CN216284937U - Efficient dual-channel circular polarization fluorescence spectrum measurement system - Google Patents
Efficient dual-channel circular polarization fluorescence spectrum measurement system Download PDFInfo
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- CN216284937U CN216284937U CN202122448953.2U CN202122448953U CN216284937U CN 216284937 U CN216284937 U CN 216284937U CN 202122448953 U CN202122448953 U CN 202122448953U CN 216284937 U CN216284937 U CN 216284937U
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Abstract
The utility model relates to the field of optical detection of samples, in particular to an efficient dual-channel circular polarization fluorescence spectrum measuring system, wherein the white light source, the sample to be measured, the objective lens, the dichroic mirror, the movable reflector, the 1/4 wave plate, the 50:50 beam splitter, the Glan prism, the high-pass filter, the focusing mirror and the Y-shaped optical fiber input end are arranged in a line in sequence, when the phases are formed by microscopy, the second movable reflector moves into the system, when the circular polarized fluorescence spectrum is measured, the second movable reflector moves out of the system, continuous laser emitted by the excitation light source is depolarized into natural light through the depolarizer, the natural light is reflected by the dichroic mirror and focused to a measured sample through the objective lens, the measured sample is excited to radiate circular polarized fluorescence, the circular polarized fluorescence is converted into linear polarized fluorescence through the 1/4 wave plate, and the linear polarized fluorescence is incident to the fluorescence spectrum acquisition system through the circular polarized fluorescence acquisition module. The utility model can simultaneously collect the left-handed circularly polarized fluorescence and the right-handed circularly polarized fluorescence of the tested sample, thereby greatly improving the system efficiency.
Description
Technical Field
The utility model relates to the field of optical detection of samples, in particular to an efficient dual-channel circular polarization fluorescence spectrum measurement system.
Background
The circular polarization fluorescence spectrum measuring system is a process of collecting the intensity of circular polarization fluorescence radiated simultaneously after exciting a sample molecular characteristic spectrum section by using depolarized exciting light. Detection of the circularly polarized fluorescence spectrum may assist in the analysis of the luminescent properties of the material.
In the reported circular polarization fluorescence spectrum measurement system, the excitation light source can be selected from a xenon lamp, an LED light source and the like, when the xenon lamp or the LED light source is adopted as excitation light, the spectrum coverage of the excitation light source is wide, even if the sample to be measured is excited in a focusing manner, the focus of the excitation light cannot be small enough to satisfy the circular polarization fluorescence spectrum measurement of the specific position of the unevenly distributed sample, and the requirement of in-situ in multiple measurements cannot be realized, in order to solve the above problems, it is conceivable to use a laser light source as an excitation light source, and since laser monochromaticity is good, a smaller focal diameter can be achieved when focusing, so that when the focal point is at a particular location in the sample being measured, the circular polarization fluorescence spectrum measurement can be selectively carried out on the region, and the in-situ circular polarization fluorescence spectrum measurement in multiple measurements can be realized by matching with a microscopic imaging technology.
In the polarization characteristic analysis of the circular polarization fluorescence spectrum, a left-handed fluorescence spectrum and a right-handed fluorescence spectrum in sample fluorescence are collected in sequence, and the asymmetric coefficient of the spectrum is obtained through mathematical analysis.
From the above analysis, it can be found that in the existing results, the measurement of the circular polarization fluorescence spectrum with high efficiency still remains to be solved.
SUMMERY OF THE UTILITY MODEL
The utility model aims to provide an efficient circular polarization fluorescence spectrum measuring system, which utilizes microscopic imaging to realize microscopic imaging and focusing of a sample area before measurement, realizes circular polarization fluorescence measurement of a non-uniformly distributed sample at a specific position by the method, and provides a method capable of simultaneously collecting a levorotation fluorescence spectrum and a dextrorotation fluorescence spectrum of a measured sample, thereby improving the collecting speed.
