CN113834803A - A Multifunctional Microscopic Imaging Optical System - Google Patents
A Multifunctional Microscopic Imaging Optical System Download PDFInfo
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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Abstract
本发明公开了一种多功能显微成像光学系统，本方案系统主体以显微镜、光谱仪及激发光源模块作为三大组成，将其用于样品微纳区域成像和光谱采集工作，实现了暗场成像、明场成像、荧光成像、暗场散射光谱、荧光光谱、拉曼光谱及拉曼mapping等功能。该系统可通过光路切换和测试模块切换，无需转移样品即可实现对材料的不同模式和不同方法测试分析，获得材料的多种光谱信息；本方案涉及的多功能显微成像光学系统可避免由于样品在不同仪器间转移造成的测量误差，提高了测量结果的准确性。在生命科学，纳米技术及半导体技术等领域应用广泛市场广阔。
The invention discloses a multifunctional microscopic imaging optical system. The main body of the system is composed of a microscope, a spectrometer and an excitation light source module. It is used for sample micro-nano area imaging and spectrum acquisition, and realizes dark field imaging. , bright field imaging, fluorescence imaging, dark field scattering spectroscopy, fluorescence spectroscopy, Raman spectroscopy and Raman mapping and other functions. The system can switch between optical paths and test modules, without transferring samples, it can test and analyze different modes and methods of materials, and obtain a variety of spectral information of materials; the multi-functional microscopic imaging optical system involved in this scheme can avoid the The measurement error caused by the transfer of samples between different instruments improves the accuracy of the measurement results. It is widely used in the fields of life science, nanotechnology and semiconductor technology.
Description
Technical Field
The invention relates to the technical field of optical systems, spectral measurement technologies and optical imaging, in particular to a multifunctional microscopic imaging optical system.
Background
The light reacts with the substance, and the substance can absorb, transmit, scatter and reflect photons. The substance may also emit photons after reabsorbing them. By utilizing the optical information, people develop and develop optical detection schemes such as fluorescence spectrum and imaging, dark field scattering spectrum and imaging, bright field imaging, Raman spectrum and Raman mapping and the like based on the application of the optical information. The technologies play an important role in the fields of scientific research, analysis and detection, medical treatment, archaeological identification and the like; however, in practical application and design, the spectral imaging technology and the spectral acquisition analysis technology are relatively independent. This has also led to the microscope that uses extensively on the market now and spectrum appearance function single relatively, if need carry out multiple spectral information acquisition analysis and imaging pattern analysis to the sample in the actual operation in-process, just need test on different equipment, will cause phenomenons such as result measurement error, measurement sample waste and efficiency of software testing low like this.
Although the two technologies of spectral imaging and spectral collection are relatively independent, the light path design is based on the microscope design, and partial light paths are shared, so that the possibility of combination exists. Based on the scheme, the multifunctional microscopic imaging optical system carries multiple excitation light sources, and functions of dark field imaging, bright field imaging, fluorescence imaging, dark field scattering spectrum, fluorescence spectrum, Raman mapping and the like are realized in one set of system through successful switching of the light path and the excitation module; and a more comprehensive spectrum and imaging representation result is realized in a set of optical system, and the application fields of the micro-spectrum and the imaging technology are widened.
Disclosure of Invention
In view of the above-mentioned problems, it is an object of the present invention to provide a multifunctional microscopic imaging optical system having a multifunctional detection mode, which is convenient to switch operation and has an excellent detection effect.
