CN217738983U - Microscopic circular dichroism spectrum detection system based on single photon counting and collecting method - Google Patents
Microscopic circular dichroism spectrum detection system based on single photon counting and collecting method Download PDFInfo
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Abstract
The utility model belongs to the technical field of sample optical detection and specifically relates to a micro circular dichroism spectrum detection system based on single photon count collection method, including light source module, polarization module, arouse module, micro-imaging module, synchronization module, collection module and data processing module, wherein light source module produces the continuous light beam of broad spectrum, and polarization module will the continuous light beam polarization of broad spectrum is periodic variation's levogyration and right-handed polarized light, arouses in the module periodic variation's levogyration and right-handed polarized light take place the interaction, and collection module obtains light intensity signal after the interaction, data processing module will light intensity signal turns into corresponding circular spectral information through mathematical computation, in whole circular dichroism spectrum collection process, provides the time sequence information of control and collection by synchronization module, before carrying out circular dichroism spectrum collection, micro-imaging module can carry out micro-imaging to the sample of being surveyed, carries out the selection of interaction position simultaneously.
Description
Technical Field
The utility model belongs to the technical field of sample optical detection and specifically relates to a micro circular dichroism spectrum detection system based on single photon count collection method.
Background
Circular dichroism spectrum is an optical spectrum for deducing asymmetric molecular configuration and conformation, and the absorption coefficients of optically active substances for the left-handed and right-handed polarized light constituting plane polarized light are unequal, i.e. epsilon L ≠ε R The spectrum of the plane-polarized light is plotted as abscissa, and the difference in the absorptance (Δ = ∈) is plotted as abscissa L ≠ε R ) The spectrum obtained by taking the ordinate is the circular dichroism spectrum of the optically active substance, and the difference of the absorption rates can be obtained by obtaining the intensities of the left-handed polarized light and the right-handed polarized light which are transmitted through the optically active substance to be measured and performing mathematical calculation, so that the circular dichroism spectrum measuring system has the main task of measuring the intensities of the left-handed polarized light and the right-handed polarized light which are transmitted through the optically active substance to be measured.
In the results reported at present, the measurement of the light intensity of the left-handed and right-handed polarized light transmitted through the measured active substance is completed by adopting a modulation method, the method superposes periodic modulation signals with the same period as the change period of the left-handed and right-handed polarized light on the light intensity signals collected by a photodiode, and the method distinguishes the left-handed and right-handed polarized light components of the light intensity signals collected by a photomultiplier in one period, so that in the process of obtaining the left-handed and right-handed polarized light transmitted through the measured active substance, the signals must be modulated and demodulated, and the process has higher requirements on the sensitivity of related instruments, namely time accuracy, and can superpose system noise on the signals.
SUMMERY OF THE UTILITY MODEL
In order to solve the problem, the utility model discloses innovative adoption single photon counting method realizes the detection to the levogyration that transmits the photometry active material and the dextrorotation polarized light, adopts this kind of method can directly turn into digital signal with the light intensity to directly carry out digital computation to digital signal and can obtain corresponding circular dichroism spectrum.
In order to satisfy the circular dichroism spectrum's of the inhomogeneous sample of micro-zone sample and surface distribution survey, the utility model discloses set up micro-imaging module simultaneously, made the system can satisfy micro-zone and select district to measure.
An object of the utility model is to provide a micro circular dichroism spectral detection system based on single photon count collection system utilizes micro-imaging to realize the micro-imaging and focusing to the sample before the measurement, can realize the detection to the micro-district or the district selection circular dichroism of the sample of being surveyed.
The utility model discloses a realize that the technical scheme that above-mentioned purpose adopted is:
a micro circular dichroism spectrum detection system based on a single photon counting and collecting method comprises a light source module, a polarizing module, an excitation module and a collecting module which are sequentially arranged in a line, and further comprises a data processing module, a micro imaging module, a synchronization module and a white light source;
the data processing module is respectively connected with the acquisition module and the synchronization module through data transmission lines, the synchronization module is connected with the polarizing module through the data transmission lines, the microscopic imaging module is arranged on a refraction light path of the excitation module, and the white light source is arranged on a reflection light path of the excitation module.
