of light sources, such as a free electron laser, a higher harmonic generator, and an optical comb trigger.
Further, as shown in fig. 1, the wave selection amplifying unit 2 includes: a first filter 21, a half-wave plate 22, an electro-optical modulator 23, a first beam expander lens 24 and a second beam expander lens 25.
The first filter 21 is located on a transmission light path of the initial light source signal, and is configured to filter the initial light source signal to obtain a filtered light signal. The first filter 21 mainly filters light other than short wavelengths.
The half-wave plate 22 is located on a transmission light path of the filtered light signal, and is configured to polarize the filtered light signal to obtain a polarized light signal.
The electro-optical modulator 23 is located on a transmission light path of the polarized light signal, and is configured to perform pulse modulation on the polarized light signal to obtain a pulsed light signal. Specifically, since the polarized light signal is a continuous light signal, the electro-optical modulator modulates the continuous light signal into a pulsed light signal.
The first beam expander 24 and the second beam expander 25 are located on a transmission light path of the pulse light signals, and are used for collimating and expanding the pulse light signals to obtain wave-selecting amplified light signals.
Further, the polarization adjustment unit 3 includes: a spatial polarizer 31, a polarization analyzer 32, and a cylindrical mirror 33.
The spatial polarizer 31 is located on a transmission light path of the wavelength-selective amplified optical signal, and is configured to filter the wavelength-selective amplified optical signal to obtain a filtered optical signal. The spatial polarizer 31 allows only optical signals of a set wavelength to pass therethrough.
The polarization analyzer 32 is located on the transmission light path of the screening light signal, and is configured to monitor and adjust the screening light signal according to a set angle, intensity, phase, and pulse width to obtain an adjusted light signal. Wherein, can set up polarization angle, light intensity size, phase place and the pulse width of light signal according to actual need, combine relevant subassembly, polarization analyzer 32 can monitor and adjust the corresponding parameter of screening light signal according to above-mentioned parameter set value.
The cylindrical mirror 33 is located on the transmission optical path of the adjusting optical signal, and is used for performing line focusing on the adjusting optical signal to obtain an adjusting optical signal.
Further, the first focusing unit 5 includes: a first lens 51, a first excitation galvanometer 52, a scanning mirror 53, a tube mirror 54, an objective lens 55, and a sample container 56.
The first lens 51 is located on a transmission optical path of the first optical signal, and is configured to perform beam collimation and focusing on the first optical signal.
The first excitation galvanometer 52 is located on a transmission optical path of the first optical signal and is configured to deflect the first optical signal by 90 degrees to obtain a deflected first optical signal.
The scanning mirror 53 is located on the transmission light path of the deflected first optical signal, and is configured to perform fast line scanning on the deflected first optical signal to obtain a scanned first optical signal.
The tube mirror 54 is located on the transmission optical path of the scanned first optical signal.
The objective lens 55 is located on the transmission optical path of the scanned first optical signal.
The sample container 56 is located on the transmission light path of the scanned first optical signal, and located at the focal plane of the objective lens, and is used for containing a sample to be measured.
In order to obtain more accurate detection results, the second focusing unit 6 includes: a second lens 61, a third lens 62, a slit 63, a fourth lens 64, a second galvanometer 66, and a fifth lens 67.
The second lens 61 is located on a transmission optical path of the second optical signal.
The third lens 62 is located on the transmission path of the second optical signal, and is combined with the second lens for enlarging the beam diameter.
The slit 63 is located on a transmission light path of the second optical signal, and is configured to perform stray light filtering on the excitation optical signal and the second optical signal to obtain a first filtered optical signal and a second filtered optical signal. The size of the slit 63 determines the depth, resolution and imaging quality with which the sample to be measured can be detected.
The fourth lens 64 is located on the transmission light path of the first filtered light signal and the second filtered light signal, and is configured to collimate and focus the first filtered light signal and the second filtered light signal.
The second excitation galvanometer 66 is located on a transmission light path of the first filtered light signal and the second filtered light signal, and is configured to respectively perform 90-degree deflection on the first filtered light signal and the second filtered light signal to obtain a deflected first filtered light signal and a deflected second filtered light signal.
The fifth lens 67 is located on a transmission light path of the deflected first filtered light signal and the deflected second filtered light signal, and is configured to perform collimation and focusing on the deflected first filtered light signal and the deflected second filtered light signal.
To further optimize the above system, the second focusing unit 6 further comprises: a second filter 65.
The second filter 65 is disposed between the fourth lens and the second excitation galvanometer, and is configured to filter the first filtered optical signal and the second filtered optical signal.
Example 1:
in order to obtain image information of photon imaging of a sample to be measured, in embodiment 1 of the present invention, a preprocessing unit includes: a camera (not shown).