In order to achieve the technical purpose, the technical scheme adopted by the invention is as follows:
a multifunctional microscopic imaging optical system, comprising: the device comprises a mercury lamp light source, a halogen lamp light source, a plurality of lasers, a spectrometer CCD, a microscopic imaging CCD, a Raman auxiliary camera, a double-optical-path conversion module, a fluorescence excitation module, a bright-field excitation module, a dark-field excitation module, a Raman excitation module, an optical-path switching module and a sample table, wherein a sample to be detected is placed on the sample table;
the double-light-path conversion module is provided with an input end and an output end, the input end of the double-light-path conversion module is connected with the mercury lamp light source and the halogen lamp light source, the double-light-path conversion module receives light output by the mercury lamp light source and the halogen lamp light source and switches and outputs light input by the mercury lamp light source or the halogen lamp light source, and the light output by the output end of the double-light-path conversion module is emitted to a sample to be detected on the sample stage;
the working wavelengths of the plurality of lasers are different and are used for providing lasers with different wavelengths, and light output by the output ends of the plurality of lasers is emitted to a sample to be detected on the sample stage;
the fluorescence excitation module, the bright field excitation module, the dark field excitation module and the Raman excitation module are arranged on a light path emitted to a sample to be detected, and the fluorescence excitation module, the bright field excitation module, the dark field excitation module and the Raman excitation module can be switched into the light path or switched out of the light path.
The light emitted to the sample to be tested is emitted to the sample to be tested and returned after transmitting through the fluorescence excitation module, the bright field excitation module, the dark field excitation module or the Raman excitation module, and is emitted to the light path switching module after transmitting through the fluorescence excitation module, the bright field excitation module, the dark field excitation module or the Raman excitation module;
in addition, the light incident to the light path switching module is guided by the light path switching module, so that the light is incident to the spectrometer CCD or the microscopic imaging CCD or is partially incident to the Raman auxiliary camera.
As a possible implementation manner, further, after the light emitted by the mercury lamp light source and the halogen lamp light source is received by the dual-light path conversion module, the light shares the same light path to output; the light output by the multiple lasers is transmitted to the sample to be tested on the sample platform by one light path in common, wherein the double-light-path conversion module and the multiple lasers transmit photons to the sample to be tested on the sample platform through the reflector units, namely, for the convenience of light path integration, the light output by the double-light-path conversion module and the multiple lasers can be adjusted through the reflector units and then transmitted to the fluorescence excitation module, the bright field excitation module, the dark field excitation module and the Raman excitation module; in addition, the light which enters the sample to be measured and returns enters the light path switching module.
As a possible implementation, further, the power of the mercury lamp light source is 130W, the power of the halogen lamp light source is 100W, and the operating voltage is 12V.
As a possible implementation, further, the mercury lamp light source guides the light output by the mercury lamp light source to the input end of the dual optical path conversion module through a cold optical fiber; by introducing the cold light fiber, the heat transfer is reduced, the service life of the mercury lamp light source bulb can reach more than 2000 hours, and the light intensity is adjustable. The 12V 100W halogen lamp and the lasers with different wave bands are provided with the high-performance optical filter and the polaroid, the multi-path laser is placed in the integrated optical path box and used in a free optical path coupling mode, and the high-performance optical filter group is fully automatically switched along with the electric turntable.
As a possible embodiment, further, the fluorescence excitation module includes an ultraviolet excitation module, a blue excitation module, and a green excitation module.
As a possible implementation manner, further, the optical path switching module includes a first reflecting mirror, a second reflecting mirror, a half-reflecting and half-transmitting mirror, a first neutral position box and a second neutral position box which can be switched into or cut out from the optical path, and the raman auxiliary camera is disposed in the direction of the reflected optical path of the first reflecting mirror and the half-reflecting and half-transmitting mirror; the second reflector and the second neutral gear box are arranged in the transmission light path direction of the transflective mirror or the first neutral gear box; the spectrometer CCD is arranged in the direction of a reflection light path of the second reflecting mirror, and the microscopic imaging CCD is arranged in the direction of a transmission light path of the second neutral gear box.
As a preferred implementation option, preferably, the light incident to the CCD of the spectrometer through the light path switching module also passes through an electric turntable, and the electric turntable is provided with a long pass filter which can be switched into or cut out from the light path.
As a preferred implementation option, preferably, a third reflecting mirror and a light splitting path are further disposed on a light path between the electric turntable and the spectrometer CCD.
As a preferred implementation choice, the operating wavelengths output by the plurality of laser light sources are 523nm, 633nm and 785nm, respectively.