The light source module comprises a broad spectrum continuous light source, and a first reflecting mirror and a second reflecting mirror which are sequentially arranged on a light path of the broad spectrum continuous light source.
The polarizing module comprises a polarizing film and an optical elastic modulator which are sequentially arranged in a line, the polarizing film and the reflector are arranged in a line, and the synchronous module is connected with the optical elastic modulator through a data transmission line.
The excitation module comprises: the device comprises a movable beam splitting sheet, a first reflective objective lens, a second reflective objective lens and a movable reflector which are sequentially arranged in a line, wherein the movable beam splitting sheet and the photoelastic modulator are arranged in a line, the microscopic imaging module is arranged on a refraction light path of the movable beam splitting sheet, the white light source is arranged on a reflection light path of the movable reflector, and a sample area for placing a sample to be measured is arranged between the first reflective objective lens and the second reflective objective lens.
The collecting module comprises a second focusing mirror and a grating light splitter which are sequentially arranged in a line, and further comprises a photomultiplier, the second focusing mirror and the movable reflecting mirror are arranged in a line, the photomultiplier is arranged on a reflection light path of the grating light splitter, and the photomultiplier is connected with the data processing module through a data transmission line.
The data processing module comprises a single photon counting and collecting card and a data processing computer, and the single photon counting and collecting card is respectively connected with the synchronization module, the data processing computer and the photomultiplier through a data transmission line.
The microscopic imaging module comprises a second focusing lens and a CCD camera which are sequentially arranged on a refraction light path of the movable beam splitting sheet, and further comprises an imaging display connected with the CCD camera through a data connecting line.
The synchronous module comprises a synchronous controller, and the synchronous controller is respectively connected with the photoelastic modulator and the single photon counting acquisition card through data connecting lines.
When the spectrum measurement is carried out on the tested sample, the movable beam splitting sheet and the movable reflector in the excitation system are arranged in a line, and the white light emitted by the white light source sequentially passes through the movable reflector and the reflective objective lens II in the excitation module to illuminate the sample area of the tested sample, so that the sample area of the tested sample sequentially passes through the reflective objective lens I and the movable beam splitting sheet and then is imaged in the microscopic imaging module.
When the spectrum of the tested sample is collected, the movable beam splitting sheet and the movable reflector in the excitation system are removed from one line, the wide-spectrum light beam emitted by the wide-spectrum continuous light source in the light source module sequentially passes through the polarizing module and the reflective objective lens in the excitation module to transmit the wide-spectrum light beam through the sample area of the tested sample, and the transmitted light sequentially passes through the reflective objective lens II and the collection module to be subjected to spectrum collection of the tested sample in the data processing module.
The utility model has the following beneficial effects and advantages:
1. the utility model discloses the novelty adopts single photon counting method to realize detecting the circular dichroism spectrum of the sample of being surveyed, has solved the tradition and has adopted the modulation method to carry out the circular dichroism spectrum and survey the relatively poor problem of time measuring signal to noise ratio, has promoted the system acquisition precision.
2. The utility model discloses be equipped with micro-imaging module, make the system have the function that micro-imaging was focused, expanded traditional system and can only carry out the technological blank that circular dichroism spectrum was gathered to the even sample of spatial distribution.
Drawings
FIG. 1 is a schematic view of the system of the present invention;
FIG. 2 is a schematic view of the present invention;
FIG. 3 is a schematic view of the working principle of the polarizing module of the present invention;
FIG. 4 is a schematic diagram of the timing control of the present invention;
the device comprises a wide-spectrum continuous light source 1, a first reflector 2, a second reflector 3, a polarizing plate 4, a photoelastic modulator 5, a movable beam splitting plate 6, a first reflective objective 7, a sample to be measured 8, a second reflective objective 9, a movable reflector 10, a first focusing mirror 11, a grating beam splitter 12, a photomultiplier 13, a white light source 14, a second focusing mirror 15, a CCD (charge coupled device) camera 16, a synchronous controller 17, a single-photon counting acquisition card 18, a data processing computer 19 and an imaging display 20.
501 is a photoelastic modulator controller.