The technical scheme adopted by the utility model for realizing the purpose is as follows:
an efficient dual-channel circular polarization fluorescence spectrometry system, comprising:
the system comprises a laser light source generating system, a dichroic mirror, an objective lens, a white light source, a second movable reflector, an 1/4 wave plate, a 50:50 beam splitter, a first circular polarization fluorescence acquisition module, a second circular polarization fluorescence acquisition module, a Y-shaped optical fiber, a spectrum acquisition system and a microscopic imaging system;
wherein: the white light source, the objective lens, the dichroic mirror, the second movable reflector, the 1/4 wave plate, the 50:50 beam splitter and the first circularly polarized fluorescence collecting module are sequentially arranged in a line;
the light source generating system is arranged on an incident light path of the dichroic mirror, and a space for placing a sample to be detected is arranged between the objective lens and the white light source;
a microscopic imaging system is arranged on a reflected light path of the second movable reflector;
a first circular polarization fluorescence collection module is arranged on a transmission light path of the 50:50 beam splitter, a second circular polarization fluorescence collection module is arranged on a reflection light path, and the transmission light path and the reflection light path of the 50:50 beam splitter are perpendicular to each other; the first circular polarization fluorescence acquisition module and the second circular polarization fluorescence acquisition module are connected with the spectrum acquisition system through Y-shaped optical fibers.
When microscopic imaging is carried out on the sample to be measured, the second movable reflector is located between the dichroic mirror and the 1/4 wave plate, and white light emitted by the white light source illuminates the sample to be measured, so that the sample to be measured sequentially passes through the objective lens, the dichroic mirror and the second movable reflector and then is imaged in the microscopic imaging system.
When the circular polarization fluorescence spectrum measurement is carried out on the measured sample, the movable reflecting mirror II is moved out of the first line arrangement, the natural excitation light beam emitted by the laser source generation system sequentially passes through the dichroic mirror and the objective lens to irradiate on the measured sample, so that the measured sample emits the circular polarization fluorescence light beam after being excited, and the circular polarization fluorescence light beam sequentially passes through the objective lens, the dichroic mirror and the 1/4 wave plate, is split into the circular polarization fluorescence collection module I and the circular polarization fluorescence collection module II through the 50:50 beam splitter, and is transmitted to the spectrum collection system through the Y-shaped optical fiber.
The laser light source generating system comprises a first continuous laser light source, a first reflecting mirror, a second reflecting mirror, a third reflecting mirror and a depolarizer which are sequentially arranged on a continuous laser beam light path emitted by the first continuous laser light source, so that the continuous laser beam emitted by the first continuous laser light source is depolarized into a natural excitation beam through the depolarizer and then enters the dichroic mirror.
When the first movable reflector is arranged between the first reflector and the second reflector, the continuous laser beam emitted by the second continuous laser source sequentially passes through the first movable reflector, the second reflector, the third reflector and the depolarizer to be depolarized into a natural excitation beam to be emitted into the dichroic mirror.
The first circular polarization fluorescence collection module and the second circular polarization fluorescence collection module respectively comprise a Glan prism, a high-pass filter and a focusing mirror, and circular polarization fluorescence beams split by the 50:50 beam splitter are sequentially transmitted into the Y-shaped optical fiber through the Glan prism, the high-pass filter and the focusing mirror.
The microscopic imaging system comprises a fourth reflecting mirror, a third focusing mirror, a CCD camera, a second data transmission line and a second computer, wherein the CCD camera is connected with the second computer through the second data transmission line, and the fourth reflecting mirror, the third focusing mirror and the CCD camera are sequentially arranged on a reflecting light path of the second movable reflecting mirror.
The spectrum acquisition system comprises a CCD spectrometer, a first data transmission line and a first computer which are sequentially connected, wherein the CCD spectrometer is respectively connected with a first circularly polarized fluorescence acquisition module and a second circularly polarized fluorescence acquisition module through Y-shaped optical fibers.
The Y-shaped optical fiber comprises a Y-shaped optical fiber input end head I, a Y-shaped optical fiber input optical fiber I, a Y-shaped optical fiber input end head II, a Y-shaped optical fiber input optical fiber II, a Y-shaped optical fiber input end fusion point, a Y-shaped optical fiber output optical fiber and a Y-shaped optical fiber output end head, wherein the first circular polarization fluorescence acquisition module is connected with the Y-shaped optical fiber input end fusion point through the Y-shaped optical fiber input end head I and the Y-shaped optical fiber input optical fiber I in sequence; and the second circular polarization fluorescence acquisition module is connected with a Y-shaped optical fiber input end fusion point through a second Y-shaped optical fiber input end and a second Y-shaped optical fiber input end in sequence, and the Y-shaped optical fiber input end fusion point is connected with the spectrum acquisition system through a Y-shaped optical fiber output optical fiber and a Y-shaped optical fiber output end in sequence.
The tested sample is one of a spin-coated solid sample, a diamond anvil high-pressure module and a sample with the temperature lower than the set room temperature and environment temperature.