The camera is used for imaging the first detection optical signal and the second detection optical signal to obtain a photon image of the sample to be detected, and the photon image is used for representing an atomic reaction mechanism of the sample to be detected. Wherein the photons in the second detection light signal originate from the primary light source signal.
Example 2:
in order to obtain a spectrogram of an optical signal, in embodiment 2 of the present invention, the preprocessing unit includes: spectrometer (not shown).
The spectrometer is used for obtaining a composition spectrogram of the sample to be detected according to the first detection optical signal and the second detection optical signal, and the composition spectrogram is used for representing an atomic reaction mechanism of the sample to be detected. The second detection light signal contains photon information of the initial light source signal and can be used as reference light.
Example 3:
in order to obtain a travel locus diagram of a particle, in embodiment 3 of the present invention, the preprocessing unit includes: a detector (not shown).
The detector is used for obtaining a particle trajectory graph of the sample to be detected according to the first detection optical signal and the second detection optical signal, and the particle trajectory graph is used for representing an atomic reaction mechanism of the sample to be detected. Wherein the second detection light signal can be used as reference light.
On the basis of the embodiments 1, 2 and 3, the present invention provides a specific structural schematic diagram of an atomic ultrasonic beam generator in embodiment 4 for confocal detection of photon recoil imaging, as shown in fig. 2, specifically as follows:
example 4:
the principle is as follows: the two photons of the stimulated Raman scattering leave the atom along the same direction as the two incident photons, the momentum and the flight direction of the atom are not changed, and the stimulated Raman process can be clearly distinguished from other processes by detecting the flight direction of the atom.
Photons enter the atomic ultrasonic beam generator container 1903 from the atomic ultrasonic beam generator entrance window 1906, neon gas is sprayed from the atomic ultrasonic beam generator gas entrance port 1904, is collimated by the atomic ultrasonic beam generator atomic collimating element 1905 and interacts with the incident photons to generate stimulated raman scattering, the multichannel spatial detector 1907 detects the atomic spatial position, and the photons reach the grating spectrometer 1901 through the atomic ultrasonic beam generator exit window 1902.
As shown in fig. 3, to achieve the above object, the present invention further provides the following solutions:
a photon recoil imaging confocal detection method is applied to the photon recoil imaging confocal detection system and comprises the following steps:
an initial light source signal is emitted by the emission source.
And receiving the initial light source signal through a wave selection amplifying unit, and obtaining a wave selection amplifying signal according to the initial light source signal.
And adjusting the wave selection amplification signal through the polarization adjusting unit, and focusing the wave selection amplification signal into a beam of light to obtain an adjusted light signal.
The adjusting optical signal is divided into a first optical signal transmitted along a transmission optical path of the adjusting optical signal and a second optical signal perpendicular to the transmission direction of the first optical signal by a beam splitter.
The first optical signal is focused on the sample to be measured through the first focusing unit. The first optical signal irradiates a sample to be detected, and the sample to be detected is excited by the first optical signal to generate an excitation optical signal carrying sample information. The excitation light signal is returned to the beam splitter in a direction opposite to the transmission optical path of the first light signal. The beam splitter is also used for deflecting the excitation light signal returned to the beam splitter by 90 degrees so that the transmission direction of the excitation light signal coincides with the transmission direction of the second light signal.
And obtaining a first filtered light signal according to the excitation light signal deflected by 90 degrees by the beam splitter through the second focusing unit, and obtaining a second filtered light signal according to the second light signal.
And deflecting photons carrying information of the sample to be detected in the first filtered optical signal by an atomic ultrasonic beam generator to obtain a first detection optical signal, and deflecting photons in the second filtered optical signal to obtain a second detection optical signal.
And obtaining an atomic reaction mechanism of the sample to be detected according to the first detection optical signal and the second detection optical signal through a preprocessing unit.
The principle of the invention is as follows: the free electron laser emits short wavelength laser, and the short wavelength laser is input into the confocal micro-detection system after being amplified by wave selection. The method is characterized in that light with selected wavelength incident to a sample in a confocal microscopic system excites the sample, fluorescence generated by sample excitation returns to meet a slit through a light path, information such as stray light and diffraction light generated on a non-focal surface is blocked by the slit and cannot enter a receiver, only light with sample information carried on a focal plane enters a preprocessing unit, and part of noise scattering photons and photons with sample information simultaneously pass through the slit, so that the spatial resolution and the imaging quality are reduced.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other. The method disclosed by the embodiment corresponds to the system disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the system part for description.
The principles and embodiments of the present invention have been described herein using specific examples, which are provided only to help understand the method and the core concept of the present invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, the specific embodiments and the application range may be changed. In view of the above, the present disclosure should not be construed as limiting the invention.