As a preferred implementation option, it is preferred that the system includes a raman test mode, in the raman test mode, one of the plurality of laser light sources is used as a detection light source, and the raman excitation module, the long-pass filter on the electric turntable, and the half-reflecting and half-transmitting mirror of the optical path switching module are switched into the optical path;
after the laser with the corresponding working wavelength is output by one of the laser sources, the laser is emitted into a sample to be detected on the sample platform through the Raman excitation module, the laser returns through the sample to be detected and then is emitted into the light path switching module through the Raman excitation module again, the light path switching module switches the half-reflecting half-transmitting mirror and the second reflecting mirror into a light path, so that part of the laser is reflected into the Raman auxiliary camera, part of the laser which penetrates through the half-reflecting half-transmitting mirror is emitted into the second reflecting mirror and is reflected to the electric turntable after passing through the second reflecting mirror, the electric turntable switches the long-pass filter into the light path, and the laser which is reflected to the electric turntable penetrates through the long-pass filter and is received by the CCD spectrometer after sequentially passing through the third reflecting mirror and the light splitting light path;
in addition, when the working wavelength output by the laser light source is 523nm, the Raman frequency shift range is 50-6000cm-1When the working wavelength of the laser light source output is 785nm laser, the Raman frequency shift range is 50-3200cm-1The extinction ratio of the polarization spectrum test range of 510-800nm is more than 1000:1, and the extinction ratio of the polarization spectrum test range of 750-1800nm is more than 10000: 1; the system has high-efficiency luminous flux (more than 50 percent) and high sensitivity, the signal-to-noise ratio of the third-order Raman peak of silicon is better than 30:1 when the Raman signal of monocrystalline silicon is tested, and the fourth-order Raman peak can be observed.
Based on the structural scheme, the scheme can realize the detection work of fluorescence imaging, fluorescence sampling spectrum, bright field imaging, dark field scattering spectrum, Raman spectrum and Raman mapping.
By adopting the technical scheme, compared with the prior art, the invention has the beneficial effects that: the mercury lamp light source, the halogen lamp light source, the multiple lasers, the spectrometer CCD, the microscopic imaging CCD, the Raman auxiliary camera, the double-light-path conversion module, the fluorescence excitation module, the bright field excitation module, the dark field excitation module, the Raman excitation module, the light path switching module and the sample platform are matched, the reflector unit is used for guiding the light source to input into a light path, the formed system main body comprises a microscope, the spectrometer and the excitation light source module, and the system main body is used for sample micro-nano area imaging and spectrum acquisition, so that functions of dark field imaging, bright field imaging, fluorescence imaging, dark field scattering spectrum, fluorescence spectrum, Raman mapping and the like are realized. The system can realize the test analysis of different modes and different methods of the material without transferring samples by switching the light path and the test module, and obtain various spectral information of the material; the multifunctional microscopic imaging optical system can avoid measurement errors caused by transfer of samples among different instruments, and improves the accuracy of measurement results. The method is widely applied to the fields of life science, nanotechnology, semiconductor technology and the like, and has wide market.
Drawings
The invention will be further explained with reference to the drawings and the detailed description below:
FIG. 1 is a schematic diagram of a system according to the present embodiment of the invention;
FIG. 2 is a schematic diagram of one embodiment of the system;
FIG. 3 is a schematic view of a microscope with partial optical path switching and various excitation modules, in which a halogen lamp light source, a mercury lamp light source and a multi-laser form the light source, which is directed into the optical system via a mirror unit;
fig. 4 is an image of the multifunctional microscopic imaging optical system obtained in example 1, wherein fig. 4a is a bright field image, fig. 4b is a dark field image, fig. 4c is a 375nm fluorescence excitation image, fig. 4d is a 450nm fluorescence excitation image, and fig. 4e is a 520nm fluorescence excitation image;
FIG. 5 shows fluorescence spectra obtained in example 2;
FIG. 6 shows the Raman spectrum obtained in example 2.