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 utility model discloses a light source module, polarizing module, arouse module, micro-imaging module, synchronous module, collection module and data processing module, because the difference that the module constitutes causes the difference that measurement system constitutes, it is right here to combine the drawing the utility model discloses one of them system configuration does further detail.
As shown in fig. 2, the utility model comprises a light source module, a polarizing module, an excitation module, a microscopic imaging module, a synchronous module, an acquisition module and a data processing module, wherein the light source module comprises 1, a broad spectrum continuous light source, 2, a first reflector and a second reflector, the polarizing module comprises 4, a polarizer and 5, a photoelastic modulator, the excitation module comprises 6, a movable beam splitter, 7, a first reflective objective, 8, a sample to be measured, 9, a second reflective objective, 10, a movable reflector, the microscopic imaging module comprises 15, a second focusing mirror, 16, a CCD camera and 20, an imaging display, the synchronous module comprises a synchronous controller 17, the acquisition module comprises 11, a second focusing mirror, 12, a grating light splitter and 13, a photomultiplier, the data processing module comprises 18, a single photon counting acquisition card and 19, a data processing computer, in order to satisfy experimental requirements, the system simultaneously comprises 14 a white light source, wherein in the system, a second reflecting mirror 3, a polarizing plate 4, a photoelastic modulator 5, a movable beam splitting plate 6, a first reflecting objective 7, a sample to be measured 8, a reflecting objective 9, a movable reflecting mirror 10, a second focusing mirror 11 and a grating light splitter 12 are arranged in a line, the working process of the system comprises a microscopic imaging process and a spectrum acquisition process, when the movable beam splitting plate 6 and the movable reflecting mirror 10 are connected into the system, a light source module, a polarizing module, an excitation module, a microscopic imaging module and the white light source 14 participate in the microscopic imaging process, white light emitted by the white light source 14 sequentially passes through the movable reflecting mirror 10 and the second reflecting objective 9 to illuminate a sample area of the sample to be measured 8, and the sample area of the sample to be measured 8 sequentially passes through the first reflecting objective 7 and the movable beam splitting plate 6 to be imaged in the microscopic imaging module, when the movable beam splitter 6 and the movable reflector 10 are moved out of the system, a light source module, a polarization module, an excitation module, a synchronization module, an acquisition module and a data processing module participate in the spectrum acquisition process, a broad spectrum light beam emitted by a broad spectrum continuous light source 1 in the light source module enters the polarization module after changing the propagation direction through a reflector I2 and a reflector II 3, the broad spectrum light beam is polarized into periodically-changing left-handed and right-handed polarized light through a polarizer 4 and a photoelastic modulator 5 in the polarization module and then enters the excitation module, at the moment, the polarization module provides a trigger signal to the synchronization controller 17, the synchronization controller 17 respectively provides time delay for the acquisition module and the data processing module according to the specific conditions of the system to control the time sequence of the acquisition module and the data processing module, because the movable beam splitting sheet 6 and the movable reflector 10 in the excitation module are moved out of the system, the periodically-changed left-handed polarized light and the periodically-changed right-handed polarized light are focused by the reflective objective lens 7 and transmitted through a sample area of a sample 8 to be detected, the transmitted light is collimated into parallel light beams by the reflective objective lens II 9 and enters the acquisition module, the parallel light beams are focused to the grating beam splitter 12 by the focusing mirror II 11, the focusing light beams are split in the wavelength direction by the grating beam splitter and are diffracted to the photomultiplier tube 13, the photomultiplier tube 13 acquires the split single-wavelength light beams, the wavelength entering the photomultiplier tube 13 can be changed by changing the angle of the grating beam splitter 12, and the excitation light generated by the light source module is polarized into the periodically-changed left-handed polarized light and the periodically-changed right-handed polarized light by the polarization module, so that the photomultiplier tube 13 acquires the periodically-changed left-handed and right-handed polarized light, the photomultiplier 13 converts the collected optical signals into electrical signals to be transmitted to the data processing module, a single photon counting acquisition card 18 in the data processing module is responsible for controlling the time sequence in the acquisition process, so that the acquisition module can completely and accurately acquire the left-handed and right-handed polarized optical signals in one period, therefore, the left-handed and right-handed polarized optical signals with different wavelengths can be obtained after the diffraction wavelength is changed by changing the light splitting grating device 12, and the electrical signals converted by the acquisition module are calculated and arranged into circular dichroism spectra by the data processing computer 19.