The utility model has the following beneficial effects and advantages:
1. the utility model is provided with a first movable reflector and a second movable reflector, wherein the first movable reflector can realize the function of switching excitation wavelength, when the first movable reflector moves into the system, a second continuous laser source is used as an excitation light source, when the second movable reflector moves out of the system, the first continuous laser source is used as an excitation light source, the second movable reflector can realize the function of switching between fluorescence spectrum acquisition and microscopic imaging, when the second movable reflector moves into the system, a detected sample is imaged in a microscopic imaging system, the microscopic imaging and micro-area focusing of the sample can be carried out, when the second movable reflector moves out of the system, the detected sample is excited by the excitation light source, and the spectrum acquisition system acquires the levorotatory fluorescence spectrum and the dextrorotatory fluorescence spectrum of the detected sample.
2. The utility model is provided with the first Glan prism and the second Glan prism, the optical axes of which are mutually vertical, and the included angles of the 1/4 wave plate optical axis in the system and the first Glan prism and the second Glan prism are both 45 degrees, so that the system can simultaneously acquire the levorotatory fluorescence spectrum and the dextrorotatory fluorescence spectrum of the tested sample, the acquisition efficiency of the system is improved, and the defect that the efficiency of acquiring the levorotatory fluorescence spectrum and the dextrorotatory fluorescence spectrum is low in the prior art is overcome.
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2a is a schematic diagram showing the relationship between the optical axis of a Glan prism and the optical axis of 1/4 wave plates in the circularly polarized fluorescence collection module of the present invention;
FIG. 2b is a schematic diagram showing the relationship between the optical axes of two Glan prisms and the optical axis of 1/4 wave plate in the circularly polarized fluorescence collecting module of the present invention;
FIG. 3a is a schematic diagram of the arrangement of the input and output ends of the Y-shaped optical fiber according to the present invention;
FIG. 3b is a schematic diagram of the arrangement of the input and output ends of the Y-shaped optical fiber according to the present invention;
FIG. 3c is a schematic diagram of the arrangement of the input and output ends of the Y-shaped optical fiber according to the present invention;
FIG. 4 is a schematic view of a CCD spectrometer of the present invention for spectral collection;
FIG. 5 is a schematic diagram of a circular polarization fluorescence spectrum of a measured sample obtained by the present invention;
wherein, 1 is a continuous laser light source I, 2 is a continuous laser light source II, 3 is a reflector I, 4 is a movable reflector I, 5 is a reflector II, 6 is a reflector III, 7 is a depolarizer, 8 is a dichroic mirror, 9 is an objective lens, 10 is a sample to be measured, 11 is a white light source, 12 is a movable reflector II, 13 is an 1/4 wave plate, 14 is a 50:50 beam splitter, 15 is a Glan prism I, 16 is a high pass filter I, 17 is a focusing mirror I, 18 is a Y-shaped optical fiber input end head I, 19 is a Y-shaped optical fiber input optical fiber I, 20 is a Glan prism II, 21 is a high pass filter II, 22 is a focusing mirror II, 23 is a Y-shaped optical fiber input end head II, 24 is a Y-shaped optical fiber input optical fiber II, 25 is a Y-shaped optical fiber input end welding point, 26 is a Y-shaped optical fiber output optical fiber, 27 is a Y-shaped optical fiber output end head, 28 is a CCD spectrometer, 29 is a first data transmission line, 30 is a first computer, 31 is a fourth reflector, 32 is a third focusing mirror, 33 is a CCD camera, 34 is a second data transmission line, 35 is a second computer, 181 is a cross-sectional enlarged schematic view of a first Y-type optical fiber input terminal 18, 221 is a cross-sectional enlarged schematic view of a second Y-type optical fiber input terminal 22, 271 is a cross-sectional enlarged schematic view of a Y-type optical fiber output terminal, 281 is a concave mirror, 282 is a beam splitting grating, 283 is a double row CCD array, and 2831 is an enlarged schematic view of a CCD array.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples.