Detailed Description
As shown in one of fig. 1 to 3, the present invention provides a multifunctional microscopic imaging optical system, which includes: the device comprises a mercury lamp light source 1, a halogen lamp light source 2, a plurality of lasers 4, a spectrometer CCD9, a microscopic imaging CCD, a Raman auxiliary camera 11, a double-optical-path conversion module 3, a fluorescence excitation module, a bright-field excitation module 16, a dark-field excitation module 17, a Raman excitation module 18, an optical-path switching module 12 and a sample table 19, wherein a sample 191 to be detected is placed on the sample table 19;
the double-light-path conversion module 3 is provided with an input end and an output end, the input end of the double-light-path conversion module 3 is connected with the mercury lamp light source 1 and the halogen lamp light source 2, the double-light-path conversion module 3 receives the light output by the mercury lamp light source 1 and the halogen lamp light source 2 and switches and outputs the light input by the mercury lamp light source 1 or the halogen lamp light source 2, and the light output by the output end of the double-light-path conversion module 3 is emitted to the sample 191 to be measured on the sample stage 19;
the working wavelengths of the plurality of lasers 4 are different and are used for providing lasers with different wavelengths, and light output by the output ends of the plurality of lasers 4 is emitted to a sample 191 to be detected on the sample stage 19;
the fluorescence excitation module, the bright field excitation module 16, the dark field excitation module 17 and the raman excitation module 18 are arranged on a light path emitted to the sample 191 to be measured, and the fluorescence excitation module, the bright field excitation module 16, the dark field excitation module 17 and the raman excitation module 18 can be switched into or switched out of the light path.
Wherein, the light emitted to the sample 191 to be measured is emitted to the sample 191 to be measured and returned after being transmitted through the fluorescence excitation module, the bright field excitation module 16, the dark field excitation module 17 or the raman excitation module 18, and is emitted to the light path switching module 12 after being transmitted through the fluorescence excitation module, the bright field excitation module 16, the dark field excitation module 17 or the raman excitation module 18;
in addition, the light incident on the light path switching module 12 is guided by the light path switching module 12, and is incident on the spectrometer CCD9 or the microscopic imaging CCD10 or partially incident on the raman auxiliary camera 11.
In this embodiment, as a possible implementation manner, further, the fluorescence excitation module includes an ultraviolet excitation module 13, a blue excitation module 14, and a green excitation module 15.
In this embodiment, as a preferred implementation choice, the operating wavelengths output by the plurality of laser light sources 4 are preferably 523nm, 633nm, and 785nm, respectively.
In this embodiment, as a possible implementation manner, further, after the light emitted by the mercury lamp light source 1 and the halogen lamp light source 2 is received by the dual light path conversion module 3, the light shares the same light path to output; the light output by the plurality of lasers 4 is emitted to the sample 191 to be measured on the sample stage 19 by sharing one light path, wherein the dual light path conversion module 3 and the plurality of lasers 4 transmit photons to the sample 191 to be measured on the sample stage 19 by the reflector unit 8, that is, for the convenience of light path integration, the light output by the dual light path conversion module 3 and the plurality of lasers 4 can be adjusted by the reflector unit 8 to be emitted to the fluorescence excitation module, the bright field excitation module 16, the dark field excitation module 17 and the raman excitation module 18; in addition, the light incident into the sample 191 to be measured and returning enters the optical path switching module 12.
In this embodiment, as a possible implementation manner, the power of the mercury lamp light source 2 is 130W, the power of the halogen lamp light source 2 is 100W, and the operating voltage is 12V.
In this embodiment, as a possible implementation manner, the mercury lamp light source 1 further guides the light output by the mercury lamp light source to the input end of the dual optical path conversion module 3 through a cold optical fiber.
In this embodiment, as a possible implementation manner, the optical path switching module 12 further includes a first reflecting mirror 8-1, a second reflecting mirror 8-2, a half-reflecting and half-transmitting mirror 22, a first neutral position box 23-1, and a second neutral position box 23-2 that can be switched into or cut out from the optical path, and the raman auxiliary camera 11 is disposed in the direction of the reflection optical path of the first reflecting mirror 8-1 and the half-reflecting and half-transmitting mirror 22; the second reflector 8-2 and the second neutral box 23-2 are arranged in the transmission light path direction of the half-reflecting and half-transmitting mirror 22 or the first neutral box 23-1; the spectrometer CCD9 is arranged in the direction of a reflection light path of the second reflecting mirror 8-2, and the microscopic imaging CCD10 is arranged in the direction of a transmission light path of the second neutral box 23-2.