Claims (10)
1. A confocal detection system of photon back-flushing imaging is used for detecting a sample to be detected, and is characterized by comprising:
an emission source for emitting an initial light source signal;
the wave selection amplifying unit is arranged on a transmission light path of the initial light source signal and is used for obtaining a wave selection amplifying light signal according to the initial light source signal;
the polarization adjusting unit is arranged on a transmission light path of the wave selection amplifying optical signal and is used for adjusting the wave selection amplifying optical signal and focusing the wave selection amplifying optical signal into a beam of light to obtain an adjusting optical signal;
the beam splitter is arranged on the transmission optical path of the adjusting optical signal and is used for splitting the adjusting optical signal into a first optical signal transmitted along the transmission optical path of the adjusting optical signal and a second optical signal vertical to the transmission direction of the first optical signal;
the first focusing unit is arranged on a transmission light path of the first optical signal and used for focusing the first optical signal on the sample to be detected; the first optical signal irradiates the sample to be detected, and the sample to be detected is excited by the first optical signal to generate an excitation optical signal and a fluorescence signal which carry sample information; the excitation light signal and the fluorescence signal return to the beam splitter in a direction opposite to a transmission optical path of the first light signal; the beam splitter is further used for deflecting the excitation light signals and the fluorescence signals returned to the beam splitter by 90 degrees so that the transmission directions of the excitation light signals and the fluorescence signals coincide with the transmission direction of the second light signals;
the second focusing unit is arranged on a transmission light path of the second optical signal and is used for obtaining a first filtered optical signal according to the exciting light signal at the beam splitter, obtaining a filtered fluorescent signal according to the fluorescent signal at the beam splitter and obtaining a second filtered optical signal according to the second optical signal;
the atomic ultrasonic beam generator is arranged on a transmission light path of the first filtered light signal, the filtered fluorescence signal and the second filtered light signal and is used for deflecting photons carrying the information of the sample to be detected in the first filtered light signal to obtain a first detection light signal, deflecting photons carrying the information of the sample to be detected in the filtered fluorescence signal to obtain a detection fluorescence signal and deflecting photons in the second filtered light signal to obtain a second detection light signal;
and the preprocessing unit is arranged on the transmission light path of the first detection light signal, the detection fluorescence signal and the second detection light signal and is used for obtaining the atomic reaction mechanism of the sample to be detected according to the first detection light signal, the detection fluorescence signal and the second detection light signal.
2. The confocal detection system of photon recoil imaging according to claim 1, wherein the wave selection amplification unit comprises:
the first filter is positioned on a transmission light path of the initial light source signal and is used for filtering the initial light source signal to obtain a filtered light signal;
the half-wave plate is positioned on a transmission light path of the filtering light signal and used for polarizing the filtering light signal to obtain a polarized light signal;
the electro-optical modulator is positioned on a transmission light path of the polarized light signal and is used for carrying out pulse modulation on the polarized light signal to obtain a pulse light signal;
the first beam expander and the second beam expander are located on a transmission light path of the pulse light signals and used for collimating and expanding the pulse light signals to obtain wave-selecting amplified light signals.
3. The confocal detection system of photon recoil imaging according to claim 1, wherein the polarization adjustment unit comprises:
the spatial polaroid is positioned on a transmission light path of the wave selection amplified optical signal and is used for screening the wave selection amplified optical signal with set wavelength to obtain a screened optical signal;
the polarization analyzer is positioned on a transmission light path of the screening light signal and is used for monitoring and adjusting the screening light signal according to a set angle, intensity, phase and pulse width to obtain an adjusted light signal;
and the cylindrical mirror is positioned on the transmission light path of the adjusting light signal and is used for carrying out line focusing on the adjusting light signal to obtain an adjusting light signal.
4. The confocal detection system of photon recoil imaging according to claim 1, wherein the first focusing unit comprises:
the first lens is positioned on a transmission light path of the first optical signal and used for carrying out beam collimation and focusing on the first optical signal;
the first excitation galvanometer is positioned on a transmission light path of the first optical signal and used for deflecting the first optical signal by 90 degrees to obtain a deflected first optical signal;
the scanning mirror is positioned on a transmission light path of the deflected first optical signal and is used for performing rapid line scanning on the deflected first optical signal to obtain a scanned first optical signal;
the tube mirror is positioned on a transmission light path of the scanned first optical signal;
the objective lens is positioned on a transmission light path of the scanned first optical signal;
and the sample container is positioned on the transmission light path of the scanned first optical signal, positioned at the focal plane of the objective lens and used for containing the sample to be detected.