As shown in fig. 1, the present invention includes 1, a first continuous laser light source, 1, a second continuous laser light source, 3, a first reflector, 4, a first movable reflector, 5, a second reflector, 6, a third reflector, 7, a depolarizer, 8, a dichroic mirror, 9, an objective lens, 10, a sample to be measured, 11, a white light source, 12, a second movable reflector, 13, 1/4 wave plate, 14,50:50 beam splitter, a first circular polarized fluorescence collection module, a second circular polarized fluorescence collection module, a Y-type optical fiber, a spectrum collection system and a microscopic imaging system, wherein the white light source 11, the sample to be measured 10, the objective lens 9, the dichroic mirror 8, the second movable reflector 12, 1/4 wave plate 13, the first circular polarized fluorescence collection module 14 and the first circular polarized fluorescence collection module are arranged in a line, i.e. arranged in sequence along the optical path direction, and when the microscopic imaging is performed on the sample to be measured 10, the second movable reflector 12 is moved into the system, white light emitted by a white light source 11 illuminates a sample 10 to be detected, the sample 10 to be detected is imaged in a microscopic imaging system after passing through an objective lens 9, a dichroic mirror 8 and a movable reflector II 12, when the circular polarization fluorescence spectrum measurement is carried out on the sample to be detected, the movable reflector 12 is moved out of the system, the movable reflector I4 switches a continuous laser light source 1 and a continuous laser light source II 2 according to requirements, when the movable reflector I4 is moved into the system, the continuous laser light source II 2 is used as an excitation light source, when the movable reflector I4 is moved out of the system, the continuous laser light source I1 is used as an excitation light source, after the excitation light source is selected, continuous laser light beams emitted by the excitation light source are reflected by a reflector II 5 and a reflector III 6 and then enter a depolarizer 7 and are depolarized into natural excitation light beams with non-polarization characteristics by the depolarizer 7, and the wavelength of the natural excitation light beams is lower than the threshold wavelength of the dichroic mirror 8, when the natural excitation light beam enters the dichroic mirror 8, the dichroic mirror 8 reflects the natural excitation light beam to the measured sample 10, the measured sample 10 is excited by the natural excitation light beam to emit a circular polarized fluorescent light beam, the wavelength of the circular polarized fluorescent light beam is higher than the threshold wavelength of the dichroic mirror 8 and is transmitted through the dichroic mirror 8, the circular polarized fluorescent light beam transmitted through the dichroic mirror 8 is transmitted through the 1/4 wave plate 13 and is converted into a linear polarized fluorescent light beam, the linear polarized fluorescent light beam enters the 50:50 beam splitter 14, the 50:50 beam splitter splits the linear polarized fluorescent light beam into a transmission linear polarized fluorescent light beam and a reflection linear polarized fluorescent light beam with equal energy, and the transmission linear polarized fluorescent light beam and the reflection linear polarized fluorescent light beam respectively enter the first circular polarized fluorescent light collecting module and the second circular polarized fluorescent light collecting module, the first circular polarized fluorescence collection module and the second circular polarized fluorescence collection module respectively collect the left-handed circular polarized fluorescence and the right-handed circular polarized fluorescence in the fluorescence beam excited to emit radiation by the tested sample 10, the left-handed circular polarized fluorescence and the right-handed circular polarized fluorescence are input into the first Y-shaped optical fiber input end 18 and the second Y-shaped optical fiber input end 23, the left-handed circular polarized fluorescence and the right-handed circular polarized fluorescence are input into the spectrum collection system after being transmitted by the Y-shaped optical fiber, and the spectrum collection system performs spectrum collection and analysis on the left-handed circular polarized fluorescence and the right-handed circular polarized fluorescence, so that the circular polarized fluorescence spectrum information of the tested sample 10 is finally obtained. The sample (10) to be tested can be a spin-coated solid sample, can be a diamond anvil high-pressure module, and can be a vacuum low-temperature sample. Wherein, the high pressure refers to the pressure higher than the standard atmospheric pressure in the environment of the tested sample, and the environment is provided by the diamond anvil. Low temperature refers to an ambient temperature below room temperature, which is provided by the vacuum chamber in conjunction with the cooling rod.
As shown in fig. 2a to 2b, when a sample to be measured is excited by excitation light with 10 times of excitation light to radiate a circularly polarized fluorescent light beam, the circularly polarized fluorescent light beam is composed of a left circularly polarized light beam and a right circularly polarized light beam, and the left circularly polarized light beam and the right circularly polarized light beam have different light-out rotation directions, so that after the left circularly polarized light beam and the right circularly polarized light beam transmit through 1/4 wave plate 13, they are converted into 2 linearly polarized light beams with mutually perpendicular polarization directions, and by adjusting the optical axis directions of glan prism one 15 and glan prism two 20, the optical axis directions of glan prism one 15 and glan prism two 20 are mutually perpendicular and have the maximum transmittance, at this time, the linearly polarized light beams transmitting through glan prism one 15 and glan prism two 20 respectively correspond to the left circularly polarized light speed and the right circularly polarized light beam of the circularly polarized fluorescent light beam.