In this embodiment, as a preferred implementation choice, preferably, the light incident to the spectrometer CCD9 through the light path switching module 12 also passes through the electric turntable 5, and the electric turntable 5 is provided with a long pass filter 6 capable of being switched into or cut out from the light path.
In the scheme, as a preferred implementation choice, preferably, a third reflecting mirror 8-3 and a light splitting light path 7 are further arranged on a light path between the electric turntable 5 and the spectrometer CCD 9.
In this embodiment, as a preferred implementation choice, it is preferred that the system includes a raman test mode, in the raman test mode, one of the plurality of laser light sources 4 is used as a detection light source, and the raman excitation module 18, the long-pass filter 6 on the electric turntable 5, and the half-reflecting and half-transmitting mirror 22 of the optical path switching module 12 are switched into the optical path.
Wherein, after the laser with corresponding working wavelength is output by one of the plurality of laser sources 4, the laser is emitted into a sample 191 to be measured on the sample platform 19 through the Raman excitation module 18, the laser is returned through the sample 191 to be measured, then is emitted into the optical path switching module 12 through the Raman excitation module 18 again, the optical path switching module 12 switches the half-reflecting and half-transmitting mirror 22 and the second reflecting mirror 8-2 into the optical path, so that part of the laser is reflected into the Raman auxiliary camera 11, part of the laser which transmits the half-reflecting and half-transmitting mirror 22 is emitted into the second reflecting mirror 8-2, and is reflected to the electric turntable 6 through the second reflecting mirror 8-2, the electric turntable 6 switches the long-pass filter 6 into the optical path, so that the laser which is reflected to the electric turntable 5 transmits the long-pass filter 6, and sequentially passes through the third reflecting mirror 8-3 and the light splitting optical path, received by the spectrometer CCD 9;
in addition, when the working wavelength output by the laser light source is 523nm, the Raman frequency shift range is 50-6000cm-1When the working wavelength of the laser light source output is 785nm laser, the Raman frequency shift range is 50-3200cm-1The extinction ratio of the polarization spectrum test range of 510-800nm is larger than 1000:1, and the extinction ratio of the polarization spectrum test range of 750-1800nm is larger than 10000: 1.
Based on the structure scheme, the scheme can realize the detection work of fluorescence imaging, fluorescence sampling spectrum, bright field imaging, dark field scattering spectrum, Raman spectrum and Raman mapping, and the corresponding work coordination mode is as follows:
fluorescence imaging and fluorescence spectroscopy: the mercury lamp light source 1 is activated to emit photons, the photons enter a microscope light path through the double-light-path conversion module 3 by adjusting the double-light-path conversion module 3, and enter a fluorescence excitation module (such as an ultraviolet excitation module 13, a blue light excitation module 14 and a green light excitation module 15) through the reflector unit 8, wherein a specific filtering excitation module (the ultraviolet excitation module 13, the blue light excitation module 14 or the green light excitation module 15) in the fluorescence excitation module can pass through a specific light wavelength; photons transmitted through the fluorescence excitation module act with a sample 191 to be detected on the sample platform 19, and photons emitted by a detected object after returning pass through a specific fluorescence excitation module, such as an ultraviolet excitation module 13, a blue light excitation module 14 or a green light excitation module 15, and then enter the optical path switching module 12; the optical path switching module 12 has a first mirror 8-1, a half-reflecting half-transmitting mirror 22, a first neutral box 23-1, a second mirror 8-2, and a second neutral box 23-2 that can switch an incoming optical path and an outgoing optical path. Photons pass through the second reflecting mirror 8-2 and no optical component is arranged on the light path by adjusting the position of the electric turntable 5, the photons enter the spectrometer CCD9 to obtain fluorescence spectrum information, and the light path switching module 12 is adjusted to enable the photons to enter the microscopic imaging CCD10 through the first blank box 23-1 and the second blank box 23-2 to obtain a fluorescence imaging pattern.