The microscopic imaging process and the spectrum acquisition process are switched by a movable beam splitting sheet and a movable reflector.
The movable beam splitting piece and the movable reflector have two degrees of freedom of movement and rotation.
The light source module can be composed of a broad-spectrum continuous light source, a monochromator and a light reflecting element, and can also be composed of a broad-spectrum continuous light source and a light reflecting element.
The polarizing module is composed of a linear polarizing polarizer and a circular polarizing polarizer.
The linear polarization polarizer can be a linear polaroid or a Glan prism, and the circular polarization polarizer can be a rotatable 1/4 wave plate or an optical elastic modulator.
The excitation module is composed of a movable beam splitting sheet, a first reflective objective lens, a tested sample, a second reflective objective lens and a movable reflector, wherein the first reflective objective lens and the second reflective objective lens can be replaced by a transmission type focusing system.
The microscopic imaging module consists of a focusing lens, a camera and an imaging display, and in order to realize the microscopic imaging function, the system is also provided with a white light source.
The synchronization module is composed of a synchronization controller.
The collection module can comprise focusing mirror, grating beam splitter and photomultiplier, also can comprise focusing mirror and photomultiplier, when the light source module comprises wide spectrum continuous light source, monochromator and light reflection original paper, the collection module comprises focusing mirror and photomultiplier, when the light source module comprises wide spectrum continuous light source and light reflection original paper, the collection module comprises focusing mirror, grating beam splitter and photomultiplier.
The data processing module is composed of a single photon counting acquisition card 18 and a data processing computer 19.
As shown in fig. 3, the excitation light beam emitted from the broad-spectrum continuous light source 1 is natural light, that is, the polarization of the direction is the same, when the natural light enters the polarization module, the polarizer in the polarization module polarizes the natural light into linearly polarized light and then enters the photoelastic modulator 5, and the photoelastic modulator 5 exhibits the characteristic of a 1/4 wave plate with a periodically rotating optical axis under the control of the photoelastic modulator controller 501, so that the linearly polarized light becomes periodically changing left-handed and right-handed polarized light after passing through the photoelastic modulator 5.
As shown in fig. 4, in the spectrum collection process, the photoelastic modulator controller 501 generates a square wave signal with a frequency of 50kHz as a trigger signal of the photoelastic modulator 5, when the linearly polarized light transmitted through the polarizer 4 enters the photoelastic modulator 5, the photoelastic modulator 5 is represented as a 1/4 wave plate, the optical axis of the photoelastic modulator 5 periodically rotates under the control of the square wave signal, wherein the optical axis forms an angle of 45 ° with the polarization direction of the linearly polarized light, when the square wave signal is at the rising edge, the optical axis is located in two-four quadrants, and when the square wave signal is at the falling edge, the optical axis is located in one-three quadrants.
As shown in fig. 4, in the data processing process, a square wave signal with a frequency of 50kHz generated by the photoelastic modulator controller 501 triggers the synchronous controller 17, the synchronous controller 17 provides timing control signals to the acquisition module and the data processing module according to specific conditions of the system, and in order to distinguish left-handed and right-handed polarized light components corresponding to the acquired light intensity signals, the measured sample 8 has different absorptances to the left-handed and right-handed polarized light, so that the photomultiplier 13 acquires different light intensities in the optical axis rotation process of the photoelastic modulator 5, and further obtains electric signals with different intensities, the left-handed and right-handed polarized light components corresponding to the electric signals can be positioned by synchronizing the timing signals provided by the synchronous controller, and after the positions of the electric signals corresponding to the left-handed and right-handed polarized light are determined, the light intensity information is obtained by intercepting the integral signals within the same gating time.