As shown in fig. 3a to 3c, the first Y-shaped optical fiber input end 18 and the second Y-shaped optical fiber input end 23 are formed by welding 7 optical fibers, the 7 optical fibers are arranged in a circumferential manner, the Y-shaped optical fiber output end 27 is formed by welding 14 optical fibers, the 14 optical fibers respectively correspond to the 7 optical fibers of the first Y-shaped optical fiber input end 18 and the 7 optical fibers of the second Y-shaped optical fiber input end 23, the 14 optical fibers are arranged in a linear vertical manner, wherein the 7 optical fibers corresponding to the first Y-shaped optical fiber input end 18 are arranged on the upper portions of the 14 optical fibers arranged in a linear manner, and the 7 optical fibers corresponding to the second Y-shaped optical fiber input end 23 are arranged on the lower portions of the 14 optical fibers arranged in a linear manner.
As shown in fig. 1, after the linearly polarized fluorescent light beam transmitted through the glan prism i 15 is transmitted through the high pass filter 18, the excitation light beam doped in the linearly polarized fluorescent light beam is filtered, the linearly polarized fluorescent light beam is focused by the focusing lens 17 onto the gold Y-shaped optical fiber input end i 18, the optical fiber arrangement in the Y-shaped optical fiber input end i 18 is shown in fig. 3, the focused linearly polarized fluorescent light beam can be efficiently coupled into the Y-shaped optical fiber input end i 18, and the linearly polarized fluorescent light beam transmitted through the glan prism ii 20 is incident into the Y-shaped optical fiber input end i 23 according to the same principle.
As shown in fig. 4, the Y-fiber output terminal 27 is formed by fusing 14 optical fibers arranged linearly, the fluorescent light beam received by the first Y-fiber input terminal 18 and the second Y-fiber input terminal 23 is transmitted to the Y-fiber output terminal 27 through the Y-fiber, the Y-fiber output terminal 27 is coupled to the CCD spectrometer 28, the CCD spectrometer 28 is formed by a concave mirror 281, a beam splitter 282 and a dual-row CCD array 283, the divergent fluorescent light beam emitted from the Y-fiber output terminal 27 is collimated by the concave mirror 281 and transmitted to the beam splitter 282, the beam splitter 282 splits the collimated fluorescent light beam in the wavelength direction, the split fluorescent light beam is incident to the dual-row CCD array 283, the dual-row CCD array 283 detects the intensity of the split fluorescent light beam, the light intensity is converted into an electrical signal through photoelectric conversion, the fluorescent light beams output by the 7 optical fibers located at the upper portion of the Y-fiber output terminal 27 are detected by the dual-row CCD array 283, the fluorescent light beams output by the 7 optical fibers positioned at the lower part in the Y-shaped optical fiber output tip 27 are detected by the lower row array in the double-row CCD array 283, so that the double-row CCD array 283 can simultaneously detect the left-handed circularly polarized fluorescence and the right-handed circularly polarized fluorescence in the circularly polarized fluorescent light beams excited to radiate by the sample 10 to be detected.
As shown in fig. 1, the CCD spectrometer 28 simultaneously performs spectrum collection on the left-handed circularly polarized fluorescence and the right-handed circularly polarized fluorescence of the detected sample 10, the double-row CCD array 283 of the CCD spectrometer 28 respectively converts the optical signals of the left-handed circularly polarized fluorescence and the right-handed circularly polarized fluorescence of the detected sample 10 into electrical signals, and the electrical signals are processed by the first computer 30 to obtain the spectrum of the left-handed circularly polarized fluorescence and the spectrum of the right-handed circularly polarized fluorescence of the detected sample 10.
The working principle of the utility model is as follows:
as shown in fig. 1, in the microscopic imaging process, the second movable mirror 12 is moved into the system, the white light source 11 illuminates the sample 10 to be measured, the sample 10 to be measured passes through the objective lens 9, the dichroic mirror 8 and the movable mirror 12 and then is imaged in the microscopic imaging system, the microscopic imaging system is composed of a fourth mirror 31, a third focusing mirror 32, a CCD camera 33, a second data transmission line 34 and a second computer, the sample 10 to be measured is placed on a translation stage with three degrees of freedom of movement X, Y and Z, the degree of freedom of XYZ is adjusted by adjusting an adjusting knob of the translation stage in the microscopic imaging process, so as to adjust the position of the sample 10 to be measured, the sample 10 to be measured can be a clear and complete image on the CCD camera 33, and the clear and complete image is displayed by the second computer 35.