Bright field imaging: a halogen lamp 2 is selected as a light source, and the double-light-path conversion module 3 is adjusted to enable photons to enter a microscope light path and enter a scattered bright field excitation module 16 through a reflector unit 8. The photons react with a detection sample 191 on a sample stage 19, then return to enter a scattering bright field excitation module 16, pass through a first neutral box 23-1 and a second neutral box 23-2 in an optical path switching module 12, and enter a microscopic imaging CCD10 to obtain a scattering bright field imaging image.
Dark field imaging and dark field scattering spectroscopy: a halogen lamp 2 is selected as a light source, and the double-light-path conversion module 3 is adjusted to enable photons to enter a microscope light path and enter a scattering dark field excitation module 17 through a reflector unit 8. The photons react with a detection sample 191 on a sample table 19 and then enter a dark field scattering module 17, the photons are switched to a second reflecting mirror 8-2 when passing through an optical path switching module 12, the position of an electric turntable 5 is adjusted to enable no optical component to be arranged on the optical path, and the photons enter a spectrometer CCD9 to obtain dark field scattering spectrum information. To obtain a scattered dark field imaging pattern, photons switched to the first and second neutral boxes 23-1 and 23-2 directly enter the microscopic imaging CCD10 to obtain a dark field imaging pattern as the photons pass through the optical path switching module 12.
Raman spectroscopy and raman mapping: a laser light source 4 with a specific wavelength is selected and enters a microscope light path and a Raman excitation module 18 through a reflector unit 8, laser and a sample 191 to be detected on a sample table 19 are acted, and a half-reflecting and half-transmitting mirror 22 in the light path switching module 12 is adjusted to enable partial photons to enter a Raman auxiliary camera 11. By adjusting the focal plane, the laser spot is brought into the optimum focusing state in the raman assisting camera 11. The readjustment light path switching module 12 introduces photons into the light path of the spectrometer by using the second reflecting mirror 8-2, adjusts the position of the corresponding long pass filter 6 on the electric turntable 5 corresponding to the laser wavelength, and filters most of rayleigh scattered photons, so that the raman scattered photons enter the spectrometer CCD 9. When the Raman mapping function is performed, the bright field mentioned above is used for imaging, a Raman scanning area and a Raman scanning step length are selected, the sample stage 19 is provided with an electric device, and the surface distribution condition of Raman signals can be obtained by using a Raman signal acquisition light path.
Based on the technical scheme, the scheme aims to provide the multifunctional microscopic imaging spectrum system which comprises three main bodies, a microscope, a spectrometer and an excitation light source module; the optical system can realize functions of dark field imaging, bright field imaging, fluorescence imaging, dark field scattering spectrum, fluorescence spectrum, Raman mapping and the like. The following embodiments are used to further illustrate the implementation of the present invention:
example 1
On the basis of the description of one of the figures 1 to 3 and the description of figure 4, a coordination polymer formed by copper bromide and parapyrazine oxide forms a color interference pattern on the photoetching micro-nano pattern, by placing the sample 191 to be measured on the sample stage 9, turning on the halogen lamp light source 2, adjusting the dual light path conversion module 3 to make the light source enter the light path of the microscope, the light source enters the scattered bright field module 16 through the reflector unit 8, and then passes through the scattered bright field module 16 after reacting with the sample 191 to be measured, and then the first blank box 23-1 and the second blank box 23-2 in the light path switching module 12 are adjusted to cut into the light path, so that the photons enter the microscopic imaging CCD10 to obtain the bright field imaging diagram shown in FIG. 4a, and the dark field imaging diagram shown in fig. 4b can be obtained by changing the excitation module into the scattering dark field module 17 and adjusting the exposure time of the camera by keeping the light path unchanged.
During fluorescence imaging, the optical fiber mercury lamp light source 1 is used, the double-light-path conversion module 3 is adjusted to enable the light source to enter a microscope light path, the light source respectively enters the ultraviolet excitation module 13, the blue light excitation module 14 and the green light excitation module 15 through the reflector unit 8, after the light source and a sample 191 to be detected react, the light source respectively passes through the ultraviolet excitation module 13, the blue light excitation module 14 and the green light excitation module 15, photons enter the micro-imaging CCD10 through the first vacancy box 23-1 and the second vacancy box 23-2 in the light path switching module 12, and then a 375nm excited fluorescence imaging graph 4c, a 450nm excited fluorescence imaging graph 4d and a 520nm excited fluorescence imaging graph 4e can be obtained step by step.