The utility model discloses a theory of operation does:
as shown in fig. 2, the excitation module of the present invention is provided with a movable beam splitting plate 6 and a movable reflector 10, when microscopic imaging is performed on a sample 8 to be measured, the movable beam splitting plate 6 moves to a position between the photoelastic modulator 5 and the first reflective objective lens 7, the movable reflector 10 moves to a position between the second reflective objective lens 9 and the first focusing lens 11, in the microscopic imaging operation process, an excitation beam emitted from the broad spectrum continuous light source 1 in the light source module sequentially passes through the first reflective mirror 2, the second reflective mirror 3, the polarizer 4, the photoelastic modulator 5, the movable beam splitting plate 6 and the first reflective objective lens 7 and then focuses on a sample area of the sample 8 to be measured, a focus of the excitation beam coincides with the sample area, white light emitted from the white light source 14 sequentially passes through the movable reflector 10 and the second reflective objective lens 9 and then illuminates the sample area of the sample 8 to be measured, the sample area and the focus of the exciting light beam sequentially pass through a first reflective objective lens 7 and a movable beam splitting sheet 6 at the same time and then are reflected to enter a microscopic imaging module, a measured sample 8 is fixed on a translation stage with XYZ direction movement freedom, the position of the measured sample 8 is changed by adjusting the translation stage, so that the sample area of the measured sample 8 and the focus of the exciting light beam form a clear and complete image in the microscopic imaging module, an imaging display can display the clear and complete image, the superposition position of the focus of the exciting light beam and the sample area is changed by adjusting a first reflecting mirror 2 and a second reflecting mirror 3, and the excited position of the sample area is changed by the operation, so that the selected area of the measured sample 8 can be detected in the following spectral measurement process.
As shown in fig. 2, the utility model discloses an excitation module is equipped with portable beam splitter 6 and portable speculum 10, when carrying out spectral measurement to surveyed sample 8, portable beam splitter 6 and portable speculum 10 among the excitation module are shifted out the system, through the formation of image and the focus of microimaging process, the excitation position in surveyed sample 8 sample district has been selected, and at this moment, the light beam of the continuous light source 1 outgoing of broad spectrum reflects to polaroid 4 through speculum one 2 and speculum two 3, transmits polaroid 4 and photoelastic modulator 5 in proper order the light beam polarization is left-handed and the right-handed polarized light of periodic variation, the left-handed and right-handed polarized light of periodic variation focuses on to the sample district of surveyed sample 8 through reflective objective and transmits and passes through surveyed sample 8, after being collimated by reflective objective two 9 of measured sample 8's transmission light, focuses on grating spectrometer 12 through first 11, the beam splitter 12 will focus the light beam receives the grating in the wavelength direction, and fix on having the grating axle of the spectral beam splitter and around the displacement of the optical axis and can drive the grating displacement of freely rotating around the optical axis and the grating 13, the grating displacement of the free-axis of the grating that can be around the grating is driven under the photoelectric multiplier displacement of the wavelength is passed through the optical axis.
As shown in fig. 2, in the data collecting and processing process, the photoelastic modulation controller 501 generates a square wave signal with a period of 50kHz, the square wave signal controls the optical axis of the photoelastic modulator 5 to rotate, so that the excitation light transmitted through the photoelastic modulator is periodically changed into left-handed polarized light and right-handed polarized light, the square wave signal with the period of 50kHz simultaneously triggers the synchronous controller 17, the synchronous controller provides a timing signal to the photomultiplier and the single photon counting and collecting card 13 18 according to the actual system condition, the single photon counting and collecting card 18 calibrates the collecting timing of the photomultiplier 13, and simultaneously distinguishes the left-handed polarized light component and the right-handed polarized light component of the light signal collected by the photomultiplier 13, in the data processing process, the left-handed polarized light intensity and the right-handed polarized light intensity of a single wavelength can be obtained through single collection, and the wavelength is changed through rotating the grating spectrometer 12, so that the circular dichroism spectrum of the measured sample 8 in the whole spectrum section is obtained.
In this embodiment, the broad spectrum continuous light source 1 is preferably a deuterium-halogen lamp, and the first reflective objective 7 and the second reflective objective 9 are preferably objectives with a focal length of 200 mm.