As shown in fig. 1, during the collection of the circular polarized fluorescence spectrum, two times of the movable mirror is moved out of the system, the movable reflector I4 is switched between excitation light sources by moving in or out of the system, when the movable reflector I4 moves in the system, the continuous laser light source II 2 is used as the excitation light source of the system, when the movable reflector one 4 moves out of the system, the continuous laser source one 1 is used as an excitation light source of the system, the continuous laser emitted by the excitation light source is reflected by the first reflecting mirror 5 and the second reflecting mirror 6 and then transmits through the depolarizer 7, the depolarizer depolarizes the continuous laser into a natural beam without polarization characteristic, the natural light beam is reflected by the dichroic mirror 8 and focused on a measured sample 10 by the objective lens 9, and the measured sample 10 is excited by the natural light beam to radiate circularly polarized fluorescence, wherein the circularly polarized fluorescence consists of left-handed circularly polarized fluorescence and right-handed circularly polarized fluorescence.
As shown in fig. 2 a-2 b, in the polarization conversion process of the circularly polarized fluorescence, the circularly polarized fluorescence is collimated by the objective lens 9, then transmitted through the dichroic mirror 8, and then incident to the 1/4 wave plate 13, the 1/4 wave plate 13 converts the circularly polarized fluorescence into the linearly polarized fluorescence with mutually perpendicular polarization directions, the linearly polarized fluorescence with mutually perpendicular polarization directions is split into a transmitted beam and a reflected beam with equal energy by the 50:50 beam splitter 14, the transmitted beam and the reflected beam are respectively incident to the left-handed circularly polarized fluorescence collecting module and the right-handed circularly polarized fluorescence collecting module, the glan prism 15 and the glan prism 20 in the left-handed circularly polarized fluorescence collecting module and the right-handed circularly polarized fluorescence collecting module respectively transmit the linearly polarized fluorescence with mutually perpendicular polarization directions, the linearly polarized fluorescence transmitted through the glan prism 15 and the glan prism 20 is filtered by the high pass filter 16 and the high pass filter 21, the linear polarized fluorescence with the wavelength larger than the threshold wavelengths of the high-pass filter 16 and the high-pass filter 21 can transmit through the high-pass filter 16 and the high-pass filter 21, and the linear polarized fluorescence transmitted through the high-pass filter 16 and the high-pass filter 21 is focused into the Y-shaped optical fiber input end 18 and the Y-shaped optical fiber input end 23 through the first focusing lens 17 and the second focusing lens 22.
As shown in fig. 3a to 3c, during the fluorescence collection and propagation process, the Y-shaped optical fiber consists of 2 input optical fibers and 1 output optical fiber, the input optical fiber is formed by welding 7 optical fibers, the 7 optical fibers are in an arrangement mode that 6 optical fibers are arranged around the center 1 optical fiber in a circumferential mode, the output optical fiber is formed by welding 14 optical fibers, the 14 optical fibers are respectively from the 2 input optical fibers, and the 14 optical fibers are arranged in a linear perpendicularity manner, in the fluorescence collection process, the linearly polarized fluorescence is focused into the Y-shaped optical fiber input end 18 and the Y-shaped optical fiber input end 23 through the first focusing lens 17 and the second focusing lens 22, the linearly polarized fluorescence focused into the Y-shaped optical fiber input end 18 and the Y-shaped optical fiber input end 23 is transmitted through the Y-shaped optical fiber, then is output through the Y-shaped optical fiber output end 27, and the linearly polarized fluorescence output through the Y-shaped optical fiber output end 27 is coupled and injected into the CCD spectrometer 28.
As shown in fig. 4, during the spectrum collection and analysis, after the fluorescence output from the Y-fiber output end 27 is coupled into the CCD spectrometer 28, the fluorescence is an angular divergent light beam with the Y-shaped optical fiber output end 27 as the vertex, the angular divergent light beam is collimated into a parallel light beam by the concave mirror 281 and then transmitted to the light splitting grating 282, the light splitting grating 282 performs horizontal diffraction light splitting on the parallel light beam in the wavelength direction, the horizontally diffracted and split light beams are transmitted to the double-row CCD array 283, the double-row CCD array 283 converts the light intensity information of the diffracted and split light beams into electric signals, because the 14 optical fibers in the Y-shaped optical fiber output terminal 27 are linearly and vertically arranged, after a series of collimation and diffraction, the light beam incident to the double-row CCD array 283 is wavelength-split in the horizontal direction, still remaining vertically aligned with the vertical arrangement of the 14 fibers of the Y-fiber output stub 27.