Example 2
On the basis of one of the diagrams shown in fig. 1 to 3, with reference to fig. 5 or 6, by using plasma enhanced paper (CNFNP) prepared by using nanocellulose and silver cubic nanoparticles as raw materials and paper (CNP) prepared by using nanocellulose as a contrast, fluorescent dye molecule fluorescein and raman probe molecule thiophenol with the same concentration are printed on the two paper sheets respectively, a sample 191 to be detected is placed on a sample table 19, a light source 1 of a fiber-optic mercury lamp is turned on, a double-light-path conversion module 3 is adjusted to enable the light source to enter a microscope light path, at this time, a blue light excitation module 14 is selected as a filtering module to obtain an excitation light source of 450nm to excite a sample, after the light source and the sample act, the blue light excitation module 14 is passed through, then a reflecting mirror unit 8 in a light path switching module 12 is adjusted to enable photons to enter a spectrometer light path, an electric turntable 5 is adjusted to enable photons on the light path without a long-pass filter 6 to enter a CCD9 through a subsequent light path to obtain a fluorescence spectrum 5, comparing the two groups of data shows that the plasma enhanced fluorescence reaches 3.6 times.
Turning off other light sources, turning on a 532nm laser 4, correspondingly starting an electric rotating platform 5, namely, enabling a corresponding 532nm long-pass filter 6 to be positioned on a light path, enabling 532nm laser to enter a microscope light path through a reflector unit 8, firstly adjusting a half-reflecting half-transmitting mirror in a light path switching module 12, enabling the laser to act with a sample, enabling a part of photons to enter a Raman auxiliary camera 11 through the half-reflecting half-transmitting mirror, enabling light spots in the Raman auxiliary camera 11 to be optimally focused by adjusting a focusing surface, then adjusting a second reflector unit 8-2 in the light path switching module 12 to enable the photons to pass through the corresponding long-pass filter 6 on an electric rotating table 5, and enabling Raman scattering signals to enter a subsequent light path to obtain a Raman spectrogram 6 on a spectrometer CCD 9. Compared with data table, the surface plasma enhanced Raman spectrum is obvious.
The foregoing is directed to embodiments of the present invention, and equivalents, modifications, substitutions and variations such as will occur to those skilled in the art, which fall within the scope and spirit of the appended claims.
Claims (10)
1. A multifunctional microscopic imaging optical system is characterized in that: it includes: the device comprises a mercury lamp light source, a halogen lamp light source, a plurality of lasers, a spectrometer CCD, a microscopic imaging CCD, a Raman auxiliary camera, a double-optical-path conversion module, a fluorescence excitation module, a bright-field excitation module, a dark-field excitation module, a Raman excitation module, an optical-path switching module and a sample table, wherein a sample to be detected is placed on the sample table;
the double-light-path conversion module is provided with an input end and an output end, the input end of the double-light-path conversion module is connected with the mercury lamp light source and the halogen lamp light source, the double-light-path conversion module receives light output by the mercury lamp light source and the halogen lamp light source and switches and outputs light input by the mercury lamp light source or the halogen lamp light source, and the light output by the output end of the double-light-path conversion module is emitted to a sample to be detected on the sample stage;
the working wavelengths of the plurality of lasers are different and are used for providing lasers with different wavelengths, and light output by the output ends of the plurality of lasers is emitted to a sample to be detected on the sample stage;
the fluorescence excitation module, the bright field excitation module, the dark field excitation module and the Raman excitation module are arranged on a light path emitted to a sample to be detected, and the fluorescence excitation module, the bright field excitation module, the dark field excitation module and the Raman excitation module can be switched into the light path or cut out of the light path;
the light emitted to the sample to be tested is emitted to the sample to be tested and returned after transmitting through the fluorescence excitation module, the bright field excitation module, the dark field excitation module or the Raman excitation module, and is emitted to the light path switching module after transmitting through the fluorescence excitation module, the bright field excitation module, the dark field excitation module or the Raman excitation module;
in addition, the light incident to the light path switching module is guided by the light path switching module, so that the light is incident to the spectrometer CCD or the microscopic imaging CCD or is partially incident to the Raman auxiliary camera.