5. The confocal detection system of photon recoil imaging according to claim 1, wherein the second focusing unit comprises:
the second lens is positioned on the transmission optical path of the second optical signal;
a third lens, located on the transmission optical path of the second optical signal, for enlarging the beam diameter in combination with the second lens;
the slit is positioned on a transmission light path of the second optical signal and is used for carrying out stray light filtration on the exciting light signal to obtain a first filtered optical signal, carrying out stray light filtration on the fluorescent signal to obtain the filtered fluorescent signal and carrying out stray light filtration on the second optical signal to obtain a second filtered optical signal;
a fourth lens, located on a transmission light path of the first filtered light signal, the filtered fluorescent signal and the second filtered light signal, for collimating and focusing the first filtered light signal, the filtered fluorescent signal and the second filtered light signal;
the second excitation galvanometer is positioned on a transmission light path of the first filtered light signal, the filtered fluorescent signal and the second filtered light signal and is used for deflecting the first filtered light signal by 90 degrees to obtain a deflected first filtered light signal, deflecting the filtered fluorescent signal by 90 degrees to obtain a deflected filtered fluorescent signal and deflecting the second filtered light signal by 90 degrees to obtain a deflected second filtered light signal;
and the fifth lens is positioned on a transmission light path of the deflected first filtered light signal, the deflected filtered fluorescent signal and the deflected second filtered light signal and is used for collimating and focusing the deflected first filtered light signal, the deflected filtered fluorescent signal and the deflected second filtered light signal.
6. The confocal detection system of photon recoil imaging according to claim 5, wherein the second focusing unit further comprises:
and the second filter is positioned between the fourth lens and the second excitation galvanometer and is used for filtering the first filtered light signal, the filtered fluorescent signal and the second filtered light signal.
7. The confocal detection system of photon recoil imaging according to claim 1, wherein the preprocessing unit comprises:
and the camera is used for imaging the first detection optical signal, the detection fluorescent signal and the second detection optical signal to obtain a photon image of the sample to be detected, and the photon image is used for representing an atomic reaction mechanism of the sample to be detected.
8. The confocal detection system of photon recoil imaging according to claim 1, wherein the preprocessing unit comprises:
and the spectrometer is used for obtaining a composition spectrogram of the sample to be detected according to the first detection optical signal, the detection fluorescent signal and the second detection optical signal, and the composition spectrogram is used for representing an atomic reaction mechanism of the sample to be detected.
9. The confocal detection system of photon recoil imaging according to claim 1, wherein the preprocessing unit comprises:
and the detector is used for obtaining a particle trajectory diagram of the sample to be detected according to the first detection optical signal and the second detection optical signal, and the particle trajectory diagram is used for representing an atomic reaction mechanism of the sample to be detected.
10. A confocal detection method of photon back-flushing imaging, which is applied to the confocal detection system of photon back-flushing imaging according to any one of claims 1 to 9, and comprises the following steps:
transmitting an initial light source signal through a transmission source;
receiving the initial light source signal through a wave selection amplifying unit, and obtaining a wave selection amplifying signal according to the initial light source signal;
adjusting the wave selection amplification signal through a polarization adjusting unit, and focusing the wave selection amplification signal into a beam of light to obtain an adjusting light signal;
dividing the adjusting optical signal into a first optical signal transmitted along a transmission optical path of the adjusting optical signal and a second optical signal vertical to the transmission direction of the first optical signal by a beam splitter;
focusing the first optical signal on the magnetic sample to be detected through a first focusing unit; the first optical signal irradiates the magnetic sample to be detected, and the magnetic sample to be detected is excited by the first optical signal to generate an excitation optical signal and a fluorescence signal which carry sample information; the excitation light signal and the fluorescence signal return to the beam splitter along the direction opposite to the transmission light path of the first light signal; the beam splitter is further used for deflecting the excitation light signals and the fluorescence signals returned to the beam splitter by 90 degrees so that the transmission directions of the excitation light signals and the fluorescence signals coincide with the transmission direction of the second light signals;
obtaining a first filtered light signal according to the excitation light signal deflected by 90 degrees by the beam splitter, obtaining a filtered fluorescence signal according to the fluorescence signal, and obtaining a second filtered light signal according to the second light signal by a second focusing unit;
deflecting photons carrying the information of the sample to be detected in the first filtered optical signal by an atomic ultrasonic beam generator to obtain a first detection optical signal, deflecting photons carrying the information of the sample to be detected in the filtered fluorescence signal to obtain a detection fluorescence signal, and deflecting photons in the second filtered optical signal to obtain a second detection optical signal;
and obtaining the atomic reaction mechanism of the sample to be detected according to the first detection optical signal, the detection fluorescent signal and the second detection optical signal through a preprocessing unit.
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