As shown in fig. 1, during the signal collection and processing, the CCD spectrometer 28 converts the input fluorescence light into an electrical signal, and the electrical signal is transmitted to the first computer 30 through the first data transmission line 29, and the first computer 30 processes and visualizes the electrical signal.
As shown in FIG. 5, the bottom curve shows the linearly polarized fluorescence spectrum corresponding to the left-handed circularly polarized fluorescence beam of the tested sample 10 for the software, wherein the abscissa is the relative intensity of the linearly polarized fluorescence spectrum with the wavelength and the ordinate is the corresponding wavelength, i.e. ILThe middle curve is the difference between the relative light intensity of the left-hand circularly polarized fluorescence beam spectrum and the relative light intensity of the right-hand circularly polarized fluorescence beam spectrum of the tested sample 10, i.e. Δ ═ IL-IRThe top curve is the asymmetry coefficient of the fluorescence spectrum of the sample being measured, i.e.
And analyzing the asymmetric coefficient to obtain the circular polarization fluorescence spectrum characteristic of the detected sample 10.
Before the circular polarization fluorescence spectrum measurement of the measured sample is carried out by using the method, the system calibration needs to be carried out by using the standard sample, and when the method is used for measuring the measured sample, the circular polarization fluorescence spectrum measurement of the measured sample is carried out after the microscopic imaging and the area selection focusing of the measured sample are finished.
In this embodiment, the first continuous laser light source 1 is preferably continuous laser with a wavelength of 355nm, the second continuous laser light source 2 is preferably continuous laser with a wavelength of 405nm, the dichroic mirror 8 is preferably continuous laser with a critical wavelength of 410nm, the objective lens 9 is preferably focal length of 200nm, the sample to be detected is preferably a diamond anvil high-voltage module, the white light source 11 is preferably an LED white light source, and the high-pass filter 16 and the high-pass filter 21 are preferably continuous laser with a critical wavelength of 430 nm.
Claims (10)
1. An efficient dual-channel circular polarization fluorescence spectrum measurement system is characterized by comprising:
the device comprises a laser light source generating system, a dichroic mirror (8), an objective lens (9), a white light source (11), a movable reflector II (12), an 1/4 wave plate (13), a 50:50 beam splitter (14), a circular polarized fluorescence acquisition module I, a circular polarized fluorescence acquisition module II, a Y-shaped optical fiber, a spectrum acquisition system and a microscopic imaging system;
wherein: the white light source (11), the objective lens (9), the dichroic mirror (8), the movable reflector II (12), the 1/4 wave plate (13), the 50:50 beam splitter (14) and the circular polarization fluorescence collection module I are sequentially arranged in a line;
the light source generating system is arranged on an incident light path of the dichroic mirror (8), and a space for placing a sample to be detected (10) is arranged between the objective lens (9) and the white light source (11);
a microscopic imaging system is arranged on a reflected light path of the second movable reflector (12);
a first circular polarization fluorescence collection module is arranged on a transmission light path of the 50:50 beam splitter (14), a second circular polarization fluorescence collection module is arranged on a reflection light path, and the transmission light path and the reflection light path of the 50:50 beam splitter (14) are perpendicular to each other; the first circular polarization fluorescence acquisition module and the second circular polarization fluorescence acquisition module are connected with the spectrum acquisition system through Y-shaped optical fibers.
2. The high-efficiency dual-path circular polarization fluorescence spectrum measuring system according to claim 1, wherein when microscopic imaging is performed on the measured sample, the second movable reflecting mirror (12) is located between the dichroic mirror (8) and the 1/4 wave plate (13), and the white light emitted by the white light source (11) illuminates the measured sample (10), so that the measured sample sequentially passes through the objective lens (9), the dichroic mirror (8) and the second movable reflecting mirror (12) and is imaged in the microscopic imaging system.