2. The multifunctional microscopic imaging optical system according to claim 1, characterized in that: the light emitted by the mercury lamp light source and the halogen lamp light source is received by the double-light-path conversion module and then output by sharing the same light path; the light output by the plurality of lasers shares one optical path and is emitted to a sample to be detected on the sample stage;
the double-optical-path conversion module and the plurality of lasers transmit photons to a sample to be detected on the sample stage through the reflector unit, the photons are transmitted to the sample to be detected, and light returned by the sample to be detected enters the optical path switching module.
3. The multifunctional microscopic imaging optical system according to claim 1, characterized in that: the power of the mercury lamp light source is 130W, the power of the halogen lamp light source is 100W, and the working voltage is 12V.
4. The multifunctional microscopic imaging optical system according to claim 1, characterized in that: the mercury lamp light source guides the light output by the mercury lamp light source into the input end of the double-light-path conversion module through the cold optical fiber.
5. The multifunctional microscopic imaging optical system according to claim 1, characterized in that: the fluorescence excitation module comprises an ultraviolet excitation module, a blue light excitation module and a green light excitation module.
6. The multifunctional microscopic imaging optical system according to claim 1, characterized in that: the optical path switching module comprises a first reflector, a second reflector, a semi-reflecting and semi-transmitting mirror, a first neutral gear box and a second neutral gear box, wherein the first reflector, the second reflector, the semi-reflecting and semi-transmitting mirror, the first neutral gear box and the second neutral gear box can be switched into an optical path or switched out of the optical path; the second reflector and the second neutral gear box are arranged in the transmission light path direction of the transflective mirror or the first neutral gear box; the spectrometer CCD is arranged in the direction of a reflection light path of the second reflecting mirror, and the microscopic imaging CCD is arranged in the direction of a transmission light path of the second neutral gear box.
7. The multifunctional microscopic imaging optical system according to claim 6, characterized in that: the light which is incident to the CCD of the spectrometer through the light path switching module also passes through the electric turntable, and the electric turntable is provided with a long-pass filter which can be switched into the light path or cut out from the light path.
8. The multifunctional microscopic imaging optical system according to claim 7, characterized in that: and a third reflector and a light splitting light path are further arranged on the light path between the electric turntable and the spectrometer CCD.
9. The multifunctional microscopic imaging optical system according to claim 8, characterized in that: the working wavelengths output by the laser light sources are 523nm, 633nm and 785nm respectively.
10. The multifunctional microscopic imaging optical system according to claim 9, characterized in that: the system comprises a Raman test mode, wherein in the Raman test mode, one of a plurality of laser light sources is used as a detection light source, and the Raman excitation module, a long-pass filter on the electric turntable and a half-reflecting and half-transmitting mirror of the light path switching module are switched to enter a light path;
after the laser with the corresponding working wavelength is output by one of the laser sources, the laser is emitted into a sample to be detected on the sample platform through the Raman excitation module, the laser returns through the sample to be detected and then is emitted into the light path switching module through the Raman excitation module again, the light path switching module switches the half-reflecting half-transmitting mirror and the second reflecting mirror into a light path, so that part of the laser is reflected into the Raman auxiliary camera, part of the laser which penetrates through the half-reflecting half-transmitting mirror is emitted into the second reflecting mirror and is reflected to the electric turntable after passing through the second reflecting mirror, the electric turntable switches the long-pass filter into the light path, and the laser which is reflected to the electric turntable penetrates through the long-pass filter and is received by the CCD spectrometer after sequentially passing through the third reflecting mirror and the light splitting light path;
in addition, when the working wavelength output by the laser light source is 523nm, the Raman frequency shift range is 50-6000cm-1When the working wavelength of the laser light source output is 785nm laser, the Raman frequency shift range is 50-3200cm-1The extinction ratio of the polarization spectrum test range of 510-800nm is larger than 1000:1, and the extinction ratio of the polarization spectrum test range of 750-1800nm is larger than 10000: 1.
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