Claims (10)
1. A micro circular dichroism spectrum detection system based on a single photon counting and collecting method is characterized by comprising a light source module, a polarizing module, an excitation module and a collecting module which are sequentially arranged in a line, and further comprising a data processing module, a micro imaging module, a synchronization module and a white light source (14);
the data processing module is respectively connected with the acquisition module and the synchronization module through data transmission lines, the synchronization module is connected with the polarizing module through the data transmission lines, the microscopic imaging module is arranged on a refraction light path of the excitation module, and the white light source (14) is arranged on a reflection light path of the excitation module.
2. The system for detecting the microscopic circular dichroism spectrum based on the single photon counting collection method according to claim 1, wherein the light source module comprises a broad spectrum continuous light source (1) and a first reflecting mirror (2) and a second reflecting mirror (3) which are sequentially arranged on the light path of the broad spectrum continuous light source.
3. The system for detecting the dichroism in microscopic circles based on the single photon counting collection method according to claim 1, wherein the polarization module comprises a polarizer (4) and a photoelastic modulator (5) which are sequentially arranged in a line, the polarizer (4) and a second reflector (3) are arranged in a line, and the synchronization module is connected with the photoelastic modulator (5) through a data transmission line.
4. The system of claim 1, wherein the excitation module comprises: the device comprises a movable beam splitting piece (6), a first reflective objective lens (7), a second reflective objective lens (9) and a movable reflector (10), wherein the movable beam splitting piece (6) and the photoelastic modulator (5) are sequentially arranged in a line, the microscopic imaging module is arranged on a refraction light path of the movable beam splitting piece (6), a white light source (14) is arranged on a reflection light path of the movable reflector (10), and a sample area used for placing a tested sample (8) is arranged between the first reflective objective lens (7) and the second reflective objective lens (9).
5. The system for detecting the microscopic circular dichroism spectrum based on the single photon counting and collecting method according to claim 1, wherein the collecting module comprises a first focusing mirror (11) and a grating light splitter (12) which are sequentially arranged in a line, and further comprises a photomultiplier tube (13), the first focusing mirror (11) and the movable reflector (10) are arranged in a line, the photomultiplier tube (13) is arranged on a reflection light path of the grating light splitter (12), and the photomultiplier tube (13) is connected with the data processing module through a data transmission line.
6. The system for detecting the microscopic circular dichroism spectrum based on the single photon counting acquisition method as claimed in claim 1, wherein the data processing module comprises a single photon counting acquisition card (18) and a data processing computer (19), and the single photon counting acquisition card (18) is respectively connected with the synchronization module, the data processing computer (19) and the photomultiplier (13) through data transmission lines.
7. The system for detecting the microscopic circular dichroism spectrum based on the single photon counting and collecting method according to claim 1, wherein the microscopic imaging module comprises a second focusing mirror (15) and a CCD camera (16) which are sequentially arranged on a refraction light path of the movable beam splitting plate (6), and further comprises an imaging display (20) connected with the CCD camera (16) through a data connecting line.
8. The system for microscopic circular dichroism spectroscopy based on a single photon counting collection method according to claim 1, wherein the synchronization module comprises a synchronization controller (17), and the synchronization controller (17) is respectively connected with the photoelastic modulator (5) and the single photon counting collection card (18) through data connection lines.
9. The system according to claim 1, wherein when performing spectral measurement on a sample to be measured, the movable beam splitting plate (6) and the movable mirror (10) in the excitation module are arranged in a line, and white light emitted from the white light source (14) sequentially passes through the movable mirror (10) and the second reflective objective (9) in the excitation module to illuminate a sample area of the sample to be measured (8), so that the sample area of the sample to be measured (8) sequentially passes through the first reflective objective (7) and the movable beam splitting plate (6) and is imaged in the microscopic imaging module.
10. The system according to claim 1, wherein when spectra of the sample are collected, the movable beam splitter (6) and the movable mirror (10) in the excitation module are arranged in a line, the broad spectrum light beam emitted from the broad spectrum continuous light source (1) in the light source module sequentially passes through the polarization module and the reflective objective lens (7) in the excitation module and transmits through the sample region of the sample (8), and the transmitted light sequentially passes through the reflective objective lens (9) and the collection module and then is collected in the data processing module for the spectra of the sample (8).
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