3. The efficient dual-path circular polarization fluorescence spectrum measurement system of claim 1, wherein when a sample to be measured is subjected to circular polarization fluorescence spectrum measurement, the second movable reflector (12) is shifted out of a line arrangement, a natural excitation light beam emitted by the laser source generation system sequentially passes through the dichroic mirror (8) and the objective lens (9) to irradiate on the sample to be measured (10), so that the sample to be measured (10) is excited to emit a circular polarization fluorescence light beam, and the circular polarization fluorescence light beam sequentially passes through the objective lens (9), the dichroic mirror (8) and the 1/4 wave plate (13), is split into the first circular polarization fluorescence collection module and the second circular polarization fluorescence collection module through the 50:50 beam splitter (14), and is transmitted to the spectrum collection system through the Y-shaped optical fiber.
4. The efficient dual-path circular polarization fluorescence spectrum measurement system of claim 1, wherein the laser source generation system comprises a first continuous laser source (1), and a first reflector (3), a second reflector (5), a third reflector (6) and a depolarizer (7) which are sequentially arranged on a path of a continuous laser beam emitted by the first continuous laser source (1), so that the continuous laser beam emitted by the first continuous laser source (1) is depolarized into a natural excitation beam through the depolarizer (7) and then enters the dichroic mirror (8).
5. The efficient dual-channel circular polarization fluorescence spectrum measurement system according to claim 4, further comprising a second continuous laser source (2) and a first movable reflector (4), wherein when the first movable reflector (4) is arranged between the first reflector (3) and the second reflector (5), a continuous laser beam emitted by the second continuous laser source (2) is depolarized into a natural excitation beam through the first movable reflector (4), the second reflector (5), the third reflector (6) and the depolarizer (7) in sequence and then enters the dichroic mirror (8).
6. The efficient dual-channel circular polarization fluorescence spectrum measurement system of claim 1, wherein the first circular polarization fluorescence collection module and the second circular polarization fluorescence collection module both comprise a Glan prism, a high-pass filter and a focusing mirror, and circular polarization fluorescence beams split by the 50:50 beam splitter (14) sequentially pass through the Glan prism, the high-pass filter and the focusing mirror and are emitted into the Y-shaped optical fiber.
7. The efficient dual-channel circular polarization fluorescence spectrum measurement system of claim 1, wherein the microscopic imaging system comprises a fourth reflecting mirror (31), a third focusing mirror (32), a CCD camera (33), a second data transmission line (34) and a second computer (35), wherein the CCD camera (33) is connected with the second computer (35) through the second data transmission line (34), and the fourth reflecting mirror (31), the third focusing mirror (32) and the CCD camera (33) are sequentially arranged on the reflecting light path of the second movable reflecting mirror (12).
8. The efficient dual-channel circular polarization fluorescence spectrum measurement system of claim 1, wherein the spectrum collection system comprises a CCD spectrometer (28), a first data transmission line (29) and a first computer (30) which are connected in sequence, and the CCD spectrometer (28) is respectively connected with the first circular polarization fluorescence collection module and the second circular polarization fluorescence collection module through Y-shaped optical fibers.
9. The efficient dual-channel circular polarization fluorescence spectrum measurement system according to claim 1, wherein the Y-shaped optical fiber comprises a first Y-shaped optical fiber input end (18), a first Y-shaped optical fiber input optical fiber (19), a second Y-shaped optical fiber input end (23), a second Y-shaped optical fiber input optical fiber (24), a fusion point of the input end of the Y-shaped optical fiber (25), a second Y-shaped optical fiber output optical fiber (26) and a fusion point of the input end of the Y-shaped optical fiber (27), wherein the first circular polarization fluorescence collection module is connected with the fusion point of the input end of the Y-shaped optical fiber (25) sequentially through the first Y-shaped optical fiber input end (18) and the first Y-shaped optical fiber (19); the second circular polarization fluorescence acquisition module is connected with a Y-shaped optical fiber input end fusion point (25) sequentially through a second Y-shaped optical fiber input end (23) and a second Y-shaped optical fiber input end (24), and the Y-shaped optical fiber input end fusion point (25) is connected with the spectrum acquisition system sequentially through a Y-shaped optical fiber output optical fiber (26) and a Y-shaped optical fiber output end (27).
10. The efficient dual-path circular polarization fluorescence spectrometry system of claim 1, wherein the sample (10) to be tested is one of a spin-coated solid sample, a diamond anvil high-voltage module, and a sample with a temperature lower than a set room temperature and environment temperature.
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| CN115046933B (en) * | 2022-07-13 | 2024-04-26 | 首都师范大学 | Micro-area circular dichroism spectrum and circular polarized luminescence testing device |
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